ALBERT

Description:    The major structural and enzymatically active protein in spicules from siliceous sponges, e.g., for Suberites domuncula studied here, is silicatein. Silicatein has been established to be the key enzyme that catalyzes the formation of biosilica, a polymer that represents the inorganic scaffold for the spicule. In the present study, it is shown, by application of high-resolution transmission and scanning transmission electron microscopy that, during the initial phase of spicule synthesis, nanofibrils with a diameter of around 10 nm are formed that comprise bundles of between 10 and 20 nanofibrils. In intracellular vacuoles, silicasomes, the nanofibrils form polar structures with a pointed tip and a blunt end. In a time-dependent manner, these nanofibrillar bundles become embedded into a Si-rich matrix, indicative for the formation of biosilica via silicatein molecules that form the nanofibrils. These biosilicified nanofibrillar bundles become extruded from the intracellular space, where they are located in the silicasomes, to the extracellular environment by an evagination process, during which a cellular protrusion forms the axial canal in the growing spicule. The nanofibrillar bundles condense and progressively form the axial filament that becomes localized in the extracellular space. It is concluded that the silicatein-composing nanofibrils act not only as enzymatic silica bio-condensing platforms but also as a structure-giving guidance for the growing spicule. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1519-0 Authors Werner E. G. Müller, ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Duesbergweg 6, 55128 Mainz, Germany Enrico Mugnaioli, Institute of Physical Chemistry, Johannes Gutenberg University, Welderweg 11, 55099 Mainz, Germany Heinz C. Schröder, ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Duesbergweg 6, 55128 Mainz, Germany Ute Schloßmacher, ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Duesbergweg 6, 55128 Mainz, Germany Marco Giovine, Università degli Studi di Genova, Facoltà di Scienze Matematiche, Fisiche e Naturali, DISTAV-University of Genova, Corso Europa 26, 16132 Genova, Italy Ute Kolb, Institute of Physical Chemistry, Johannes Gutenberg University, Welderweg 11, 55099 Mainz, Germany Xiaohong Wang, ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Duesbergweg 6, 55128 Mainz, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The distribution of cochlin and its associated basement membrane proteins (collagen IV, collagen II, laminin-β2, and nidogen-1) were evaluated in the vestibular endorgans of subjects with Meniere’s disease and compared with normal specimens. Cochlin mRNA expression in vestibular endorgans from Meniere’s disease specimens was also investigated. Specimens were obtained from patients who had Meniere’s disease and who were undergoing ablative labyrinthectomy. Control specimens were obtained both from autopsy specimens with documented normal audiovestibular function and from patients undergoing labyrinthectomy for acoustic neuroma excision. In the normal control specimens, cochlin immunoreactivity was found evenly distributed in the stroma of the cristae ampullaris and maculae of the utricle. In Meniere’s specimens, cochlin immunoreactivity was markedly increased; this was associated with an increase in cochlin mRNA expression as shown by real-time reverse transcription with the polymerase chain reaction. Collagen IV and laminin-β2 immunoreactivity was significantly decreased in Meniere’s specimens. Nidogen-1 and collagen II immunoreactivity was unchanged in Meniere’s specimens when compared with normal samples. Cochlin upregulation has been implicated in the hereditary audiovestibulopathy, DFNA9. The increased expression of cochlin and decreased expression of collagen IV and laminin in Meniere’s disease are suggestive that the overexpression of cochlin contributes to the dysfunctional inner ear homeostasis seen in this disease. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1481-x Authors Audrey P. Calzada, Department of Head and Neck Surgery, UCLA School of Medicine David Geffen, 10833 Le Conte Avenue, Los Angeles, CA 90095, USA Ivan A. Lopez, Department of Head and Neck Surgery, UCLA School of Medicine David Geffen, 10833 Le Conte Avenue, Los Angeles, CA 90095, USA Luis Beltran Parrazal, Center of Cerebral Investigation, Veracruzana University, Veracruz, Mexico Akira Ishiyama, Department of Head and Neck Surgery, UCLA School of Medicine David Geffen, 10833 Le Conte Avenue, Los Angeles, CA 90095, USA Gail Ishiyama, Neurology Department, UCLA School of Medicine David Geffen, 10833 Le Conte Avenue, Los Angeles, CA 90095, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Glutamine synthetase (GS) catalyzes the de novo synthesis of glutamine, an amino acid that has been shown to influence sperm motility in mammals. To date, no information is available about GS content in human sperm. In this study, we have characterized the presence and cellular location of GS in fresh human normozoospermic samples. We have detected a single band corresponding to GS by Western blot. Confocal analysis has revealed GS immunoreactivity in the post-acrosomal head region. Moreover, double-labeling experiments with either F-actin or calicin have demonstrated GS confinement in the post-acrosomal region of the perinuclear theca. These data have been validated by a post-embedding ultra-structural study. The presence of GS in the post-acrosomal region of the perinuclear theca suggests that human sperm can carry out in glutamine synthesis. Content Type Journal Article Category Short Communication Pages 1-5 DOI 10.1007/s00441-012-1465-x Authors Maria Manuela Francou, Departamento de Biotecnología, Universidad de Alicante, Ctra. San Vicente-Alicante S/N, 03080 Alicante, Spain José R. Hombrebueno, Departamento de Biotecnología, Universidad de Alicante, Ctra. San Vicente-Alicante S/N, 03080 Alicante, Spain Joaquín De Juan, Departamento de Biotecnología, Universidad de Alicante, Ctra. San Vicente-Alicante S/N, 03080 Alicante, Spain Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The dynactin p150 Glued subunit, encoded by the gene DCTN1 , is part of the dynein-dynactin motor protein complex responsible for retrograde axonal transport in motor neurons. The p150 subunit is a candidate gene for neurodegenerative diseases, in particular motor neuron and extrapyramidal diseases. Tubulin-binding cofactors are believed to be involved in tubulin biogenesis and degradation and therefore to contribute to microtubule functional diversity and regulation. A yeast-two-hybrid screen for putative interacting proteins of dynactin p150 Glued has revealed tubulin-folding cofactor B (TBCB). We analyzed the interaction of these proteins and investigated the impact of this complex on the microtubule network in cell lines and primary hippocampal neurons in vitro. We especially concentrated on neuronal morphology and synaptogenesis. Overexpression of both proteins or depletion of TBCB alone does not alter the microtubule network and/or neuronal morphology. The demonstration of the interaction of the transport molecule dynactin and the tubulin-regulating factor TBCB is thought to have an impact on several cellular mechanisms. TBCB expression levels have been found to have only a subtle influence on the microtubule network and neuronal morphology. However, overexpression of TBCB leads to the decreased localization of p150 to the microtubule network that might result in a functional modulation of this protein complex. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1463-z Authors Georges F. Kuh, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Marianne Stockmann, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Marie Meyer-Ohlendorf, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Leonhard Linta, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Christian Proepper, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Albert C. Ludolph, Department of Neurology, Ulm University, Ulm, Germany Juergen Bockmann, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Tobias M. Boeckers, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Stefan Liebau, Institute for Anatomy & Cell Biology, Ulm University, Albert-Einstein Allee 11, 89081 Ulm, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Pulmonary intravascular macrophages (PIMs) are present in species such as cattle, sheep and horse and promote acute lung inflammation (ALI). Rabbits are often used as a model of ALI but there is controversy about the presence of PIMs in these species. Rabbits were treated with 10 mg/kg of gadolinium chloride intravenously (GC; n  = 6) or saline ( n  = 6) followed by euthanasia at 48 h post-treatment to determine the presence of PIMs. In a subsequent study, rabbits were pre-treated with GC or 0.9 % saline followed by 100 μg/kg of E . coli lipopolysaccharide intravenously 48 h later. Rabbits were euthanized 24 h post-LPS treatment. Light and electron microscopy showed that PIMs attached to the capillary endothelium and were positive for RAM-11 anti-macrophage antibody. While GC treatment induced apoptotic PIMs, there was no difference in the PIM number between control and GC-treated rabbits. Rabbits administered with LPS were 3.5 times more likely to die before the end of the 24-h period than those pre-treated with GC. Lung heterophil accumulation and IL-1β, TNFα and IL-6 mRNA expression were significantly higher in rabbits administered with LPS compared to those administered with GC before the LPS injection. PIMs from the LPS-treated rabbits were positive for TNFα. Lung, BAL and serum IL-8 and MCP-1 expression was not different between LPS rabbits with or without pre-treatment with GC. We conclude that rabbit lungs contain PIMs and that their depletion reduces endotoxin-induced lung inflammation. The presence of PIMs in rabbit lungs may need to be considered while using rabbit to model acute lung injury. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1509-2 Authors Tanya Duke-Novakovski, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4, Canada Sarabjeet Singh-Suri, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4, Canada Osamu Kajikawa, Medical Research Service of the VA Puget Sound Health Center, Seattle, WA, USA Sarah Caldwell, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4, Canada Chandarshekhar Charavaryamath, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4, Canada Baljit Singh, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4, Canada Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    In the atherosclerotic plaque, smooth muscle cells (SMC) acquire an inflammatory phenotype. Resistin and fractalkine (CX3CL1) are found in human atheroma and not in normal arteries. CX3CL1 and CX3CR1 are predominately associated with SMC. We have questioned whether resistin has a role in the expression of CX3CL1 and CX3CR1 in SMC thus contributing to the pro-inflammatory status of these cells. Cultured human aortic SMC were stimulated with 100 ng/ml resistin for 4, 6, 12, and 24 h, and then CX3CL1 and CX3CR1 expression was assessed by quantitative reverse transcription with the polymerase chain reaction and Western blot. We found that resistin up-regulated CX3CL1 and CX3CR1 in SMC and induced the phosphorylation of p38MAPK and STAT3. Inhibitors of p38MAPK, JAK-STAT, NF-kB, and AP-1 significantly reduced CX3CL1 and CX3CR1 expression. Knockdown of STAT1 and STAT3 with decoy oligodeoxinucleotides and the silencing of p65 and cjun with short interfering RNA decreased CX3CL1 and CX3CR1 expression. Anti-TLR4 antibody and pertussis toxin also reduced CX3CL1 and CX3CR1 protein expression. xCELLigence experiments revealed that resistin probably uses Gi-proteins for its effect on SMC. The CX3CL1 induced by resistin exhibited a chemotactic effect on monocyte transmigration. Thus, (1) resistin contributes to the pro-inflammatory state of SMC by the up-regulation of CX3CL1 and CX3CR1 expression via a mechanism involving NF-kB, AP-1, and STAT1/3 transcription factors, (2) resistin employs TLR4 and Gi-protein signaling for its effect on SMC, (3) CX3CL1 induced by resistin is functional in monocyte chemotaxis. The data reveal new mechanisms by which resistin promotes the inflammatory phenotype of SMC. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1510-9 Authors Ana-Maria Gan, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Elena Dragomir Butoi, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Adrian Manea, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Viorel Simion, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Daniela Stan, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Monica-Madalina Parvulescu, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Manuela Calin, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Ileana Manduteanu, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Maya Simionescu, Institute of Cellular Biology and Pathology “Nicolae Simionescu”, 8 B.P. Hasdeu Street, PO Box 35-14, Bucharest, Romania Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Receptor for advanced glycation end products (RAGE) is a pattern recognition receptor that binds a variety of pro-inflammatory ligands. Its soluble form, sRAGE, can compete for ligand binding and thereby have an anti-inflammatory effect. We have recently reported that sRAGE also exerts pro-inflammatory and chemotactic properties suggesting a dual role for sRAGE in immune modulation. Our present aim was to analyse the immunomodulatory properties of sRAGE in vivo with respect to acquired immunity. Naive mice were treated intra-peritoneally with sRAGE and cells from peritoneal lavage, spleens and bone marrow were examined. Mice treated with sRAGE displayed an increased leucocyte count in the peritoneal cavity, enlarged spleens and increased cellularity compared with vehicle-treated animals. Furthermore, sRAGE-treated mice had a significantly increased frequency and number of CD19 + B cells in spleen and a reduced frequency of CD19 + B cells in bone marrow compared with controls. Functionally, splenocytes from sRAGE-treated mice showed elevated IgG production and up to a four-fold increased IgM secretion compared with control animals and produced significantly higher levels of interleukin-10, interferon-γ and interleukin-6 in response to lipopolysaccharide stimulation. Our results suggest that sRAGE has immunomodulatory properties, since intra-peritoneal administration of sRAGE into healthy mice leads to rearrangements in cellular composition in the bone marrow and spleen. Moreover, the administration of sRAGE directs B cells into the spleen and towards differentiation. Our novel findings indicate that sRAGE exerts an effect on the cells of adaptive immunity. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1508-3 Authors Mikael Brisslert, Department of Rheumatology and Inflammation Research, The Sahlgrenska Academy at the University of Gothenburg, Guldhedsgatan 10A, 41346 Gothenburg, Sweden Sylvie Amu, Department of Rheumatology and Inflammation Research, The Sahlgrenska Academy at the University of Gothenburg, Guldhedsgatan 10A, 41346 Gothenburg, Sweden Rille Pullerits, Department of Rheumatology and Inflammation Research, The Sahlgrenska Academy at the University of Gothenburg, Guldhedsgatan 10A, 41346 Gothenburg, Sweden Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    This study evaluated the levels of bone morphogenetic protein receptors BMPRIB and BMPRII mRNA in goat follicles and the effects of bone morphogenetic protein-15 (BMP-15) on the in vitro development of cultured preantral follicles. Real-time polymerase chain reaction (PCR) was used to analyze the levels of BMPRIB and BMPRII mRNA in caprine preantral follicles and in small and large antral follicles. Preantral follicles (≥150 μm) were also isolated from goat ovaries and cultured for 18 days in α-MEM + supplemented with or without BMP-15 (10, 50, or 100 ng/ml). At the end of culture, some follicles were fixed for ultrastructural evaluation. Real-time PCR showed a reduction in BMPRII mRNA levels from the primary to secondary follicles. Higher levels of BMPRIB mRNA were observed in granulosa/theca cells from large antral follicles compared with small antral follicles. Moreover, BMPRII mRNA was expressed to a greater extent in cumulus–oocyte complexes from large antral follicles than in their respective granulosa/theca cells. In culture, 50 ng/ml BMP-15 positively influenced antral cavity formation and follicle growth after 18 days and also maintained follicular integrity. Thus, BMPRIB and BMPRII mRNAs are present in all follicular categories. BMP-15 (50 ng/ml) stimulates growth, antrum formation and the ultrastructural integrity of isolated caprine preantral follicles after 18 days of culture. Content Type Journal Article Category Regular Article Pages 225-238 DOI 10.1007/s00441-012-1361-4 Authors Isadora Machado T. Lima, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Ivina R. Brito, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Rafael Rossetto, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Ana Beatriz G. Duarte, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Giovanna Q. Rodrigues, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Márcia Viviane A. Saraiva, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil José Jackson N. Costa, Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, CE, Brazil Mariana Aragão M. Donato, Ultrastructure Laboratory, Aggeu Magalhães Research Center (FIOCRUZ) and Center for Strategic Technologies of the Northeast (CETENE), Recife, PE, Brazil Christina A. Peixoto, Ultrastructure Laboratory, Aggeu Magalhães Research Center (FIOCRUZ) and Center for Strategic Technologies of the Northeast (CETENE), Recife, PE, Brazil José Roberto V. Silva, Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, CE, Brazil José Ricardo de Figueiredo, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Ana Paula R. Rodrigues, Faculty of Veterinary, Laboratory of Manipulation of Oocyte and Preantral Follicles (LAMOFOPA), State University of Ceará, Fortaleza, CE, Brazil Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Recent studies have established the involvement of nasal-associated lymphoid tissues, mainly the pharyngeal tonsil, in prion pathogenesis. However, the mechanisms of the associated neuroinvasion are still debated. To determine potential sites for prion neuroinvasion inside the ovine pharyngeal tonsil, the topography of heavy (200 kDa) and light (70 kDa) neurofilaments and of glial fibrillar acidic protein has been semi-quantitatively analysed inside the various compartments of the tonsil. The results show that the most innervated areas are the interfollicular area and the connective tissue located beneath the respiratory epithelium. The existence of rare synapses between follicular dendritic cells and nerve fibres inside the germinal centre indicates that this mechanism of neuroinvasion is possible but, since germinal centres of lymphoid follicles are poorly innervated, other routes of neuroinvasion are likely. The host PRNP genotype does not influence the pattern of innervation in these various tonsil compartments, unlike ageing during which an increase of nerve endings occurs in a zone of high trafficking cells beneath the respiratory epithelium. A minimal age-related increase of innervation inside the lymphoid follicles has also been observed. An increase in nerve fibre density around the lymphoid follicles, in an area rich in mobile cells such as macrophages and dendritic cells capable of capturing and conveying pathogen prion protein (PrPd), might ensure more efficient infectivity, not in the early phase but in the advanced phase of lymphoinvasion after the amplification of PrPd; alternatively, this area might even act as a direct site of entry during neuroinvasion. Content Type Journal Article Category Regular Article Pages 167-176 DOI 10.1007/s00441-012-1376-x Authors Vinciane Toppets, Department of Morphology and Pathology, Laboratory of Animal Histology and Embryology, Faculty of Veterinary Medicine, University of Liege, Boulevard de Colonster, 20 Bat. B43a, 4000 Liege, Belgium Joelle Piret, Department of Morphology and Pathology, Laboratory of Animal Histology and Embryology, Faculty of Veterinary Medicine, University of Liege, Boulevard de Colonster, 20 Bat. B43a, 4000 Liege, Belgium Nathalie Kirschvink, Animal Physiology, Veterinary Department, Faculty of Sciences, University of Namur FUNDP, Namur, Belgium Frederic Lantier, Laboratory of Infectious Pathology and Immunology, INRA, Nouzilly, France Isabelle Lantier, Laboratory of Infectious Pathology and Immunology, INRA, Nouzilly, France Patricia Berthon, Laboratory of Infectious Pathology and Immunology, INRA, Nouzilly, France Georges Daube, DNA-Vision Agrifood, Faculty of Veterinary Medicine, University of Liege, Liege, Belgium Laurent Massart, Department of Animal productions, Biostatistic and Bioinformatic Applied to Veterinary Sciences Unit, Faculty of Veterinary Medicine, University of Liege, Liege, Belgium Luc Grobet, Department of Morphology and Pathology, Laboratory of Animal Histology and Embryology, Faculty of Veterinary Medicine, University of Liege, Boulevard de Colonster, 20 Bat. B43a, 4000 Liege, Belgium Nadine Antoine, Department of Morphology and Pathology, Laboratory of Animal Histology and Embryology, Faculty of Veterinary Medicine, University of Liege, Boulevard de Colonster, 20 Bat. B43a, 4000 Liege, Belgium Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    From its inception, tissue engineering has had three tenets: cells, biomaterial scaffolds and signaling molecules. Among the triad, cells are the center piece, because cells are the building blocks of tissues. For decades, cell therapies have focused on the procurement, manipulation and delivery of healthy cells for the treatment of diseases or trauma. Given the complexity and potential high cost of cell delivery, there is recent and surging interest to orchestrate endogenous cells for tissue regeneration. Biomaterial scaffolds are vital for many but not all, tissue-engineering applications and serve to accommodate or promote multiple cellular functions. Signaling molecules can be produced by transplanted cells or endogenous cells, or delivered specifically to regulate cell functions. This review highlights recent work in tissue engineering and cell therapies, with a focus on harnessing the capacity of endogenous cells as an alternative or adjunctive approach for tissue regeneration. Content Type Journal Article Category Review Pages 665-676 DOI 10.1007/s00441-012-1339-2 Authors Hemin Nie, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Chang Hun Lee, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Jiali Tan, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Chuanyong Lu, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Avital Mendelson, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Mo Chen, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Mildred C. Embree, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Kimi Kong, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Bhranti Shah, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Shuang Wang, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Shoko Cho, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Jeremy J. Mao, Tissue Engineering and Regenerative Medicine Laboratory (TERML), Columbia University Medical Center, 630 W. 168 St. – PH7E, New York, NY 10032, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 347 Journal Issue Volume 347, Number 3

Description:    Sensory input from the periphery to the brain can be severely compromised or completely abolished after an injury to the spinal cord. Evidence from animal models suggests that endogenous repair processes in the spinal cord mediate extensive sprouting and that this might be further attenuated by targeted therapeutic interventions. However, the extent to which sprouting can contribute to spontaneous recovery after human spinal cord injury (SCI) remains largely unknown, in part because few measurement tools are available in order to non-invasively detect subtle changes in neurophysiology. The proposed application of segmental sensory evoked potentials (e.g., dermatomal contact heat evoked potentials and somatosensory evoked potentials) to assess conduction in ascending pathways (i.e., spinothalamic and dorsal column, respectively) differs from conventional approaches in that individual spinal segments adjacent to the level of lesion are examined. The adoption of these approaches into clinical research might provide improved resolution for measuring changes in sensory impairments and might determine the extent by which spontaneous recovery after SCI is mediated by similar endogenous repair mechanisms in humans as in animal models. Content Type Journal Article Category Review Pages 1-8 DOI 10.1007/s00441-012-1373-0 Authors Jenny Haefeli, Spinal Cord Injury Center, University Hospital Balgrist, University of Zurich, Forchstrasse 340, 8008 Zürich, Switzerland Armin Curt, Spinal Cord Injury Center, University Hospital Balgrist, University of Zurich, Forchstrasse 340, 8008 Zürich, Switzerland Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Human periodontal ligament stem cells (PDLSCs) from extracted third molar teeth are a type of adult stem cell originating from dental tissue. PDLSCs are known to have a self-renewal capacity and multi-lineage differentiation potential. Vascular endothelial growth factor (VEGF), an angiogenic/vasculogenic factor, has been shown to stimulate endothelial cell mitogenesis and cell migration. Another growth factor, fibroblast growth factor-2 (FGF-2), a mitogenic factor, enhances osteogenesis in mesenchymal stem cells (MSCs). This study examines the effects of VEGF and FGF-2 on PDLSCs in vitro and in vivo compared with those on bone marrow stem cells (BMSCs) as a positive control. Treatment of PDLSCs with VEGF increases the accumulation of calcium nodules, alkaline phosphatase (ALP) activity and the formation of hard tissue and up-regulates the mRNA level of runt-related transcription factor 2 ( Runx2 ). In contrast, FGF-2 enhances the proliferation of PDLSCs in vitro in cell culture, where it significantly decreases calcium accumulation and ALP activity and down-regulates the expression of osteogenic gene markers (i.e., Runx2 , ALP , type I collagen ) involved in osteogenic induction. We have also transplanted PDLSCs with hydroxyapatite/tricalcium phosphate particles (HA/TCP) as carriers for each factor (VEGF, FGF-2) into nude mice and, after 8 weeks, observed the in vivo formation of hard tissue at the dorsal surface. Based on our results, we suggest that VEGF has positive effects on odonto-/osteogenic differentiation in vitro and on the formation of mineralized structure in vivo. FGF-2 might be a powerful promoter of the proliferation of progenitor cells in hard tissue regeneration but exogenous FGF-2 might inhibit terminal differentiation. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1392-x Authors Joo-Hee Lee, Biotooth Engineering Lab, Department of Oral and Maxillofacial Surgery, School of Dentistry, BK21, Craniomaxillofacial Life Science, Dental Research Institute, Seoul National University, Seoul, Republic of Korea Soyoun Um, Biotooth Engineering Lab, Department of Oral and Maxillofacial Surgery, School of Dentistry, BK21, Craniomaxillofacial Life Science, Dental Research Institute, Seoul National University, Seoul, Republic of Korea Jun-Hyeog Jang, Department of Biochemistry, School of Medicine, Inha University, Incheon, Republic of Korea Byoung Moo Seo, Biotooth Engineering Lab, Department of Oral and Maxillofacial Surgery, School of Dentistry, BK21, Craniomaxillofacial Life Science, Dental Research Institute, Seoul National University, Seoul, Republic of Korea Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Phosphorylated sperm proteins are crucial for sperm maturation and capacitation as a priori to their fertilization with eggs. In the freshwater prawn, Macrobrachium rosenbergii , a male reproduction-related protein (Mar-Mrr) was known to be expressed only in the spermatic ducts as a protein with putative phosphorylation and may be involved in sperm capacitation in this species. We investigated further the temporal and spatial expression of the Mar-Mrr gene using RT-PCR and in situ hybridization and the characteristics and fate of the protein using immunblotting and immunocytochemistry. The Mar-Mrr gene was first expressed in 4-week-old post larvae and the protein was produced in epithelial cells lining the spermatic ducts, at the highest level in the proximal region and decreased in the middle and distal parts. The native protein had a MW of 17 kDa and a high degree of serine/threonine phosphorylation. It was transferred from the epithelial cells to become a major protein at the anterior region of the sperm. We suggest that it is involved in sperm capacitation and fertilization in this open thelycal species and this is being investigated. Content Type Journal Article Category Regular Article Pages 1-15 DOI 10.1007/s00441-012-1380-1 Authors Ittipon Phoungpetchara, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Yotsawan Tinikul, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Jaruwan Poljaroen, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Narin Changklungmoa, Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok, 10400 Thailand Tanapan Siangcham, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Morakot Sroyraya, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Charoonroj Chotwiwatthanakun, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Rapeepun Vanichviriyakit, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Peter J. Hanna, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Prasert Sobhon, Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400 Thailand Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Synchronization of dominant follicle development and control of ovulation/oocyte retrieval are commonly used assisted reproductive technologies in both cattle and humans. The final maturation of the dominant follicle is intimately tied to the final maturation of the oocyte, preovulatory secretion of estradiol, preparation of follicular cells for luteinization, postovulatory secretion of progesterone and endocrine control of the oviductal and uterine environment for gamete and embryo development. The physiological maturity of a dominant/ovulatory follicle can affect the establishment and maintenance of pregnancy. Premature induction of the ovulatory process can reduce pregnancy rates and increase late embryonic/fetal mortality in cattle, which is likely mediated through inadequate oocyte competence and a compromised maternal environment. Oocyte competence increases with follicular maturity and is dependent upon acquisition of a complete complement of mRNA transcripts and establishment of the appropriate epigenetic marking of the oocyte genome before the preovulatory gonadotropin surge. Preovulatory secretion of estradiol is a reflection of follicular maturity and affects the oocyte, follicular cells, oviduct and uterus. The corpus luteum is a continuation of follicular maturation and rate of progesterone secretion following ovulation is linked to fertility. Advancements in our understanding of how the follicular microenvironment affects pregnancy establishment and maintenance will improve the efficiency of assisted reproductive technologies in all species. The purpose of this review is to discuss how follicular microenvironment, oocyte competence, preovulatory secretion of estradiol and postovulatory secretion of progesterone can affect pregnancy establishment and embryo/fetal survival, with an emphasis on cattle. Content Type Journal Article Category Review Pages 1-16 DOI 10.1007/s00441-012-1386-8 Authors Ky G. Pohler, Division of Animal Sciences, University of Missouri, Columbia, MO 65211, USA Thomas W. Geary, Fort Keogh Livestock and Range Research Laboratory, USDA-ARS, Miles City, MT 59301, USA Jacqueline A. Atkins, Division of Animal Sciences, University of Missouri, Columbia, MO 65211, USA George A. Perry, Department of Animal and Range Sciences, South Dakota State University, Brookings, SD 57007, USA Emma M. Jinks, Division of Animal Sciences, University of Missouri, Columbia, MO 65211, USA Michael F. Smith, Division of Animal Sciences, University of Missouri, Columbia, MO 65211, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Our aim was to investigate the in vivo gene expression pattern of the Guillain-Barre syndrome (GBS) with DNA microarrays and bioinformatics tools. Oral-infusion model animals mimicking human infection of GBS were analyzed. Tissue samples and body fluids were collected to perform antibody tests and biopsy assays. Gene-expression microarray was conducted with nerve tissues and GBS-related genes were elucidated via bioinformatics tools. Model animals showed typical symptoms of GBS in that mild demyelination was shown by cerebellar white matter and by lumbar enlargement of model animals. Then, 81.25% of the model animals were positive with GM1-IgG antibodies by ELISA. In the microarray analysis, 1,261 genes were identified with statistically different expression ( P  〈 0.05), 21 of which were associated with gene function analysis, gene pathway identification, signal transduction and co-expression network construction. Furthermore, quantitative PCR was used to characterize the gene expression level. We found that genes of HPRT1, PKC and PPARGC-1 were in the core of the network, while the expression of PPARGC-1, SUS2DD and AMPKA2 were significantly inhibited. A total of 21 genes were found to be actively involved in the process of protein transportation, transcriptional regulation, antigen identification and cell cycle regulation during the GBS infection period. The co-expression network indicated an important association between GBS and the 21 genes, especially the down-regulated ones. In conclusion, we demonstrated that GBS-affected hosts had a specific gene expression profile, which may guide the direction of GBS research and therapy. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1382-z Authors Feng Xue, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Dexin Zeng, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Fei Xu, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Rui Zhang, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Yang Chen, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Yuefei Yang, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 China Xiaobo Wang, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 China Zhenzhong Li, Department of Neurology of the Second Hospital of Hebei Medical University, Shijiazhuang, 050000 China Xiaorong Zhang, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009 China Yuan Jiang, APFIC, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Important advances in the development of smart biodegradable implants for axonal regeneration after spinal cord injury have recently been reported. These advances are evaluated in this review with special emphasis on the regeneration of the corticospinal tract. The corticospinal tract is often considered the ultimate challenge in demonstrating whether a repair strategy has been successful in the regeneration of the injured mammalian spinal cord. The extensive know-how of factors and cells involved in the development of the corticospinal tract, and the advances made in material science and tissue engineering technology, have provided the foundations for the optimization of the biomatrices needed for repair. Based on the findings summarized in this review, the future development of smart biodegradable bridges for CST regrowth and regeneration in the injured spinal cord is discussed. Content Type Journal Article Category Review Pages 1-21 DOI 10.1007/s00441-012-1352-5 Authors Elbert A. J. Joosten, Department of Anesthesiology, Pain Management and Research Center, Maastricht University Medical Hospital, Maastricht, The Netherlands Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Orexins (OxA and OxB) and their receptors (Ox1R and Ox2R), originally detected in the hypothalamus, have also been localized in multiple cerebral areas and peripheral organs. Thus, in addition to their central function in the regulation of food intake, arterial blood pressure, heart rate, sleep/wake cycle, sexual behaviour, arousal, and hypothalamic/hypophyseal axis, these neuropeptides may exert a local action in various peripheral organs and tissues. Emerging evidence suggests a main role of OxA and its highly specific receptor Ox1R in the male genital tract of mammals. We previously demonstrated OxA localization in Sertoli cells and spermatids of rat testis. Here, we show positive stainings of Ox1R in developing spermatocytes, and spermatids of rat testis by immunohistochemistry. The expression of Ox1R mRNA and the protein in the tissue was also established by reverse-transcription polymerase chain reaction and Western blotting respectively. The addition of OxA to fresh testis slices significantly increased testosterone (T) secretion which, conversely, was inhibited by Mullerian inhibiting substance (MIS). The sequential treatment of testis samples with the two substances highlighted an antagonizing activity of OxA versus MIS in regulating T levels. Furthermore, the stimulating effect on T production by OxA was prevented by the addition of the selective Ox1R inhibitor SB-408124. Overall, our findings suggest that locally secreted OxA interacting with Ox1R activates signals which antagonize MIS action in the control of T levels in mammalian testis. Content Type Journal Article Category Short Communication Pages 1-7 DOI 10.1007/s00441-012-1394-8 Authors Loredana Assisi, Department of Biological Sciences, University of Naples Federico II, Via Mezzocannone 8, 80134 Naples, Italy Simona Tafuri, Department of Biological Structures, Functions and Technologies, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy Giovanna Liguori, Department of Biological Structures, Functions and Technologies, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy Salvatore Paino, Department of Animal Production Science, University of Basilicata, Viale dell’Ateneo Lucano 10, 85100 Potenza, Italy Luigi Michele Pavone, Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Via S. Pansini 5, 80131 Naples, Italy Norma Staiano, Department of Biological Structures, Functions and Technologies, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy Alfredo Vittoria, Department of Biological Structures, Functions and Technologies, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Among arthropod pests, mites are responsible for considerable damage to crops, humans and other animals. However, detailed physiological data on these organisms remain sparse, mainly because of their small size but possibly also because of their extreme diversity. Focusing on intestinal proteases, we draw together information from three distinct mite species that all feed on skin but have separately adapted to a free-living, a strictly ecto-parasitic and a parasitic lifestyle. A wide range of studies involving immunohistology, molecular biology, X-ray crystallography and enzyme biochemistry of mite gut proteases suggests that these creatures have diverged considerably as house dust mites, sheep scab mites and scabies mites. Each species has evolved a particular variation of a presumably ancestral repertoire of digestive enzymes that have become specifically adapted to their individual environmental requirements. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-012-1369-9 Authors Deborah C. Holt, Menzies School of Health Research, Charles Darwin University, PO Box 41096, Casuarina, NT 0811, Australia Stewart T. G. Burgess, Parasitology Division, Moredun Research Institute, Pentlands Science Park, Bush Loan, Edinburgh, EH26 0PZ Midlothian, Scotland, UK Simone L. Reynolds, Infectious Diseases Program, Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Brisbane, QLD 4029, Australia Wajahat Mahmood, Menzies School of Health Research, Charles Darwin University, PO Box 41096, Casuarina, NT 0811, Australia Katja Fischer, Infectious Diseases Program, Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Brisbane, QLD 4029, Australia Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Myogenesis is a complex process in which committed myogenic cells differentiate and fuse into myotubes that mature into the muscle fibres of adult organisms. This process is initiated by a cascade of myogenic regulatory factors expressed upon entry of the cells into the myogenic differentiation programme. However, external signals such as those provided by the extracellular matrix (ECM) are also important in regulating muscle differentiation and morphogenesis. In the present work, we have addressed the role of various ECM substrata on C2C12 myoblast behaviour in vitro. Cells grown on fibronectin align and fuse earlier than cells on laminin or gelatine. Live imaging of C2C12 myoblasts on fibronectin versus gelatine has revealed that fibronectin promotes a directional collective migratory behaviour favouring cell-cell alignment and fusion. We further demonstrate that this effect of fibronectin is mediated by RGD-binding integrins expressed on myoblasts, that N-cadherin contributes to this behaviour, and that it does not involve enhanced myogenic differentiation. Therefore, we suggest that the collective migration and alignment of cells seen on fibronectin leads to a more predictable movement and a positioning that facilitates subsequent fusion of myoblasts. This study highlights the importance of addressing the role of fibronectin, an abundant component of the interstitial ECM during embryogenesis and tissue repair, in the context of myogenesis and muscle regeneration. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1364-1 Authors Raquel Vaz, Centro de Biologia Ambiental/Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal Gabriel G. Martins, Centro de Biologia Ambiental/Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal Sólveig Thorsteinsdóttir, Centro de Biologia Ambiental/Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal Gabriela Rodrigues, Centro de Biologia Ambiental/Departamento de Biologia Animal, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Serine proteases, cysteine proteases, aspartic proteases and matrix metalloproteinases play an essential role in extracellular matrix remodeling and turnover through their proteolytic action on collagens, proteoglycans, fibronectin, elastin and laminin. Proteases can also act on chemokines, receptors and anti-microbial peptides, often potentiating their activity. The intestinal mucosa is the largest interface between the external environment and the tissues of the human body and is constantly exposed to proteolytic enzymes from many sources, including bacteria in the intestinal lumen, fibroblasts and immune cells in the lamina propria and enterocytes. Controlled proteolytic activity is crucial for the maintenance of gut immune homeostasis, for normal tissue turnover and for the integrity of the gut barrier. However, in intestinal immune-mediated disorders, pro-inflammatory cytokines induce the up-regulation of proteases, which become the end-stage effectors of mucosal damage by destroying the epithelium and basement membrane integrity and degrading the extracellular matrix of the lamina propria to produce ulcers. Protease-mediated barrier disruption in turn results in increased amounts of antigen crossing into the lamina propria, driving further immune responses and sustaining the inflammatory process. Content Type Journal Article Category Review Pages 1-12 DOI 10.1007/s00441-012-1390-z Authors Paolo Biancheri, First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, University of Pavia, Piazzale Golgi 5, 27100 Pavia, Italy Antonio Di Sabatino, First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, University of Pavia, Piazzale Golgi 5, 27100 Pavia, Italy Gino R. Corazza, First Department of Medicine, Fondazione IRCCS Policlinico S. Matteo, University of Pavia, Piazzale Golgi 5, 27100 Pavia, Italy Thomas T. MacDonald, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, London, UK Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Fertilization is the process by which male and female haploid gametes (sperm and egg) unite to produce a genetically distinct individual. In mammals, fertilization involves a number of sequential steps, including sperm migration through the female genital tract, sperm penetration through the cumulus mass, sperm adhesion and binding to the zona pellucida, acrosome exocytosis, sperm penetration through the zona and fusion of the sperm and egg plasma membranes. However, freshly ejaculated sperm are not capable of fertilizing an oocyte. They must first undergo a series of biochemical and physiological changes, collectively known as capacitation, before acquiring fertilizing capabilities. Several molecules are required for successful capacitation and in vitro fertilization; these include bicarbonate, serum albumin (normally bovine serum albumin, BSA) and Ca 2+ . Bicarbonate activates the sperm protein soluble adenylyl cyclase (SACY), which results in increased levels of cAMP and cAMP-dependent protein kinase (PKA) activation. The response to bicarbonate is fast and cAMP levels increase within 60 s followed by an increase in PKA activity. Several studies with an anti-phospho-PKA substrate antibody have demonstrated a rapid increase in protein phosphorylation in human, mouse and boar sperm. The target proteins of PKA are not known and the precise role of BSA during capacitation is unclear. Most of the studies provide support for the idea that BSA acts by removing cholesterol from the sperm. The loss of cholesterol has been suggested to affect the bilayer of the sperm plasma membrane making it more fusogenic. The relationship between cholesterol loss and the activation of the cAMP/PKA pathway is also unclear. During early stages of capacitation, Ca 2+ might be involved in the stimulation of SACY, although definitive proof is lacking. Protein tyrosine phosphorylation is another landmark of capacitation but occurs during the late stages of capacitation on a different time-scale from cAMP/PKA activation. Additionally, the tyrosine kinases present in sperm are not well characterized. Although protein phosphorylation depends upon the balanced action of protein kinases and protein phosphatase, we have even less information regarding the role of protein phosphatases during sperm capacitation. Over the last few years, several reports have pointed out that the ubiquitin-proteasome system might play a role during sperm capacitation, acrosome reaction and/or sperm-egg fusion. In the present review, we summarize the information regarding the role of protein kinases, phosphatases and the proteasome during sperm capacitation. Where appropriate, we give examples of the way that these molecules interact and regulate each other’s activities. Content Type Journal Article Category Review Pages 1-18 DOI 10.1007/s00441-012-1370-3 Authors Janetti Signorelli, Biomedical Department, Faculty of Health Sciences, University of Antofagasta, Antofagasta, Chile, Avda. Angamos 601, Antofagasta, Chile Emilce S. Diaz, Biomedical Department, Faculty of Health Sciences, University of Antofagasta, Antofagasta, Chile, Avda. Angamos 601, Antofagasta, Chile Patricio Morales, Biomedical Department, Faculty of Health Sciences, University of Antofagasta, Antofagasta, Chile, Avda. Angamos 601, Antofagasta, Chile Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Mechanisms underlying the unique survival property of human spiral neurons are yet to be explored. P75 (p75 NTR ) is a low affinity receptor for neurotrophins and is known to interact with Trk receptors to modulate ligand binding and signaling. Up-regulation of this receptor was found to be associated with apoptosis as well as with cell proliferation. Its distribution and injury-induced change in expression pattern in the cochlea have been mainly studied in rodents. There is still no report concerning p75 NTR in post-natal human inner ear. We analyzed, for the first time, p75 NTR expression in five freshly fixed human cochleae by using immunohistochemistry techniques, including myelin basic protein (MBP) as a myelin sheath marker and TrkB as the human spiral neuron marker, and by using thin optical sectioning of laser confocal microscopy. The inner ear specimens were obtained from adult patients who had normal pure tone thresholds before the surgical procedures, via a trans-cochlear approach for removal of giant posterior cranial fossa meningioma. The expression of p75 NTR was investigated and localized in the glial cells, including Schwann cells and satellite glial cells in the Rosenthal canal, in the central nerve bundles within the modiolus, and in the osseous spiral lamina of the human cochleae. The biological significance of p75 NTR in human cochlea is discussed. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1395-7 Authors Wei Liu, Department of Surgical Sciences, Section of Otolaryngology, Uppsala University Hospital, 751 85 Uppsala, Sweden Rudolf Glueckert, Department of Otolaryngology and University Clinics Innsbruck (TILAK), University of Innsbruck, Anichstrasse 35, 6020 Innsbruck, Austria Anders Kinnefors, Department of Otolaryngology, Uppsala University Hospital, 75185 Uppsala, Sweden Annelies Schrott-Fischer, Department of Otolaryngology, University of Innsbruck, Anichstrasse 35, 6020 Innsbruck, Austria Mario Bitsche, Division of Clinical and Functional Anatomy, Department of Anatomy, Histology and Embryology, University of Innsbruck, Muellerstr. 59, A 6020 Innsbruck, Austria Helge Rask-Andersen, Department of Otolaryngology, Uppsala University Hospital, 75185 Uppsala, Sweden Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Degeneration of the axon is an important step in the pathomechanism of traumatic, inflammatory and degenerative neurological diseases. Increasing evidence suggests that axonal degeneration occurs early in the course of these diseases and therefore represents a promising target for future therapeutic strategies. We review the evidence for axonal destruction from pathological findings and animal models with particular emphasis on neurodegenerative and neurotraumatic disorders. We discuss the basic morphological and temporal modalities of axonal degeneration (acute, chronic and focal axonal degeneration and Wallerian degeneration). Based on the mechanistic concepts, we then delineate in detail the major molecular mechanisms that underlie the degenerative cascade, such as calcium influx, axonal transport, protein aggregation and autophagy. We finally concentrate on putative therapeutic targets based on the mechanistic prerequisites. Content Type Journal Article Category Review Pages 1-23 DOI 10.1007/s00441-012-1362-3 Authors Paul Lingor, Department of Neurology, University Medicine Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany Jan C. Koch, Department of Neurology, University Medicine Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany Lars Tönges, Department of Neurology, University Medicine Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany Mathias Bähr, Department of Neurology, University Medicine Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Spinal cord injury (SCI) causes the irreversible loss of spinal cord parenchyma including astroglia, oligodendroglia and neurons. In particular, severe injuries can lead to an almost complete neural cell loss at the lesion site and structural and functional recovery might only be accomplished by appropriate cell and tissue replacement. Stem cells have the capacity to differentiate into all relevant neural cell types necessary to replace degenerated spinal cord tissue and can now be obtained from virtually any stage of development. Within the last two decades, many in vivo studies in small animal models of SCI have demonstrated that stem cell transplantation can promote morphological and, in some cases, functional recovery via various mechanisms including remyelination, axon growth and regeneration, or neuronal replacement. However, only two well-documented neural-stem-cell-based transplantation strategies have moved to phase I clinical trials to date. This review aims to provide an overview about the current status of preclinical and clinical neural stem cell transplantation and discusses future perspectives in the field. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-012-1363-2 Authors Beatrice Sandner, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200a, 69121 Heidelberg, Germany Peter Prang, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200a, 69121 Heidelberg, Germany Francisco J. Rivera, Institute of Molecular Regenerative Medicine, Paracelsus Medical University, Salzburg, Austria Ludwig Aigner, Institute of Molecular Regenerative Medicine, Paracelsus Medical University, Salzburg, Austria Armin Blesch, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200a, 69121 Heidelberg, Germany Norbert Weidner, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200a, 69121 Heidelberg, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Implantation and the establishment of pregnancy in mammals involves an intricate interplay of hormones, cytokines, growth factors, proteins, lipids, ions and the extracellular matrix between the uterine epithelium, stroma, immune cells and the conceptus trophectoderm. The divergent nature of implantation in the mouse, human and pig provides not only an interesting contrast in the establishment of pregnancy and early embryonic development but also intriguing similarities with regard to early endometrial-conceptus signaling. An interesting pro-inflammatory cytokine expressed in a number of mammalian species during the period of implantation is interleukin-1β (IL1B). The presence of IL1B might be involved with immunotolerance at the maternal-placental interface and has been proposed as one of the mediators in placental viviparity. The production of IL1B and other proinflammatory cytokines might play a role in establishing pregnancy through modulation of the nuclear factor kappa-B (NFKB) system in a number of species. A model for the regulation of cellular progesterone receptor expression and NFKB activation for endometrial receptivity and conceptus attachment is continuing to evolve and is discussed in the present review. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-012-1356-1 Authors Rodney Geisert, Division of Animal Sciences, University of Missouri, Columbia, Mo., USA Asgerally Fazleabas, Department of Obstetrics and Gynecology, and Reproductive Biology, Michigan State University, East Lansing, Mich., USA Mathew Lucy, Division of Animal Sciences, University of Missouri, Columbia, Mo., USA Daniel Mathew, Division of Animal Sciences, University of Missouri, Columbia, Mo., USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Sudden death in athletes occurs because of the existence of hidden cardiovascular disorders which, during effort, may jeopardize the electrical stability of the heart, triggering ventricular tachycardia and/or fibrillation. Apart from rare conditions of ion channel diseases in the setting of a structurally normal heart, in which the disorder may be easily diagnosed on basal or stress test ECG, cardiac abnormalities at risk of causing sudden death may affect the aorta (Marfan syndrome), the coronary arteries (congenital coronary artery anomalies, premature coronary atherosclerosis), the myocardium (hypertrophic and arrhythmogenic cardiomyopathy), the valves (bicuspid aortic valve, mitral valve prolapse) and the conduction system (pre-excitation syndromes). These structural heart disorders may be detected by ECG and/or echo. The employment of these tools at pre-participation screening can help to identify concealed anomalies, which may play a major role in early diagnosis, risk stratification, and prevention of sudden death. Content Type Journal Article Category Review Pages 1-4 DOI 10.1007/s00441-012-1398-4 Authors Gaetano Thiene, University of Padua Medical School, Padua, Italy Domenico Corrado, University of Padua Medical School, Padua, Italy Ilaria Rigato, University of Padua Medical School, Padua, Italy Cristina Basso, University of Padua Medical School, Padua, Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    In the past decade, an avalanche of findings and reports has correlated arrhythmogenic ventricular cardiomyopathies (ARVC) and Naxos and Carvajal diseases with certain mutations in protein constituents of the special junctions connecting the polar regions (intercalated disks) of mature mammalian cardiomyocytes. These molecules, apparently together with some specific cytoskeletal proteins, are components of (or interact with) composite junctions. Composite junctions contain the amalgamated fusion products of the molecules that, in other cell types and tissues, occur in distinct separate junctions, i.e. desmosomes and adherens junctions. As the pertinent literature is still in an expanding phase and is obviously becoming important for various groups of researchers in basic cell and molecular biology, developmental biology, histology, physiology, cardiology, pathology and genetics, the relevant references so far recognized have been collected and are presented here in the following order: desmocollin-2 (Dsc2, DSC2), desmoglein-2 (Dsg2, DSG2), desmoplakin (DP, DSP), plakoglobin (PG, JUP), plakophilin-2 (Pkp2, PKP2) and some non-desmosomal proteins such as transmembrane protein 43 (TMEM43), ryanodine receptor 2 (RYR2), desmin, lamins A and C, striatin, titin and transforming growth factor-β3 (TGFβ3), followed by a collection of animal models and of reviews, commentaries, collections and comparative studies. Content Type Journal Article Category Mini Review Pages 1-9 DOI 10.1007/s00441-012-1365-0 Authors Steffen Rickelt, Helmholtz Group for Cell Biology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 581, Building TP4, 69120 Heidelberg, Germany Sebastian Pieperhoff, BHF Centre for Cardiovascular Science, The Queen’s Medical Research Institute, University of Edinburgh, 47 Little France Crescent, EH164TJ Edinburgh, Scotland, UK Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The delivery of newly-formed secretory content to the granule inventory occurs through direct fusion of recently formed granules and mature granules. The introduction of knockout mice allowed us to investigate the characteristics of the delivery process and to determine the core protein machinery required for granule growth. The SNARE machinery mediates membrane fusion and is essential for the granule lifecycle. In the current work, we use VAMP8 knockout mice to show that the SNARE machinery plays a critical role in the process of granule homotypic fusion. Consistent with this, the mutated mouse pancreatic acinar secretory granules are significantly smaller compared to the control group, demonstrating few granule profiles that might be the result of homotypic fusion. Content Type Journal Article Category Regular Article Pages 1-6 DOI 10.1007/s00441-012-1400-1 Authors Ilan Hammel, Department of Pathology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978 Israel Cheng-Chun Wang, Membrane Biology Laboratory, Institute of Molecular and Cell Biology, 61 Biopolis Dr., Singapore, 138673 Singapore Wanjin Hong, Membrane Biology Laboratory, Institute of Molecular and Cell Biology, 61 Biopolis Dr., Singapore, 138673 Singapore Dina Amihai, Department of Pathology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978 Israel Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The kidney filtration barrier consists of the capillary endothelium, the glomerular basement membrane and the slit diaphragm localized between foot processes of neighbouring podocytes. We report that collagen XVII, a transmembrane molecule known to be required for epithelial adhesion, is expressed in podocytes of normal human and mouse kidneys and in endothelial cells of the glomerular filtration barrier. Immunoelectron microscopy has revealed that collagen XVII is localized in foot processes of podocytes and in the glomerular basement membrane. Its role in kidney has been analysed in knockout mice, which survive to birth but have high neonatal mortality and skin blistering and structural abnormalities in their glomeruli. Morphometric analysis has shown increases in glomerular volume fraction and surface densities of knockout kidneys, indicating an increased glomerular amount in the cortex. Collagen XVII deficiency causes effacement of podocyte foot processes; however, major slit diaphragm disruptions have not been detected. The glomerular basement membrane is split in areas in which glomerular and endothelial basement membranes meet. Differences in the expression of collagen IV, integrins α3 or β1, laminin α5 and nephrin have not been observed in mutant mice compared with controls. We propose that collagen XVII has a function in the attachment of podocyte foot processes to the glomerular basement membrane. It probably contributes to podocyte maturation and might have a role in glomerular filtration. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1368-x Authors Tiina Hurskainen, Department of Dermatology, Oulu Center for Cell-Matrix Research, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Jyri Moilanen, Department of Dermatology, Oulu Center for Cell-Matrix Research, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Raija Sormunen, Biocenter Oulu, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Claus-Werner Franzke, Department of Dermatology, University Medical Center Freiburg and Freiburg Institute of Advanced Studies, School of Life Sciences, 79104 Freiburg, Germany Raija Soininen, Biocenter Oulu, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Stefanie Loeffek, Department of Dermatology, University Medical Center Freiburg and Freiburg Institute of Advanced Studies, School of Life Sciences, 79104 Freiburg, Germany Laura Huilaja, Department of Dermatology, Oulu Center for Cell-Matrix Research, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Matti Nuutinen, Department of Paediatrics, Oulu University Hospital, Kajaanintie 52, FIN-90220 Oulu, Finland Leena Bruckner-Tuderman, Department of Dermatology, University Medical Center Freiburg and Freiburg Institute of Advanced Studies, School of Life Sciences, 79104 Freiburg, Germany Helena Autio-Harmainen, Department of Pathology, University of Oulu, Oulu University Hospital, Aapistie 3, FIN-90220 Oulu, Finland Kaisa Tasanen, Department of Dermatology, Oulu Center for Cell-Matrix Research, University of Oulu, Aapistie 5A, FIN-90220 Oulu, Finland Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Estrogens and androgens play important roles in regulating the hormone-secreting functions of the pituitary gland by binding to their corresponding receptors. However, the expression of estrogen receptors (ERs) and the androgen receptor (AR) and the cell types containing ERs and AR in the anterior pituitary gland of adult chickens have not been well-studied. In this study, the distribution of ERα, AR and their corresponding cell types in the anterior pituitary gland of adult cockerels was detected by immunohistochemistry. The results showed that ERα was expressed in 68.63 % of luteinizing hormone (LH) producing cells but was not found in thyrotropes, lactotropes, somatotropes, corticotropes and folliculo-stellate (FS) cells. Pituitary hormone and AR double labeling results showed that about 37 % of LH cells and 50 % of thyroid-stimulating hormone (TSH) producing cells expressed AR, respectively. In contrast, less than 1 % of the somatotropes had an AR positive signal and AR signals were not detected in lactotropes, corticotropes or FS cells. In addition, there were only a few AR and ERα dual-labeled cells observed. These novel results provide evidence for a cell-specific distribution of ERα and AR in the anterior pituitary from adult cockerels by immunohistochemistry. The different distributions of ERα and AR in the LH cells suggest that the feedback-regulating mechanisms of estrogen and androgen on the pituitary hormones secretion are different. The functions and related mechanisms still need to be elucidated further. Content Type Journal Article Category Regular Article Pages 1-8 DOI 10.1007/s00441-012-1399-3 Authors Dehao Sun, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193 People’s Republic of China Tongtong Cui, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193 People’s Republic of China Haoshu Luo, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193 People’s Republic of China Ruiguo Li, China National Center for Biotechnology Development, Beijing, 100036 People’s Republic of China Sheng Cui, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193 People’s Republic of China Jiali Liu, State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193 People’s Republic of China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Mesenchymal stem cells (MSCs) are multipotent cells with the capacity for self-renewal and differentiation and have a broad tissue distribution. These characteristics make them candidate cells for wound healing and regeneration in a variety of disorders. Endogenous MSCs or exogenously delivered MSCs can traffic and migrate to injured tissue and participate in the healing of this tissue. The concentrated conditioned medium from MSCs can modulate wound repair without MSCs being present in the wound. The therapeutic effects of MSCs might be attributable to their ability to differentiate and transdifferentiate into tissue-specific cells, to fuse with the resident cells, to secrete a wide array of paracrine factors in order to stimulate the survival and functional recovery of the resident cells, or to regulate the local microenviroment or niche and immune response. These mechanisms are probably independent but not mutually exclusive. In many circumstances, a combination of these protective mechanisms might work together to affect cutaneous wound healing. This review gives a brief overview and discusses the mechanisms by which MSCs promote skin repair and regeneration, although the specific mechanisms in each type of cutaneous wound are still unclear and controversial. A comprehensive understanding of the mechanisms should allow us to find advanced and better treatment strategies for various skin diseases, even those that are currently incurable. Content Type Journal Article Category Review Pages 1-7 DOI 10.1007/s00441-012-1393-9 Authors HaiHong Li, Department of Burn and Plastic Surgery, The Second Affiliated Hospital, Shantou University Medical College, ShanTou, GuangDong Province 515041, People’s Republic of China Xiaobing Fu, Burns Institute, The First Affiliated Hospital, Chinese PLA General Hospital, Trauma Center of Postgraduate Medical School, 51 Fu Cheng Road, Beijing, 100048 People’s Republic of China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Perineuronal nets (PNNs) are reticular structures that surround the cell body of many neurones, and extend along their dendrites. They are considered to be a specialized extracellular matrix in the central nervous system (CNS). PNN formation is first detected relatively late in development, as the mature synaptic circuitry of the CNS is established and stabilized. Its unique distribution in different CNS regions, the timing of its establishment, and the changes it undergoes after injury all point toward diverse and important functions that it may be performing. The involvement of PNNs in neuronal plasticity has been extensively studied over recent years, with developmental, behavioural, and functional correlations. In this review, we will first briefly detail the structure and organization of PNNs, before focusing our discussion on their unique roles in neuronal development and plasticity. The PNN is an important regulator of CNS plasticity, both during development and into adulthood. Production of critical PNN components is often triggered by appropriate sensory experiences during early postnatal development. PNN deposition around neurones helps to stabilize the established neuronal connections, and to restrict the plastic changes due to future experiences within the CNS. Disruption of PNNs can reactivate plasticity in many CNSs, allowing activity-dependent changes to once again modify neuronal connections. The mechanisms through which PNNs restrict CNS plasticity remain unclear, although recent advances promise to shed additional light on this important subject. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-012-1375-y Authors Difei Wang, Centre for Brain Repair, University of Cambridge, Robinson Way, Cambridge, CB2 0PY UK James Fawcett, Centre for Brain Repair, University of Cambridge, Robinson Way, Cambridge, CB2 0PY UK Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Axonal regeneration is inhibited by a plethora of different mechanisms in the adult central nervous system (CNS). While neurotrophic factors have been shown to stimulate axonal growth in numerous animal models of nervous system injury, a lack of suitable growth substrates, an insufficient activation of neuron-intrinsic regenerative programs, and extracellular inhibitors of regeneration limit the efficacy of neurotrophic factor delivery for anatomical and functional recovery after spinal cord injury. Thus, growth-stimulating factors will likely have to be combined with other treatment approaches to tap into the full potential of growth factor therapy for axonal regeneration. In addition, the temporal and spatial distribution of growth factors have to be tightly controlled to achieve biologically active concentrations, to allow for the chemotropic guidance of axons, and to prevent adverse effects related to the widespread distribution of neurotrophic factors. Here, we will review the rationale for combinatorial treatments in axonal regeneration and summarize some recent progress in promoting axonal regeneration in the injured CNS using such approaches. Content Type Journal Article Category Review Pages 1-11 DOI 10.1007/s00441-012-1388-6 Authors Julianne McCall, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200 a, 69118 Heidelberg, Germany Norbert Weidner, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200 a, 69118 Heidelberg, Germany Armin Blesch, Spinal Cord Injury Center, Heidelberg University Hospital, Schlierbacher Landstrasse 200 a, 69118 Heidelberg, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Adipose-derived stromal cells (ASC) are promising candidates for cell therapy, for example to treat myocardial infarction. Commonly, fetal bovine serum (FBS) is used in ASC culturing. However, FBS has several disadvantages. Its effects differ between batches and, when applied clinically, transmission of pathogens and antibody development against FBS are possible. In this study, we investigated whether FBS can be substituted by human platelet lysate (PL) in ASC culture, without affecting functional capacities particularly important for cardiac repair application of ASC. We found that PL-cultured ASC had a significant 3-fold increased proliferation rate and a significantly higher attachment to tissue culture plastic as well as to endothelial cells compared with FBS-cultured ASC. PL-cultured ASC remained a significant 25% smaller than FBS-cultured ASC. Both showed a comparable surface marker profile, with the exception of significantly higher levels of CD73, CD90, and CD166 on PL-cultured ASC. PL-cultured ASC showed a significantly higher migration rate compared with FBS-cultured ASC in a transwell assay. Finally, FBS- and PL-cultured ASC had a similar high capacity to differentiate towards cardiomyocytes. In conclusion, this study showed that culturing ASC is more favorable in PL-supplemented medium compared with FBS-supplemented medium. Content Type Journal Article Category Regular Article Pages 119-130 DOI 10.1007/s00441-012-1360-5 Authors B. A. Naaijkens, Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands H. W. M. Niessen, Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands H-J. Prins, Department of Oral & Maxillofacial Surgery, VU University Medical Center, Amsterdam, The Netherlands P. A. J. Krijnen, Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands T. J. A. Kokhuis, Interuniversity Cardiology Institute of the Netherlands (ICIN), Utrecht, The Netherlands N. de Jong, Interuniversity Cardiology Institute of the Netherlands (ICIN), Utrecht, The Netherlands V. W. M. van Hinsbergh, Department of Physiology, VU University Medical Center, Amsterdam, The Netherlands O. Kamp, Department of Cardiology, VU University Medical Center, Amsterdam, The Netherlands M. N. Helder, Department of Orthopaedics, VU University Medical Center, Amsterdam, The Netherlands R. J. P. Musters, Department of Physiology, VU University Medical Center, Amsterdam, The Netherlands A. van Dijk, Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands L. J. M. Juffermans, Department of Physiology, VU University Medical Center, Amsterdam, The Netherlands Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Both the uterus and vagina develop from the Müllerian duct but are quite distinct in morphology and function. To investigate factors controlling epithelial differentiation and cell proliferation in neonatal uterus and vagina, we focused on Hedgehog (HH) signaling. In neonatal mice, Sonic hh ( Shh ) was localized in the vaginal epithelium and Indian hh ( Ihh ) was slightly expressed in the uterus and vagina, whereas all Glioma-associated oncogene homolog ( Gli ) genes were mainly expressed in the stroma. The expression of target genes of HH signaling was high in the neonatal vagina and in the uterus, it increased with growth. Thus, in neonatal mice, Shh in the vaginal epithelium and Ihh in the uterus and vagina activated HH signaling in the stroma. Tissue recombinants showed that vaginal Shh expression was inhibited by the vaginal stroma and uterine Ihh expression was stimulated by the uterine stroma. Addition of a HH signaling inhibitor decreased epithelial cell proliferation in organ-cultured uterus and vagina and increased stromal cell proliferation in organ-cultured uterus. However, it did not affect epithelial differentiation or the expression of growth factors in organ-cultured uterus and vagina. Thus, activated HH signaling stimulates epithelial cell proliferation in neonatal uterus and vagina but inhibits stromal cell proliferation in neonatal uterus. Content Type Journal Article Category Regular Article Pages 239-247 DOI 10.1007/s00441-012-1350-7 Authors Tadaaki Nakajima, Graduate School of Nanobioscience, Yokohama City University, Yokohama, 236-0027 Japan Taisen Iguchi, The Graduate University for Advanced Studies and Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki, 444-8787 Japan Tomomi Sato, Graduate School of Nanobioscience, Yokohama City University, Yokohama, 236-0027 Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Mutation of the human TRPS1 gene leads to trichorhinophalangeal syndrome (TRPS), which is characterized by an abnormal development of various organs including the craniofacial skeleton. Trps1 has recently been shown to be expressed in the jaw joints of zebrafish; however, whether Trps1 is expressed in the mammalian temporomandibular joint (TMJ), or whether it is necessary for TMJ development is unknown. We have analyzed (1) the expression pattern of Trps1 during TMJ development in mice and (2) TMJ development in Trps1 knockout animals. Trps1 is expressed in the maxillo-mandibular junction at embryonic day (E) 11.5. At E15.5, expression is restricted to the developing condylar cartilage and to the surrounding joint disc progenitor cells. In Trps1 knockout mice, the glenoid fossa of the temporal bone forms relatively normally but the condylar process is extremely small and the joint disc and cavities do not develop. The initiation of condyle formation is slightly delayed in the mutants at E14.5; however, at E18.5, the flattened chondrocyte layer is narrowed and most of the condylar chondrocytes exhibit precocious chondrocyte maturation. Expression of Runx2 and its target genes is expanded toward the condylar apex in the mutants. These observations underscore the indispensable role played by Trps1 in normal TMJ development in supporting the differentiation of disc and synoviocyte progenitor cells and in coordinating condylar chondrocyte differentiation. Content Type Journal Article Category Regular Article Pages 131-140 DOI 10.1007/s00441-012-1372-1 Authors Ikumi Michikami, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Toshiya Fukushi, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Shiho Honma, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Seisuke Yoshioka, Department of Restorative Dentistry, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Shunji Itoh, First Department of Pathology, Wakayama Medical University Medical School, 811-1 Kimiidera, Wakayama, 641-0012 Japan Yasuteru Muragaki, First Department of Pathology, Wakayama Medical University Medical School, 811-1 Kimiidera, Wakayama, 641-0012 Japan Kojiro Kurisu, Department of Judo Therapy, Yukioka School of Allied Health Professions, 2-2-11 Kitaku, Osaka, 530-0021 Japan Takashi Ooshima, Department of Pediatric Dentistry, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Satoshi Wakisaka, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Makoto Abe, Department of Oral Anatomy and Developmental Biology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871 Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a clinically and genetically heterogeneous heart muscle disorder associated with ventricular arrhythmias and risk of sudden death. The disease is heredo-familial, and mutations in desmosomal genes have been identified in about half of patients. Recent experimental models confirm this disease develops after birth due to progressive myocardial dystrophy. Genotype–phenotype correlations, including magnetic resonance and pathology studies on heart specimens, are currently demonstrating that the spectrum of the disease is wider than initially thought and usually referred to with the adjective “right ventricular”, with the evidence of biventricular or even isolated left ventricular forms, so that it is increasingly identified simply as “arrhythmogenic cardiomyopathy”. A revision of the diagnostic criteria encompassing familial, electrocardiographic, arrhythmic, morpho-functional and histopathologic findings, has been made to improve diagnostic sensitivity and specificity, in particular of the concealed forms and left-dominant subtypes of the disease. Experimental models are mandatory to gain an insight into the cascade of cellular and molecular events leading from gene defect to myocardial dystrophy in ARVC. Content Type Journal Article Category Mini Review Pages 1-5 DOI 10.1007/s00441-012-1402-z Authors Stefania Rizzo, Department of Cardiac, Thoracic and Vascular Sciences, University of Padua Medical School, Via Gabelli 61, 35121 Padua, Italy Kalliopi Pilichou, Department of Cardiac, Thoracic and Vascular Sciences, University of Padua Medical School, Via Gabelli 61, 35121 Padua, Italy Gaetano Thiene, Department of Cardiac, Thoracic and Vascular Sciences, University of Padua Medical School, Via Gabelli 61, 35121 Padua, Italy Cristina Basso, Department of Cardiac, Thoracic and Vascular Sciences, University of Padua Medical School, Via Gabelli 61, 35121 Padua, Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The ability of injured peripheral nerves to regenerate and reinnervate their original targets is a characteristic feature of the peripheral nervous system (PNS). On the other hand, neurons of the central nervous system (CNS), including retinal ganglion cell (RGC) axons, are incapable of spontaneous regeneration. In the adult PNS, axonal regeneration after injury depends on well-orchestrated cellular and molecular processes that comprise a highly reproducible series of degenerative reactions distal to the site of injury. During this fine-tuned process, named Wallerian degeneration , a remodeling of the distal nerve fragment prepares a permissive microenvironment that permits successful axonal regrowth originating from the proximal nerve fragment. Therefore, a multitude of adjusted intrinsic and extrinsic factors are important for surviving neurons, Schwann cells, macrophages and fibroblasts as well as endothelial cells in order to achieve successful regeneration. The aim of this review is to summarize relevant extrinsic cellular and molecular determinants of successful axonal regeneration in rodents that contribute to the regenerative microenvironment of the PNS. Content Type Journal Article Category Review Pages 1-10 DOI 10.1007/s00441-012-1389-5 Authors Frank Bosse, Molecular Neurobiology Laboratory, Department of Neurology, Medical Faculty, Heinrich-Heine-University Düsseldorf, Moorenstr 5, 40225 Düsseldorf, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Spinal and bulbar muscular atrophy (SBMA), also known as Kennedy’s disease, is an adult-onset, X-linked motor neuron disease characterized by muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. SBMA is caused by the expansion of a CAG triplet repeat, encoding a polyglutamine tract within the first exon of the androgen receptor (AR) gene. The histopathological finding in SBMA is the loss of lower motor neurons in the anterior horn of the spinal cord as well as in the brainstem motor nuclei. There is no established disease-modifying therapy for SBMA. Animal studies have revealed that the pathogenesis of SBMA depends on the level of serum testosterone, and that androgen deprivation mitigates neurodegeneration through inhibition of nuclear accumulation and/or stabilization of the pathogenic AR. Heat shock proteins, the ubiquitin–proteasome system and transcriptional regulation are also potential targets for development of therapy for SBMA. Among these therapeutic approaches, the luteinizing hormone-releasing hormone analogue, leuprorelin, prevents nuclear translocation of aberrant AR proteins, resulting in a significant improvement of disease phenotype in a mouse model of SBMA. In a phase 2 clinical trial of leuprorelin, the patients treated with this drug exhibited decreased mutant AR accumulation in scrotal skin biopsy. Phase 3 clinical trial showed the possibility that leuprorelin treatment is associated with improved swallowing function particularly in patients with a disease duration less than 10 years. These observations suggest that pharmacological inhibition of the toxic accumulation of mutant AR is a potential therapy for SBMA. Content Type Journal Article Category Review Pages 1-8 DOI 10.1007/s00441-012-1377-9 Authors Haruhiko Banno, Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Masahisa Katsuno, Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Keisuke Suzuki, Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Fumiaki Tanaka, Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Gen Sobue, Department of Neurology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550 Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The European shore crab Carcinus maenas and the common hermit crab Pagurus bernhardus are members of the sister taxa Brachyura and Anomura (together forming the taxon Meiura) respectively. Both species share similar coastal marine habitats and thus are confronted with similar environmental conditions. This study sets out to explore variations of general brain architecture of species that live in seemingly similar habitats but belong to different major malacostracan taxa and to understand possible differences of sensory systems and related brain compartments. We examined the brains of Carcinus maenas , Pagurus bernhardus , and three other hermit crab species with immunohistochemistry against tyrosinated tubulin, f-actin, synaptic proteins, RF-amides and allatostatin. Our comparison showed that their optic neuropils within the eyestalks display strong resemblance in gross morphology as well as in detailed organization, suggesting a rather similar potential of processing visual input. Besides the well-developed visual system, the olfactory neuropils are distinct components in the brain of both C. maenas and P. bernhardus as well as the other hermit crabs, suggesting that close integration of olfactory and visual information may be useful in turbid marine environments with low visibility, as is typical for many habitats such as, e.g., the Baltic and the North Sea. Comparing the shape of the olfactory glomeruli in the anomurans showed some variations, ranging from a wedge shape to an elongate morphology. Furthermore, the tritocerebrum and the organization of the second antennae associated with the tritocerebrum seem to differ markedly in C. maenas and P. bernhardus , indicating better mechanosensory abilities in the latter close to those of other Decapoda with long second antennae, such as Astacida, Homarida, or Achelata. This aspect may also represent an adaptation to the “hermit lifestyle” in which competition for shells is a major aspect of their life history. The shore crab C. maenas , on the other hand seems to rely much less on mechanosensory information mediated by the second antennae but in water, the visual and the olfactory senses seem to be the most important modalities. Content Type Journal Article Category Regular Article Pages 47-69 DOI 10.1007/s00441-012-1353-4 Authors Jakob Krieger, Zoological Institute and Museum, Department of Cytology and Evolutionary Biology, University of Greifswald, Soldmannstrasse 23, 17487 Greifswald, Germany Andy Sombke, Zoological Institute and Museum, Department of Cytology and Evolutionary Biology, University of Greifswald, Soldmannstrasse 23, 17487 Greifswald, Germany Florian Seefluth, Zoological Institute and Museum, Department of Cytology and Evolutionary Biology, University of Greifswald, Soldmannstrasse 23, 17487 Greifswald, Germany Matthes Kenning, Zoological Institute and Museum, Department of Cytology and Evolutionary Biology, University of Greifswald, Soldmannstrasse 23, 17487 Greifswald, Germany Bill S. Hansson, Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology, Beutenberg Campus, Hans-Knöll-Str. 8, 07745 Jena, Germany Steffen Harzsch, Zoological Institute and Museum, Department of Cytology and Evolutionary Biology, University of Greifswald, Soldmannstrasse 23, 17487 Greifswald, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    The clinical significance of lymphangiogenesis in cervical cancer remains controversial. Our aim was to investigate the correlation between lymphangiogenesis, lymphatic vessel invasion (LVI) and tumor metastasis, invasion and prognosis in squamous cell cervical cancer. Paraffin sections of 90 patients with FIGO (Fédération Internationale de Gynécologie et d'Obstétrique) Ib1-IIa squamous cell cervical cancer were stained for immunohistochemistry with a D2-40 monoclonal antibody against the carcinoembryonic antigen M2A. The lymphatic vessel density (LVD) and LVI were measured, and their relationship with the clinicopathological data was analyzed. D2-40-positive lymphatic vessels were found in 75 of the 90 patients (83.3 %). All D2-40-positive vessels were located in peritumoral areas. The mean±SD of the peritumoral LVD was 10.08±4.16. The positive rate of LVI was 32.0 % (24/75). The recurrence rate of patients with LVD 〉10 (62.1 %, 18/29) was significantly higher than that of patients with LVD ≤10 (34.8 %, 16/46, P  = 0.021). The 5-year recurrence-free survival rate of patients with LVD 〉10 (41.0 %) was significantly lower than that of patients with LVD ≤10 (67.0 %, P  = 0.045). Univariate analysis showed that the peritumoral LVD (≤10 vs 〉10) was correlated with LVI (absent vs present, P  = 0.016). The peritumoral LVD and LVI showed no correlation with age, FIGO stage, tumor size, tumor grade, depth of invasion, or pelvic lymph node metastasis (all: P  〉 0.05). Peritumoral lymphangiogenesis was correlated with the recurrence and recurrence-free survival in patients with squamous cell cervical cancer. Examination of peritumoral LVD in these patients might therefore help to estimate the risk of recurrence. Content Type Journal Article Category Regular Article Pages 1-8 DOI 10.1007/s00441-012-1384-x Authors Ying Xiong, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Li-Ping Cao, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Hui-Lan Rao, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Mu-Yan Cai, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Li-Zhi Liang, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Ji-Hong Liu, State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, People’s Republic of China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    We have investigated whether 5% oxygen tension (O 2 ) is beneficial for neocartilage formation when chondrocytes are cultured in transwell-COL inserts. Six million bovine primary chondrocytes were cultured in an insert with DMEM supplemented with 10% fetal bovine serum and antibiotics, with or without glucosamine sulphate (GS) in a 5% or 20% O 2 environment for 2, 4, or 6 weeks. The samples were collected for the histological staining of proteoglycans (PGs) and type II collagen, quantitative reverse transcription with the polymerase chain reaction (RT-PCR) analyses of the mRNA expression of aggrecan and procollagen α 1 (II), procollagen α 2 (I) and hyaluronan synthase 2, quantitation of PGs, and agarose gel electrophoresis. Neocartilage produced at 20% O 2 appeared larger than that at 5% O 2 . Histological staining showed that more PGs and type II collagen and better native cartilage structure were produced at 20% than at 5% O 2 . The thickness of neocartilage increased during the culture period. Quantitative RT-PCR showed that the procollagen α 1 (II) mRNA expression level was significantly higher at 20% than at 5% O 2 . However, no significant difference in gene expression and PG content was found between control and GS-treated cultures at either 20% or 5% O 2 . Thus, in contrast to monolayer cultures, engineered cartilage from scaffold-free cultured chondrocytes at 20% O 2 produced better extracellular matrix (ECM) than that at 5% O 2 . PGs were mainly large. Exogenous GS was not beneficial for the ECM in scaffold-free chondrocyte cultures. Content Type Journal Article Category Regular Article Pages 109-117 DOI 10.1007/s00441-012-1366-z Authors Chengjuan Qu, Department of Biosciences, Biocenter Kuopio, University of Eastern Finland, PO Box 1627, Yliopistoranta 1F, 70211 Kuopio, Finland Heli Lindeberg, Department of Biosciences, University of Eastern Finland, Kuopio, Finland Janne H. Ylärinne, Department of Biosciences, University of Eastern Finland, Kuopio, Finland Mikko J. Lammi, Department of Biosciences, Biocenter Kuopio, University of Eastern Finland, PO Box 1627, Yliopistoranta 1F, 70211 Kuopio, Finland Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    The Na + -K + -2Cl - cotransporter-2 (NKCC2) has long been recognized as a “kidney-specific” transporter and is important in salt reabsorption. NKCC2 has been found in the gastric mucosa; however, its cellular distribution and function remain obscure. The present study characterized the distribution pattern of NKCC2 in mammalian gastric mucosa and investigated its response to osmotic challenge. Reverse transcription with the polymerase chain reaction, Western blot and immunofluorescence were used to determine NKCC2 expression and localization. The effect of osmotic shock on NKCC2 expression was studied in isolated gastric mucosa with variable osmolarity treatment. Results from all of the above studies were compared with those of NKCC1. Our data indicated that NKCC1 and NKCC2 were expressed in the gastric mucosa of rat, mouse and human. The mRNA transcripts and proteins for NKCC1 and NKCC2 were broadly expressed in the rat gastric mucosa. In rat and mouse, NKCC1 was largely confined to the lower part of the oxyntic and pyloric gland areas, whereas NKCC2 extended throughout the gastric glands. NKCC1 immunoreactivity was strongly expressed in the parietal and chief cells but was weaker in the mucous cells. NKCC2 was abundantly located in the parietal and mucous cells but faintly distributed in the chief cells. Hypertonic treatment increased the protein level of NKCC1 and caused evident membrane translocation. In contrast, NKCC2 was significantly downregulated and no obvious membrane translocation was observed. Thus, NKCC2 displayed a more ubiquitous distribution in the gastric mucosa and might work coordinately with NKCC1 to maintain cell volume homeostasis under hypertonic conditions. Content Type Journal Article Category Regular Article Pages 155-165 DOI 10.1007/s00441-012-1359-y Authors T. Ji, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China S. Liu, Department of Biology, College of Science and Health, University of Wisconsin - La Crosse, La Crosse, Wis., USA L. F. Zheng, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China Q. Wang, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China Z. F. Dou, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China Y. Zhang, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China J. X. Zhu, Department of Physiology, School of Basic Medical Sciences, Capital Medical University, No.10 Xitoutiao, You An Men, Beijing, 100069 China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Analysis of transcription factor function during neurogenesis has provided a huge amount of data on the generation and specification of diverse neuron populations in the central and peripheral nervous systems of vertebrates. However, an understanding of the induction of key neuron functions including electrical information processing and synaptic transmission lags seriously behind. Whereas pan-neuronal markers such as neurofilaments, neuron-specific tubulin and RNA-binding proteins have often been included in developmental analysis, the molecular players underlying electrical activity and transmitter release have been neglected in studies addressing gene expression during neuronal induction. Here, I summarize the evidence for a distinct accumulation pattern of mRNAs for synaptic proteins, a pattern that is delayed compared with pan-neuronal gene expression during neurogenesis. The conservation of this pattern across diverse avian and mammalian neuron populations suggests a common mechanism for the regulation of various sets of neuronal genes during initial neuronal differentiation. The co-regulation of genes coding for synaptic proteins from embryonic to postnatal development indicates that the expression of the players required for synaptic transmission shares common regulatory features. For the ion channels involved in neuronal electrical activity, such as voltage-gated sodium channels, the situation is less clear because of the lack of comparative studies early during neurogenesis. Transcription factors have been characterized that regulate the expression of synaptic proteins in vitro and in vivo. They currently do not explain the co-regulation of these genes across different neuron populations. The neuron-restrictive silencing factor NRSF/REST targets a large gene set, but not all of the genes coding for pan-neuronal, synaptic and ion channel proteins. The discrepancy between NRSF/REST loss-of-function and silencer-to-activator-switch studies leaves the full functional implications of this factor open. Together with microRNAs, splicing regulators, chromatin remodellers and an increasing list of transcriptional regulators, the factor is embedded in feedback circuits with the potential to orchestrate neuronal differentiation. The precise regulation of the coordinated expression of proteins underlying key neuronal functions by these circuits during neuronal induction is a major emerging topic. Content Type Journal Article Category Review Pages 1-27 DOI 10.1007/s00441-012-1367-y Authors Uwe Ernsberger, Max Planck Institute for Brain Research, Deutschordenstrasse 46, 60528 Frankfurt, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Human dental pulp contains adult stem cells. Our recent study demonstrated the localization of putative dental pulp stem/progenitor cells in the rat developing molar by chasing 5-bromo-2’-deoxyuridine (BrdU)-labeling. However, there are no available data on the localization of putative dental pulp stem/progenitor cells in the mouse molar. This study focuses on the mapping of putative dental pulp stem/progenitor cells in addition to the relationship between cell proliferation and differentiation in the developing molar using BrdU-labeling. Numerous proliferating cells appeared in the tooth germ and the most active cell proliferation in the mesenchymal cells occurred in the prenatal stages, especially on embryonic Day 15 (E15). Cell proliferation in the pulp tissue dramatically decreased in number by postnatal Day 3 (P3) when nestin-positive odontoblasts were arranged in the cusped areas and disappeared after postnatal Week 1 (P1W). Root dental papilla included numerous proliferating cells during P5 to P2W. Three to four intraperitoneal injections of BrdU were given to pregnant ICR mice and revealed slow-cycling long-term label-retaining cells (LRCs) in the mature tissues of postnatal animals. Numerous dense LRCs postnatally decreased in number and reached a plateau after P1W when they mainly resided in the center of the dental pulp, associating with blood vessels. Furthermore, numerous dense LRCs co-expressed mesenchymal stem cell markers such as STRO-1 and CD146. Thus, dense LRCs in mature pulp tissues were believed to be dental pulp stem/progenitor cells harboring in the perivascular niche surrounding the endothelium. Content Type Journal Article Category Regular Article Pages 95-107 DOI 10.1007/s00441-012-1347-2 Authors Yuko Ishikawa, Division of Oral Science for Health Promotion, Department of Oral Health and Welfare, Niigata University Graduate School of Medical and Dental Sciences, 2-5274 Gakkocho-dori, Chuo-ku, Niigata, 951-8514 Japan Hiroko Ida-Yonemochi, Division of Anatomy and Cell Biology of the Hard Tissue, Department of Tissue Regeneration and Reconstruction, Niigata University Graduate School of Medical and Dental Sciences, 2-5274 Gakkocho-dori, Chuo-ku, Niigata, 951-8514 Japan Kuniko Nakakura-Ohshima, Pediatric Dentistry, Niigata University Medical and Dental Hospital, 2-5274 Gakkocho-dori, Chuo-ku, Niigata, 951-8514 Japan Hayato Ohshima, Division of Anatomy and Cell Biology of the Hard Tissue, Department of Tissue Regeneration and Reconstruction, Niigata University Graduate School of Medical and Dental Sciences, 2-5274 Gakkocho-dori, Chuo-ku, Niigata, 951-8514 Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X Journal Volume Volume 348 Journal Issue Volume 348, Number 1

Description:    Zinc ions in the synaptic vesicles of zinc-enriched neurons (ZEN) seem to have an important role in normal physiological and pathophysiological processes in target organ innervation. The factor directly responsible for the transport of zinc ions into synaptic vesicles is zinc transporter 3 (ZnT3), a member of the divalent cation zinc transporters and an excellent marker of ZEN neurons. As data concerning the existence of ZEN neurons in the small intestine is lacking, this study was designed to disclose the presence and neurochemical coding of such neurons in the porcine jejunum. Cryostat sections (10 mμ thick) of porcine jejunum were processed for routine double- and triple-immunofluorescence labeling for ZnT3 in various combinations with immunolabeling for other neurochemicals including pan-neuronal marker (PGP9.5), substance P (SP), somatostatin (SOM), vasoactive intestinal peptide (VIP), nitric oxide synthase (NOS), leu-enkephalin (LENK), vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), galanin (GAL), and calcitonin-gene related peptide (CGRP). Immunohistochemistry revealed that approximately 39%, 49%, and 45% of all PGP9.5- positive neurons in the jejunal myenteric (MP), outer submucous (OSP), and inner submucous (ISP) plexuses, respectively, were simultaneously ZnT3 + . The majority of ZnT3 + neurons in all plexuses were also VAChT-positive. Both VAChT-positive and VAChT-negative ZnT3 + neurons co-expressed a variety of active substances with diverse patterns of co-localization depending on the plexus studied. In the MP, the largest populations among both VAChT-positive and VAChT-negative ZnT3 + neurons were NOS-positive cells. In the OSP and ISP, substantial subpopulations of ZnT3 + neurons were VAChT-positive cells co-expressing SOM and GAL, respectively. The broad-spectrum of active substances that co-localize with the ZnT3 + neurons in the porcine jejunum suggests that ZnT3 takes part in the regulation of various processes in the gut, both in normal physiological and during pathophysiological processes. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1486-5 Authors Joanna Wojtkiewicz, Faculty Medical Sciences, Department of Neurology and Neurosurgery, Stem Cell Research Laboratory, University of Warmia and Mazury, ul. Warszawska 30, 10-082 Olsztyn, Poland Maciej Równiak, Department of Comparative Anatomy, Faculty of Biology, University of Warmia and Mazury, Olsztyn, Poland Robert Crayton, Department and Clinic of Urology, Faculty of Medical Sciences, Medical University of Warsaw, Warsaw, Poland Mariusz Majewski, Department of Human Physiology, Faculty of Medical Sciences, University of Warmia and Mazury, Olsztyn, Poland Sławomir Gonkowski, Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury, Olsztyn, Poland Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Flatfish metamorphosis is the most dramatic post-natal developmental event in teleosts. Thyroid hormones (TH), thyroxine (T4) and 3,3′-5′-triiodothyronine (T3) are the necessary and sufficient factors that induce and regulate flatfish metamorphosis. Most of the cellular and molecular action of TH is directed through the binding of T3 to thyroid nuclear receptors bound to promoters with consequent changes in the expression of target genes. The conversion of T4 to T3 and nuclear availability of T3 depends on the expression and activity of a family of 3 selenocysteine deiodinases that activate T4 into T3 or degrade T4 and T3. We have investigated the role of deiodinases in skin and muscle metamorphic changes in halibut. We show that, both at the whole body level and at the cellular level in muscle and skin of the Atlantic halibut ( Hippoglossus hippoglossus ) during metamorphosis, the coordination between activating (D2) and deactivating (D3) deiodinases expression is strongly correlated with the developmental TH-driven changes. The expression pattern of D2 and D3 in cells of both skin and muscle indicate that TH are necessary for the maintenance of larval metamorphic development and juvenile cell types in these tissues. No break in symmetry occurs in the expression of deiodinases and in metamorphic developmental changes occurring both in trunk skin and muscle. The findings that two of the major tissues in both larvae and juveniles maintain their symmetry throughout metamorphosis suggest that the asymmetric changes occurring during flatfish metamorphosis are restricted to the eye and head region. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1473-x Authors M. A. Campinho, Comparative and Molecular Endocrinology Group, Centre for Marine Sciences (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal M. Galay-Burgos, ECETOC, European Centre for Ecotoxicology and Toxicology of Chemicals, 4 avenue E. Van Nieuwenhuyse, B-1160 Brussels, Belgium N. Silva, Comparative and Molecular Endocrinology Group, Centre for Marine Sciences (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal R. A. Costa, Comparative and Molecular Endocrinology Group, Centre for Marine Sciences (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal R. N. Alves, Comparative and Molecular Endocrinology Group, Centre for Marine Sciences (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal G. E. Sweeney, School of Biosciences, Cardiff University, Cardiff, Wales, UK D. M. Power, Comparative and Molecular Endocrinology Group, Centre for Marine Sciences (CCMAR), Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Daily oscillations of clock genes have recently been demonstrated in the ovaries of several species. Clock gene knockout or mutant mice demonstrate a variety of reproductive defects. Accumulating evidence suggests that these rhythms act to synchronise the expression of specific ovarian genes to hypothalamo-pituitary signals and that they are regulated by one or both of the gonadotropins. The aim of this study has been to examine the spatio-temporal expression of the clock genes Per1 and Bmal1 during gonadotropin-independent and gonadotropin-dependent follicle development in the rat ovary. We have examined the ovaries of prepubertal rats, of prepubertal rats stimulated with equine chorionic gonadotropin (eCG)/human chorionic gonadotropin (hCG) and of hypophysectomised adult animals. Using quantitative reverse transcription with the polymerase chain reaction, in situ hybridisation histochemistry and immunohistochemistry, we have demonstrated that the expression of the two clock genes is low and arrhythmic in ovarian cells during early gonadotropin-independent follicle development in prepubertal animals and in hypophysectomised animals. We have also demonstrated that the expression of the clock genes becomes rhythmic following eCG stimulation in the theca interna cells and the secondary interstitial cells and that, following additional hCG stimulation, the expression of the clock genes also becomes rhythmic in the granulosa cells of preovulatory follicles. These findings link the initiation of clock gene rhythms in the rat ovary to the luteinising hormone receptor and suggest a functional link to androgen and progesterone production. In hypophysectomised animals, rhythmic clock gene expression is also observed in the corpora lutea and in secondary interstitial cells demonstrating that, in these compartments, entrainment of clock gene rhythms is gonadotropin-independent. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1489-2 Authors Søren Gräs, Department of Obstetrics and Gynecology, Herlev Hospital, Faculty of Health Sciences, University of Copenhagen, 2730 Herlev, Denmark Birgitte Georg, Depatment of Clinical Biochemistry, Faculty of Health Sciences, Bispebjerg Hospital, University of Copenhagen, 2400 Copenhagen NV, Denmark Henrik L. Jørgensen, Depatment of Clinical Biochemistry, Faculty of Health Sciences, Bispebjerg Hospital, University of Copenhagen, 2400 Copenhagen NV, Denmark Jan Fahrenkrug, Depatment of Clinical Biochemistry, Faculty of Health Sciences, Bispebjerg Hospital, University of Copenhagen, 2400 Copenhagen NV, Denmark Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Cells of multicellular organisms need to communicate with each other and have evolved various mechanisms for this purpose, the most direct and quickest of which is through channels that directly connect the cytoplasms of adjacent cells. Such intercellular channels span the two plasma membranes and the intercellular space and result from the docking of two hemichannels. These channels are densely packed into plasma-membrane spatial microdomains termed “gap junctions” and allow cells to exchange ions and small molecules directly. A hemichannel is a hexameric torus of junctional proteins around an aqueous pore. Vertebrates express two families of gap-junction proteins: the well-characterized connexins and the more recently discovered pannexins, the latter being related to invertebrate innexins (“invertebrate connexins”). Some gap-junctional hemichannels also appear to mediate cell-extracellular communication. Communicating junctions play crucial roles in the maintenance of homeostasis, morphogenesis, cell differentiation and growth control in metazoans. Gap-junctional channels are not passive conduits, as previously long regarded, but use “gating” mechanisms to open and close the central pore in response to biological stimuli (e.g. a change in the transjunctional voltage). Their permeability is finely tuned by complex mechanisms that have just begun to be identified. Given their ubiquity and diversity, gap junctions play crucial roles in a plethora of functions and their dysfunctions are involved in a wide range of diseases. However, the exact mechanisms involved remain poorly understood. Content Type Journal Article Category Review Pages 1-11 DOI 10.1007/s00441-012-1485-6 Authors Jean-Claude Hervé, Institut de Physiologie et Biologie Cellulaires, FRE3511, Université de Poitiers, CNRS, 1 Rue Georges Bonnet, Poitiers, 86022 France Mickaël Derangeon, INSERM, UMR 1087, l’Institut du Thorax, Nantes, France Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Cell block sections serve as an important diagnostic annex for cytological smears, liquid-based SurePath cytology and the Liquid-based Thin-prep Cytology Test (TCT). A variety of methods for the preparation of cell blocks are described in the literature and the techniques in cell blocks are in continuous improvement. A new technique for making cell blocks was introduced in the present study. We first used pregelatinized starch as the frame for the cell block, which is a really simple and economic method, because it can be carried out at room temperature without additional special instruments. We have performed hematoxylin and eosin (HE) staining, immunohistochemistry analysis and fluorescence in situ hybridization (FISH) in the cell block sections in 122 cytological specimens. The results demonstrated in this article show that pregelatinized starch is a useful frame for cell blocks. The pregelatinized starch can effectively collect even a few cells with powerful adhesiveness. Therefore, this new technique for making cell blocks is especially useful for cytologic samples with low cellularity, such as cerebrospinal fluid specimens. Content Type Journal Article Category Regular Article Pages 1-6 DOI 10.1007/s00441-012-1467-8 Authors Qing-Lian He, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Ya-Zhen Zhu, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Guang-Juan Zheng, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Ling-Chun Shi, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Shao-Wei Hu, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Chu-Tian Li, Department of Pathology, Guangdong Provincial Hospital of TCM, Guangzhou University of Chinese Medicine, 111 Dade Road, Guangzhou, 510120 China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Recent investigation of the intestine following ischemia and reperfusion (I/R) has revealed that nitric oxide synthase (NOS) neurons are more strongly affected than other neuron types. This implies that NO originating from NOS neurons contributes to neuronal damage. However, there is also evidence of the neuroprotective effects of NO. In this study, we compared the effects of I/R on the intestines of neuronal NOS knockout (nNOS −/− ) mice and wild-type mice. I/R caused histological damage to the mucosa and muscle and infiltration of neutrophils into the external muscle layers. Damage to the mucosa and muscle was more severe and greater infiltration by neutrophils occurred in the first 24 h in nNOS −/− mice. Immunohistochemistry for the contractile protein, α-smooth muscle actin, was used to evaluate muscle damage. Smooth muscle actin occurred in the majority of smooth muscle cells in the external musculature of normal mice but was absent from most cells and was reduced in the cytoplasm of other cells following I/R. The loss was greater in nNOS −/− mice. Basal contractile activity of the longitudinal muscle and contractile responses to nerve stimulation or a muscarinic agonist were reduced in regions subjected to I/R and the effects were greater in nNOS −/− mice. Reductions in responsiveness also occurred in regions of operated mice not subjected to I/R. This is attributed to post-operative ileus that is not significantly affected by knockout of nNOS. The results indicate that deleterious effects are greater in regions subjected to I/R in mice lacking nNOS compared with normal mice, implying that NO produced by nNOS has protective effects that outweigh any damaging effect of this free radical produced by enteric neurons. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1451-3 Authors Leni R. Rivera, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Louise Pontell, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Hyun-Jung Cho, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Patricia Castelucci, Department of Anatomy, University of São Paulo, São Paulo, Brazil Michelle Thacker, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Daniel P. Poole, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Tony Frugier, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia John B. Furness, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The mdx (X chromosome-linked muscular dystrophy) mouse develops a multi-staged disorder characterized by muscle degeneration and reactive fibrosis. Skeletal muscles of mdx mice are not equally susceptible to degeneration. The aim of this study was to verify whether the intense remodeling of the mdx diaphragm could be attributed to influences from the peritoneal microenvironment and omentum, a lymphohematopoietic tissue rich in progenitor cells and trophic factors. At ages corresponding to increased muscular regeneration (12 weeks) and activation of fibrosis (24 weeks), the mdx omentum exhibited (1) morphological and functional characteristics of activation with enlarged milk-spots, an accumulation of CD4 + , CD8 + and CD19 + B220 + B lymphocytes; (2) the formation of clusters positive for proliferating cell nuclear antigen, mainly in B220 + -rich areas organized in a follicular structure with a germinative center without any challenge by external antigen inducers; (3) clusters with cells positive for fibroblast growth factor-2, numerous Sca-1 + CD3 - CD19 - Mac-1 - progenitor cells and increased CD4 + , CD8 + and CD3 + NK1.1 + cells in the peritoneal cavity. Omentectomy reduced areas with F4/80 + inflammatory infiltrate the activity of matrix metalloproteases 9 and 2, collagen deposition and areas with regenerating myofibers in the diaphragm. Thus, persistent activation of the omentum influences the pattern of inflammation and regeneration of the mdx diaphragm partly via the activation of progenitor cells and the production of growth factors that influence the physiopathology of the muscular tissue remodeling. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1443-3 Authors Douglas Florindo Pinheiro, Department of Immunobiology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Rafael F. da Silva, Department of Cellular and Molecular Biology, Institute of Biology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Lian P. Carvalho, Department of Cellular and Molecular Biology, Institute of Biology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Eustaquio L. Paiva-Oliveira, Department of Immunobiology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Ricardo S. Pereira, Department of Cellular and Molecular Biology, Institute of Biology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Paulo Emilio C. Leite, Department of Cellular and Molecular Biology, Institute of Biology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Maria de Fátima Pinho, Department of Immunobiology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Thereza Quirico-Santos, Department of Cellular and Molecular Biology, Institute of Biology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Jussara Lagrota-Candido, Department of Immunobiology, Fluminense Federal University, Niterói, RJ 24 020 141, Brazil Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Sensory transduction in the cochlea depends on perilymphatic-endolymphatic potassium (K + ) recycling. It has been suggested that the epithelial supporting cells (SCs) of the cochlear duct may form the intracellular K + recycling pathway. Thus, they must be endowed with molecular mechanisms that facilitate K + uptake and release, along with concomitant osmotically driven water movements. As yet, no molecules have been described that would allow for volume-equilibrated transepithelial K + fluxes across the SCs. This study describes the subcellular co-localisation of the K ir 4.1 K + channel (K ir 4.1) and the aquaporin-4 water channel (AQP4) in SCs, on the basis of immunohistochemical double-labelling experiments in rat and human cochleae. The results of this study reveal the expression of K ir 4.1 in the basal or basolateral membranes of the SCs in the sensory domain of the organ of Corti that are adjacent to hair cells and in the non-sensory domains of the inner and outer sulci that abut large extracellular fluid spaces. The SCs of the inner sulcus (interdental cells, inner sulcus cells) and the outer sulcus (Hensen’s cells, outer sulcus cells) display the co-localisation of K ir 4.1 and AQP4 expression. However, the SCs in the sensory domain of the organ of Corti reveal a gap in the expression of AQP4. The outer pillar cell is devoid of both K ir 4.1 and AQP4. The subcellular co-localisation of K ir 4.1 and AQP4 in the SCs of the cochlea described in this study resembles that of the astroglia of the central nervous system and the glial Mueller cells in the retina. Content Type Journal Article Category Regular Article Pages 1-17 DOI 10.1007/s00441-012-1456-y Authors Andreas Eckhard, Hearing Research Centre Department of Otorhinolaryngology-Head & Neck Surgery, University of Tübingen Medical Center, Elfriede-Aulhorn-Straße 5, 72076 Tübingen, Germany Corinna Gleiser, Department of Anatomy, University of Tübingen, Tübingen, Germany Helge Rask-Andersen, Department of Surgical Sciences, Section of Otolaryngology, Uppsala University Hospital, Uppsala, Sweden Heinz Arnold, Hearing Research Centre Department of Otorhinolaryngology-Head & Neck Surgery, University of Tübingen Medical Center, Elfriede-Aulhorn-Straße 5, 72076 Tübingen, Germany Wei Liu, Department of Surgical Sciences, Section of Otolaryngology, Uppsala University Hospital, Uppsala, Sweden Andreas Mack, Department of Anatomy, University of Tübingen, Tübingen, Germany Marcus Müller, Hearing Research Centre Department of Otorhinolaryngology-Head & Neck Surgery, University of Tübingen Medical Center, Elfriede-Aulhorn-Straße 5, 72076 Tübingen, Germany Hubert Löwenheim, Hearing Research Centre Department of Otorhinolaryngology-Head & Neck Surgery, University of Tübingen Medical Center, Elfriede-Aulhorn-Straße 5, 72076 Tübingen, Germany Bernhard Hirt, Hearing Research Centre Department of Otorhinolaryngology-Head & Neck Surgery, University of Tübingen Medical Center, Elfriede-Aulhorn-Straße 5, 72076 Tübingen, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Estrogens diversely affect various physiological processes by genomic or non-genomic mechanisms, in both excitable and non-excitable cells. Additional to the trophic effects of estrogens promoting cell growth and differentiation, recent experimental evidence highlights their involvement in the regulation of intracellular Ca 2+ homeostasis. The effects of estrogens on excitable cells are well documented. However, these steroids also influence numerous physiological events in non-excitable cells, such as fibroblasts or vascular endothelial cells. We have focused our attention on an immortalized endothelial-like cell line derived from fetal bovine cerebellum. Estradiol (E 2 ) effects on intracellular Ca 2+ homeostasis were tested by varying the exposure time to the hormone (8, 24, 48 h). Calcium measurements were performed with genetically encoded Ca 2+ probes (Cameleons) targeted to the main subcellular compartments involved in intracellular Ca 2+ homeostasis (cytosol, endoplasmic reticulum, mitochondria). Mitochondrial Ca 2+ uptake significantly decreased after 48-h exposure to E 2 , whereas cytosolic and endoplasmic reticulum responses were unaffected. The effect of E 2 on mitochondrial Ca 2+ handling was blocked by ICI 182,780, a pure estrogen receptor antagonist, suggesting that the effect was estrogen-receptor–mediated. To evaluate whether the decrease of Ca 2+ uptake affected mitochondrial membrane potential (ΔΨm), cells were monitored in the presence of tetra-methyl-rhodamine-methylester; no significant changes were seen between cells treated with E 2 and controls. To investigate a mechanism of action, we assessed the possibile involvement of the permeability transition pore (PTP), an inner mitochondrial membrane channel influencing energy metabolism and cell viability. We treated cells with CyclosporinA (CsA), which binds to the matrix chaperone cyclophilin-D and regulates PTP opening. CsA reversed the effects of a 48-h treatment with E 2 , suggesting a possible transcriptional modulation of proteins involved in the mitochondrial permeability transition process. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1460-2 Authors Matteo Suman, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Marta Giacomello, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Livio Corain, Department of Management and Engineering, University of Padova, Via S. Nicola 3, 36100 Vicenza (VI), Italy Cristina Ballarin, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Stefano Montelli, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Bruno Cozzi, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Antonella Peruffo, Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell’Università 16, 35020 Legnaro–Agripolis (PD), Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Despite the discovery of the widely expressed CLCA (chloride channel regulators, calcium-activated) proteins more than 15 years ago, their seemingly diverse functions are still poorly understood. With the recent generation of porcine animal models for cystic fibrosis (CF), members of the porcine CLCA family are becoming of interest as possible modulators of the disease in the pig. Here, we characterize pCLCA2, the porcine ortholog of the human hCLCA2 and the murine mCLCA5, which are the only CLCA members expressed in the skin. Immunohistochemical studies with a specific antibody against pCLCA2 have revealed a highly restricted pCLCA2 protein expression in the skin. The protein is strictly co-localized with filaggrin and trichohyalin in the granular layer of the epidermis and the inner root sheath of the hair follicles, respectively. No differences have been observed between the expression patterns of wild-type pigs and CF transmembrane conductance regulator -/- pigs. We speculate that pCLCA2 plays an as yet undefined role in the structural integrity of the skin or, possibly, in specialized functions of the epidermis, including barrier or defense mechanisms. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1482-9 Authors Stephanie Plog, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany Lars Mundhenk, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany Lutz Langbein, German Cancer Research Center, Genetics of Skin Carcinogenesis, A110, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany Achim D. Gruber, Department of Veterinary Pathology, Faculty of Veterinary Medicine, Freie Universität Berlin, Robert-von-Ostertag-Straße 15, 14163 Berlin, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    p53 family members have been implicated in regulation of genomic integrity and apoptosis in a variety of tissues. The Drosophila family member, Dmp53, primarily functions to regulate apoptosis in developing and regenerating tissues but loss of function mutants are viable and fertile. Dmp53 exhibits a striking expression pattern in the male germline with high levels found in nuclear bodies in pre-meiotic germ cells. The localisation of Dmp53 to nuclear bodies is dependent upon Dmp53 complexes being able to bind DNA, and although dmp53 mutants do not affect germline stem cell (GSC) maintenance or differentiation, GSCs are sensitive to overexpression of Dmp53 but maturing spermatogonia are not. Dmp53 thus has differential effects depending upon the stage of male germline maturation. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1479-4 Authors Adrian C. Monk, Department of Anatomy and Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Helen E. Abud, Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia Gary R. Hime, Department of Anatomy and Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Previous studies have shown that craniofacial bone marrow stromal cells (BMSCs) have a strong osteogenic potential. However, the mechanism by which BMSCs of various embryonic origins develop diverse osteogenic potentials remains unclear. To investigate the mechanisms regulating osteoblast differentiation in two different types of BMSCs, we compared the temporal and spatial mRNA and protein expression patterns of Satb2 and its downstream gene Hoxa2 by using real-time polymerase chain reaction, Western blotting and fluorescent immunostaining in mandible BMSCs (M-BMSCs) and tibia BMSCs (T-BMSCs) undergoing osteoblast differentiation. Higher levels of alkaline phosphatase, greater calcium accumulation and earlier expression of Runx2 were observed in osteogenic-induced M-BMSCs compared with T-BMSCs. Low levels of Satb2 were detected in both types of uninduced BMSCs but the majority of SATB2 was located in the nuclei of M-BMSCs. Notably, Satb2 was expressed earlier in M-BMSCs and Hoxa2 , a downstream target of Satb2 , was not expressed in uninduced M-BMSCs or during osteoblast differentiation, just as during embryonic mandible development. In contrast, Hoxa2 was reactivated in T-BMSCs during osteoblast differentiation. Based on these results, we conclude that SATB2 plays a different role during osteoblast differentiation of M-BMSCs and T-BMSCs. The earlier activation of Satb2 expression in M-BMSCs compared with T-BMSCs might explain the stronger osteogenic potential of M-BMSCs. Content Type Journal Article Category Regular Article Pages 1-13 DOI 10.1007/s00441-012-1487-4 Authors Ping Zhang, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Jie Men, Department of Prosthodontics, Shanxi Red Cross Stomatology hospital, 18 Xinghualing, Taiyuan, 030001 Shanxi Province, People’s Republic of China Yu Fu, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Tengfei Shan, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Jinhai Ye, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Yunong Wu, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Zhenjiang Tao, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Laikui Liu, Institute of Stomatology, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Hongbing Jiang, Department of Oral and Maxillofacial Surgery, School of Stomatology, Nanjing Medical University, 136 Hanzhong Road, Nanjing, 210029 Jiangsu Province, People’s Republic of China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The growth/differentiation factor-15, GDF-15, has been found to be secreted by Schwann cells in the lesioned peripheral nervous system. To investigate whether GDF-15 plays a role in peripheral nerve regeneration, we substituted exogenous GDF-15 into 10-mm sciatic nerve gaps in adult rats and compared functional and morphological regeneration to a vehicle control group. Over a period of 11 weeks, multiple functional assessments, including evaluation of pinch reflexes, the Static Sciatic Index and of electrophysiological parameters, were performed. Regenerated nerves were then morphometrically analyzed for the number and quality of regenerated myelinated axons. Substitution of GDF-15 significantly accelerated sensory recovery while the effects on motor recovery were less strong. Although the number of regenerated myelinated axons was significantly reduced after GDF-15 treatment, the regenerated axons displayed advanced maturation corroborating the results of the functional assessments. Our results suggest that GDF-15 is involved in the complex orchestration of peripheral nerve regeneration after lesion. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1493-6 Authors Leonore Mensching, Institute of Neuroanatomy, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany Ann-Kathrin Börger, Institute of Neuroanatomy, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany Xialong Wang, Department of Molecular Embryology, University of Freiburg, Albertstr. 17, 79104 Freiburg, Germany Petar Charalambous, Department of Molecular Embryology, University of Freiburg, Albertstr. 17, 79104 Freiburg, Germany Klaus Unsicker, Department of Molecular Embryology, University of Freiburg, Albertstr. 17, 79104 Freiburg, Germany Kirsten Haastert-Talini, Institute of Neuroanatomy, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Establishing vascularization is a critical obstacle to the generation of engineered heart tissue (EHT) of substantial thickness. Addition of endothelial cells to the formative stages of EHT has been demonstrated to result in prevascularization, or the formation of capillary-like structures. The detailed study of the effects of prevascularization on EHT contractile function is lacking. Here, we evaluated the functional impact of prevascularization by human umbilical vein endothelial cells (HUVECs) in self-organizing EHT. EHT fibers were generated by the self-organization of neonatal rat cardiac cells on a fibrin hydrogel scaffold with or without HUVECs. Contractile function was measured and force–length relationship and rate of force production were assessed. Immunofluorescent studies were used to evaluate arrangement and distribution of HUVECs within the EHT fibers. RT-PCR was used to assess the transcript levels of hypoxia inducible factor-1a (Hif-1α). EHT with HUVECs manifested tubule-like structures at the periphery during fiber formation. After fiber formation, HUVECs were heterogeneously located throughout the EHT fiber and human CD31+ tubule-like structures were identified. The expression level of Hif-1α did not change with the addition of HUVECs. However, maximal force and rate of force generation were not improved in HUVECs containing EHT as compared to control EHT fibers. The addition of HUVECs may result in sparse microvascularization of EHT. However, this perceived benefit is overshadowed by a significant decrease in contractile function and highlights the need for perfused vascularization strategies in order to generate EHT that approaches clinically relevant dimensions. Content Type Journal Article Category Short Communication Pages 1-6 DOI 10.1007/s00441-012-1492-7 Authors Claus Svane Sondergaard, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Russell Witt, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Grant Mathews, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Skender Najibi, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Lisa Le, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Tracy Clift, Department of Cardiothoracic Surgery, University of California at Davis Medical Center, Sacramento, CA 95817, USA Ming-Sing Si, Institute for Regenerative Cures, University of California at Davis Medical Center, Sacramento, CA, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The response of insect olfactory receptor neurons (ORNs) involves an increase in intracellular Ca 2+ concentration, as in vertebrate ORNs. In order to decipher the Ca 2+ clearance mechanisms in insect ORNs, we have investigated the presence of a plasma membrane Ca 2+ ATPase (PMCA) in the peripheral olfactory system of the moth Spodoptera littoralis . From an analysis of a male antennal expressed-sequence-tag database combined with a strategy of 5′/3′ rapid amplification of cDNA ends plus the polymerase chain reaction, we have cloned a full-length cDNA encoding a PMCA. In adult males, the PMCA transcript has been found in various tissues, including the antennae in which its presence has been detected in the sensilla trichodea , and in cultured ORNs. The PMCA gene is slightly expressed at the end of the pupal stage, reaches a maximum at emergence and is maintained at a high level during the adult period. Taken together, these results provide, for the first time, molecular evidence for the putative participation of a PMCA in signalling pathways responsible for the establishment and functioning of the insect peripheral olfactory system. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1483-8 Authors Adrien François, UPMC - Université Paris 6, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, 7 Quai Saint-Bernard, 75252 Paris cedex 05, France Françoise Bozzolan, UPMC - Université Paris 6, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, 7 Quai Saint-Bernard, 75252 Paris cedex 05, France Elodie Demondion, INRA, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, Route de Saint-Cyr, 78026 Versailles cedex, France Nicolas Montagné, UPMC - Université Paris 6, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, 7 Quai Saint-Bernard, 75252 Paris cedex 05, France Philippe Lucas, INRA, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, Route de Saint-Cyr, 78026 Versailles cedex, France Stéphane Debernard, UPMC - Université Paris 6, UMR-A 1272 Physiologie de l’Insecte: Signalisation et Communication, 7 Quai Saint-Bernard, 75252 Paris cedex 05, France Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The clinical translation of tissue engineering approaches is limited by the requirement of a cell source. Cell guidance is a new concept that provides an alternative approach, obviating a requirement for an external cell source. This relies on site-specific homing and differentiation of the patient’s own cells to an implanted scaffold through controlled delivery of cytokines. In this study, we used stromal-cell-derived factor 1-alpha (SDF-1α) in combination with bone morphogenic protein (BMP)-2 or transforming growth factor (TGF)-β1 to induce cell migration and osteogenic or chondrogenic differentiation, respectively, in implanted scaffolds in a rat model. A customized cytokine microdelivery apparatus was used to ensure the constant rate and concentration of cytokine delivery around the scaffold. The formation of osteoid or early cartilage was observed after 4 weeks in specimens treated with SDF-1α and either BMP-2 or TGF-β1. The density of cellular infiltrate and formation of differentiated tissue were lower in scaffolds treated only with BMP-2 or TGF-β1. Thus, controlled SDF-1α delivery induces cell migration into scaffolds and can result in enhanced osteogenesis and chondrogenesis when used in combination with differentiation cytokines for purposes of tissue engineering. Content Type Journal Article Category Short Communication Pages 1-6 DOI 10.1007/s00441-012-1449-x Authors Harvey Chim, Department of Plastic Surgery, Case Western Reserve University, 11100 Euclid Avenue, Cleveland, OH 44106 USA Erin Miller, Department of Plastic Surgery, Case Western Reserve University, 11100 Euclid Avenue, Cleveland, OH 44106 USA Christy Gliniak, Department of Plastic Surgery, Case Western Reserve University, 11100 Euclid Avenue, Cleveland, OH 44106 USA Eben Alsberg, Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The mitochondrial genome contributes key proteins to the electron-transfer chain, which through oxidative phosphorylation, generates the vast majority of cellular ATP. This maternally inherited genome is transmitted to subsequent generations through the oocyte. Its transmission, inheritance and replication are strictly regulated so that fully mature cells can be appropriately populated with mitochondrial DNA once they mature into adult cells. However, gametes do not always acquire the appropriate numbers of mitochondrial DNA copy; this often renders them inappropriate for successful fertilisation outcome. Furthermore, the number of assisted reproductive technologies that can overcome problems associated with infertility and that can provide enhanced genetic outcomes for the offspring is increasing. However, such techniques could also have a detrimental effect on offspring survival. If we are to introduce these technologies into in vitro fertilisation clinics and animal production, then we first need to validate their use carefully. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-012-1444-2 Authors Justin C. St John, Centre for Reproduction and Development, Monash Institute of Medical Research, 27-31 Wright Street, Clayton, Vic 3168, Australia Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    We describe the structure of the spleen of the African lungfish Protopterus annectens in freshwater conditions, and after 6 months of aestivation. The spleen is formed by cortical tissue that surrounds the splenic parenchyma. The cortex is a reticulum that contains two types of granulocytes, developing and mature plasma cells, and melanomacrophage centres (MMCs). The parenchyma is divided into lobules that show a subcapsular sinus and areas of red pulp and white pulp. Red pulp contains vascular sinuses and atypical cords formed by delicate trabeculae. White pulp also contains vascular sinuses and cords. Structural data indicate that red pulp is involved in erythropoiesis, destruction of effete erythrocytes, and plasma cell differentiation. White pulp appears to be involved in the production of immune responses. Macrophages and sinus endothelial cells constitute the reticulo-endothelial system of the spleen. After aestivation, the number of MMCs increases, and spleen tissue is infiltrated by lymphocytes, granulocytes, and monocytes. Also, white pulp is reduced, and sinus endothelial cells undergo vacuolar degeneration. Lungfish spleen shares structural characteristics with secondary lymphoid organs of both ectothermic and endothermic vertebrates, but appears to have evolved in unique ways. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1462-0 Authors José M. Icardo, Department of Anatomy and Cell Biology, Faculty of Medicine, University of Cantabria, c/ Cardenal Herrera Oria, s/n, 39011 Santander, Spain Wai P. Wong, Department of Biological Sciences, National University of Singapore, Singapore, 117543 Republic of Singapore Elvira Colvee, Department of Anatomy and Cell Biology, Faculty of Medicine, University of Cantabria, c/ Cardenal Herrera Oria, s/n, 39011 Santander, Spain Ai M. Loong, Department of Biological Sciences, National University of Singapore, Singapore, 117543 Republic of Singapore Yuen K. Ip, Department of Biological Sciences, National University of Singapore, Singapore, 117543 Republic of Singapore Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Urocortin (UCN) is a 40-amino-acid peptide and a member of the corticotropin-releasing hormone (CRH) family, which includes CRH, urotensin I, sauvagine, UCN2 and UCN3. The biological actions of CRH family peptides are mediated via two types of G-protein-coupled receptors, namely CRH type 1 receptor (CRHR1) and CRH type 2 receptor (CRHR2). The biological effects of these peptides are mediated and modulated not only by CRH receptors but also via a highly conserved CRH-binding protein (CRHBP). Our aim was to investigate the expression of UCN, CRHR1, CRHR2 and CRHBP by immunohistochemistry, Western blot and reverse transcription with the polymerase chain reaction (RT-PCR) in the horse thyroid gland. The results showed that UCN, CRHR1 and CRHR2 were expressed in the thyroid gland, whereas CRHBP was not expressed. Specifically, UCN immunoreactivity (-IR) was found in the thyroid follicular cells, CRHR2-IR in the C-cells and CRHR1-IR in blood vessels. Western blot analysis and RT-PCR experiments confirmed the immunohistochemical data. These results suggest that a regulatory system exists in the mammalian thyroid gland based on UCN, CRHR1 and CRHR2 and that UCN plays a role in the regulation of thyroid physiological functions through a paracrine mechanism. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1450-4 Authors Caterina Squillacioti, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Adriana De Luca, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Sabrina Alì, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Salvatore Paino, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Giovanna Liguori, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Nicola Mirabella, Department of Structure, Functions and Biological Technologies, University of Naples Federico II, Via Veterinaria 1, 80137 Naples, Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Fertilization of an egg by a spermatozoon sets the stage for mammalian development. Viable sperm are a prerequisite for successful fertilization and beyond. Spermatozoa have a unique cell structure where haploid genomic DNA is located in a tiny cytoplasmic space in the head, mitochondria in the midpiece and then the tail, all enclosed by several layers of membrane. Proteins in sperm play vital roles in motility, capacitation, fertilization, egg activation and embryo development. Molecular defects in these proteins are associated with low fertility or in some cases, infertility. This review will first summarize genesis, molecular anatomy and physiology of spermatozoa, fertilization, embryogenesis and then those proteins playing important roles in various aspects of sperm physiology. Content Type Journal Article Category Review Pages 1-11 DOI 10.1007/s00441-012-1452-2 Authors Aruna Govindaraju, Department of Animal and Dairy Sciences, Mississippi State University, Starkville, 39762 MS, USA Sule Dogan, Department of Animal and Dairy Sciences, Mississippi State University, Starkville, 39762 MS, USA Nelida Rodriguez-Osorio, Grupo Centauro Universidad de Antioquia, Medellin, Colombia Kamilah Grant, Department of Animal and Dairy Sciences, Mississippi State University, Starkville, 39762 MS, USA Abdullah Kaya, Alta Genetics, Inc., Watertown, WI, USA Erdogan Memili, Department of Animal and Dairy Sciences, Mississippi State University, Starkville, 39762 MS, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Although several studies have shown a dysregulation of microRNA (miRNA) expression profiles in cutaneous melanoma, there has been little research on the miRNA machinery itself. In this study, we investigated the mRNA expression profiles of different miRNA machinery components in primary cutaneous malignant melanoma (PCMM), cutaneous malignant melanoma metastases (CMMM) and benign melanocytic nevi (BMN). Patients with PCMM ( n  = 7), CMMM ( n  = 6) and BMN ( n  = 7) were included in the study. Punch biopsies were harvested from the centers of tumors (lesional) and from BMN (control). In contrast to previous reports exploring specific clusters of miRNAs in PCMM, the present study investigates mRNA expression levels of Dicer, Drosha, Exp5, DGCR8 and the RISC components PACT, argonaute-1, argonaute-2, TARBP1, TARBP2, MTDH and SND1, which were detected by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR). Argonaute-1, TARBP2 and SND1 expression levels were significantly higher in BMN compared to PCMM ( p  〈 0.05). TARBP2 expression levels were significantly higher in CMMM compared to PCMM ( p  〈 0.05). SND1 expression levels were significantly higher in CMMM compared to PCMM and BMN ( p  〈 0.05). Dicer, Drosha, DGCR8, Exp5, argonaute-2, PACT, TARBP1 and MTDH expression levels showed no significant differences within groups ( p  〉 0.05). The results of this study show that the miRNA machinery components argonaute-1, TARBP2 and SND1 are dysregulated in PCMM and CMMM compared to BMN and may play a role in the process of malignant transformation. Content Type Journal Article Category Regular Article Pages 1-8 DOI 10.1007/s00441-012-1446-0 Authors Michael Sand, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Marina Skrygan, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Dimitrios Georgas, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Daniel Sand, Department of Medicine, Olive View UCLA Medical Center, University of California (UCLA), Los Angeles, CA 91342, USA Thilo Gambichler, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Peter Altmeyer, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Falk G. Bechara, Department of Dermatology, Venereology and Allergology, Ruhr-University Bochum, St. Josef Hospital, Gudrunstr. 56, 44791 Bochum, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Mutations in the tissue-nonspecific alkaline phosphatase (TNAP) gene can result in skeletal and dental hypomineralization and severe neurological symptoms. TNAP is expressed in the synaptic cleft and the node of Ranvier in normal adults. Using TNAP knockout (KO) mice (Akp2 -/- ), we studied synaptogenesis and myelination with light- and electron microscopy during the early postnatal days. Ablation of TNAP function resulted in a significant decrease of the white matter of the spinal cord accompanied by ultrastructural evidence of cellular degradation around the paranodal regions and a decreased ratio and diameter of the myelinated axons. In the cerebral cortex, myelinated axons, while present in wild-type, were absent in the Akp2 -/- mice and these animals also displayed a significantly increased proportion of immature cortical synapses. The results suggest that TNAP deficiency could contribute to neurological symptoms related to myelin abnormalities and synaptic dysfunction, among which epilepsy, consistently present in the Akp2 -/- mice and observed in severe cases of hypophosphatasia. Content Type Journal Article Category Regular Article Pages 1-13 DOI 10.1007/s00441-012-1455-z Authors János Hanics, Department of Anatomy, Histology and Embryology, Semmelweis University, Budapest, Hungary János Barna, Department of Anatomy, Histology and Embryology, Semmelweis University, Budapest, Hungary Jinsong Xiao, Centre de recherche Cerveau et Cognition (CerCo) UMR5549, CNRS and Université de Toulouse (UPS), Pavillon Baudot, CHU Purpan, BP 25202, 31052 Toulouse Cedex 3, France José Luis Millán, Sanford Children’s Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, CA, USA Caroline Fonta, Centre de recherche Cerveau et Cognition (CerCo) UMR5549, CNRS and Université de Toulouse (UPS), Pavillon Baudot, CHU Purpan, BP 25202, 31052 Toulouse Cedex 3, France László Négyessy, Department of Theory, Institute for Particle and Nuclear Physics, Wigner Research Centre for Physics, Hungarian Academy of Sciences, P.O. Box 49, Budapest, 1525 Hungary Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Reconstructing a functional organ of Corti is the ultimate target towards curing hearing loss. Despite the impressive technical gains made over the last few years, many complications remain ahead for the two main restoration avenues: in vitro transformation of pluripotent cells into hair cell-like cells and adenovirus-mediated gene therapy. Most notably, both approaches require a more complete understanding of the molecular networks that ensure specific cell types form in the correct places to allow proper function of the restored organ of Corti. Important to this understanding are the basic helix-loop-helix (bHLH) transcription factors (TFs) that are highly diverse and serve to increase functional complexity but their evolutionary implementation in the inner ear neurosensory development is less conspicuous. To this end, we review the evolutionary and developmentally dynamic interactions of the three bHLH TFs that have been identified as the main players in neurosensory evolution and development, Neurog1, Neurod1 and Atoh1. These three TFs belong to the neurogenin / atonal family and evolved from a molecular precursor that likely regulated single sensory cell development in the ectoderm of metazoan ancestors but are now also expressed in other parts of the body, including the brain. They interact extensively via intracellular and intercellular cross-regulation to establish the two main neurosensory cell types of the ear, the hair cells and sensory neurons. Furthermore, the level and duration of their expression affect the specification of hair cell subtypes (inner hair cells vs. outer hair cells). We propose that appropriate manipulation of these TFs through their characterized binding sites may offer a solution by itself, or in conjunction with the two other approaches currently pursued by others, to restore the organ of Corti. Content Type Journal Article Category Review Pages 1-18 DOI 10.1007/s00441-012-1454-0 Authors Ning Pan, Department of Biology, University of Iowa, College of Liberal Arts and Sciences, Iowa City, IA 52242, USA Benjamin Kopecky, Department of Biology, University of Iowa, College of Liberal Arts and Sciences, Iowa City, IA 52242, USA Israt Jahan, Department of Biology, University of Iowa, College of Liberal Arts and Sciences, Iowa City, IA 52242, USA Bernd Fritzsch, Department of Biology, University of Iowa, College of Liberal Arts and Sciences, Iowa City, IA 52242, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Within a few decades, the repair of long neuronal pathways such as spinal cord tracts, the optic nerve or intracerebral tracts has gone from being strongly contested to being recognized as a potential clinical challenge. Cut axonal stumps within the optic nerve were originally thought to retract and become irreversibly necrotic within the injury zone. Optic nerve astrocytes were assumed to form a gliotic scar and remodelling of the extracellular matrix to result in a forbidden environment for re-growth of axons. Retrograde signals to the ganglion cell bodies were considered to prevent anabolism, thus also initiating apoptotic death and gliotic repair within the retina. However, increasing evidence suggests the reversibility of these regressive processes, as shown by the analysis of molecular events at the site of injury and within ganglion cells. We review optic nerve repair from the perspective of the proximal axon stump being a major player in determining the successful formation of a growth cone. The axonal stump and consequently the prospective growth cone, communicates with astrocytes, microglial cells and the extracellular matrix via a panoply of molecular tools. We initally highlight these aspects on the basis of recent data from numerous laboratories. Then, we examine the mechanisms by which an injury-induced growth cone can sense its surroundings within the area distal to the injury. Based on requirements for successful axonal elongation within the optic nerve, we explore the models employed to instigate successful growth cone formation by ganglion cell stimulation and optic nerve remodelling, which in turn accelerate growth. Ultimately, with regard to the proteomics of regenerating retinal tissue, we discuss the discovery of isoforms of crystallins, with crystallin beta-b2 (crybb2) being clearly upregulated in the regenerating retina. Crystallins are produced and used to promote the elongation of growth cones. In vivo and in vitro, crystallins beta and gamma additionally promote the growth of axons by enhancing the production of ciliary neurotrophic factor (CNTF), indicating that they also act on astrocytes to promote axonal regrowth synergistically. These are the first data showing that axonal regeneration is related to crybb2 movement within neurons and to additional stimulation of CNTF. We demonstrate that neuronal crystallins constitute a novel class of neurite-promoting factors that probably operate through an autocrine and paracrine mechanism and that they can be used in neurodegenerative diseases. Thus, the post-injury fate of neurons cannot be seen merely as inevitable but, instead, must be regarded as a challenge to shape conditions for initiating growth cone formation to repair the damaged optic nerve. Content Type Journal Article Category Review Pages 1-21 DOI 10.1007/s00441-012-1442-4 Authors Solon Thanos, Institute of Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, D15, 48149 Münster, Germany Michael R. R. Böhm, Institute of Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, D15, 48149 Münster, Germany Maurice Schallenberg, Department of Ophthalmology, University Essen/Duisburg, School of Medicine, Hufelandstrasse 5, Essen, Germany Patrick Oellers, Institute of Experimental Ophthalmology, School of Medicine, University of Münster, Albert-Schweitzer-Campus 1, D15, 48149 Münster, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Pseudoephedrine, an over-the-counter drug, is commonly used for the treatments of asthma, nasal congestion, and obesity. Furthermore, it can be used as a psychostimulant drug if taken in large doses; however, there have been no reports on its effects on reproduction. The aim of this study was therefore to investigate the effects of pseudoephedrine administration on sperm morphology, sperm concentration and apoptotic activity in the rat testis. Rats were administered intraperitoneally (IP) with pseudoephedrine at 120 mg/kg for the acute group and 80 mg/kg, IP, once daily for 15 days for the chronic group, while a control group was treated with vehicle. The percentages of normal sperm morphology were significantly decreased in both acute and chronic groups when compared with controls while the total sperm count was significantly decreased in the acute group. Apoptotic activities were increased significantly in both pseudoephedrine-treated groups. The results indicate that pseudoephedrine can induce sperm abnormalities, decrease sperm numbers and increase apoptotic activity in the testis of rats if taken at high doses. The results of this study suggest that the users of pseudoephedrine in medical treatments need to be aware of its potential toxicity involving spermatogenesis. Content Type Journal Article Category Regular Article Pages 1-6 DOI 10.1007/s00441-012-1408-6 Authors Sutisa Nudmamud-Thanoi, Department of Anatomy, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand Samur Thanoi, Department of Anatomy, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Obesity affects sex hormone secretion, which can negatively influence prostatic structure, homeostasis, and disease. This investigation aimed to evaluate the repercussions of obesity induced by a high-fat diet on the rat prostate, with or without treatment with the aromatase inhibitor, Letrozole. Adult Wistar rats were fed a high-fat diet (20% saturated fat, O) for 15 weeks to induce obesity or received a balanced diet (4% fat, C). Then, a group of C and O rats were daily treated with Letrozole (1 mg/kg b.w. per day) for 2 weeks (CL and OL, respectively). Subsequently, ventral prostate was processed for analysis by transmission electron microscopy, immunohistochemistry, and Western blotting. Obesity decreased 70% of the testosterone plasma level. The prostate showed epithelial atrophy and dilated acini in the intermediate portion and epithelial wrinkling in the distal tips. The relative frequency of smooth muscle α-actin in the O group increased by 67%. Ultrastructurally, epithelial cells in obese animals presented altered secretory organelles, lipid droplets, and thicker subjacent fibromuscular layer. Letrozole treatment caused a partial restoration of the prostatic changes caused by obesity. Obesity increased the prostatic content of fibroblast growth factor-2 (FGF-2) by 150%, and Letrozole treatment increased this protein even more in the control and obese groups. This investigation shows that obesity provokes structural and ultrastructural changes in the epithelium of rat prostate; these changes might affect gland homeostasis and physiology. The epithelial and smooth muscle cell hyperplasia and increased FGF-2 expression observed in this experimental model of obesity/insulin-resistance might explain the high frequency of benign prostatic hyperplasia in insulin-resistant men. Content Type Journal Article Category Regular Article Pages 1-12 DOI 10.1007/s00441-012-1420-x Authors Daniele Lisboa Ribeiro, Department of Histology, Institute of Biomedical Sciences, Federal University of Uberlandia (UFU), Uberlândia, Minas Gerais, Brazil Maria Etelvina Pinto, Department of Biology, Institute of Biosciences, Letters and Exact Sciences, University Estadual Paulista (UNESP), São José do Rio Preto, São Paulo, Brazil Samantha Yuri Maeda, Department of Biology, Institute of Biosciences, Letters and Exact Sciences, University Estadual Paulista (UNESP), São José do Rio Preto, São Paulo, Brazil Sebastião Roberto Taboga, Department of Biology, Institute of Biosciences, Letters and Exact Sciences, University Estadual Paulista (UNESP), São José do Rio Preto, São Paulo, Brazil Rejane Maira Góes, Department of Biology, Institute of Biosciences, Letters and Exact Sciences, University Estadual Paulista (UNESP), São José do Rio Preto, São Paulo, Brazil Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    We compared the two sources of adipose and bone marrow-derived mesenchymal stem cells (BMSCs and AMSCs ) in multiple differentiation capacity and biological characteristics to provide a theoretical basis for stem cells transplantation. We isolated bone marrow- and adipose-derived mesenchymal stem cells and compared their phenotype,cell doubling time, the secretion of factors and their ability of multi-differentiation. We also compared their differences in T lymphocyte activation, proliferation and suppression. BMSCs and AMSCs were similar in cell phenotype and the differences existed only in the expression of CD106. On the proliferation rate, AMSCs were faster than BMSCs (doubling time 28 vs. 39 h). In addition, both of these two sources of cells were able to differentiate into bone, fat and cartilage that proved their stem cells properties and the number of stem cell progenitors (CFU-F) from adipose tissue were 10 times larger than those from bone marrow. But AMSCs showed a diminished capacity for suppressing T lymphocyte proliferation and activation compared to BMSCs. Cell origin and abundance were decisive factors in stem cells applications and, in the same volume, with the same premise of AMSCs and BMSCs, adipose tissue is a more promising source of stem cells. Content Type Journal Article Category Regular Article Pages 1-11 DOI 10.1007/s00441-012-1453-1 Authors Xishan Zhu, Department of Internal Oncology, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038 China Wei Shi, Department of Internal Oncology, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038 China Weiping Tai, Department of Internal Oncology, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038 China Fuquan Liu, Department of Interventional theraphy, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038 People’s Republic of China Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Fish scale culture can be used as a model to test the effects of several molecules on bone metabolism by histological and biochemical methods, although solid cell biology data about the behavior of the scale cells in culture are needed if such a model is to be employed for pharmacological applications. In the present study, we cultured zebrafish scales at various temperatures and for various times and analyzed the behavior of the bone cells in terms of viability and activity. We demonstrated that the cultured scale cells maintained their usual distribution at 28°C until 72 h, after which time episquamal osteoblasts showed an obvious change in their cell organization followed by an increase in cell death. Osteoclast tartrate-resistant acid phosphatase and osteoblast alkaline phosphatase activities were maintained until 72 h but were reduced at 96 h as a consequence of the massive cell death. This scenario indicates that zebrafish scales cultured until 72 h can be considered as an innovative model of explanted organ culture to assay the ability of chemical compounds to modulate the metabolism of bone cells. Content Type Journal Article Category Regular Article Pages 1-7 DOI 10.1007/s00441-012-1436-2 Authors Sara Pasqualetti, Gruppo San Donato Foundation, Milan, Italy Giuseppe Banfi, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy Massimo Mariotti, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description: Special issue Heidelberg Heart II: Abstracts of oral and poster presentations Content Type Journal Article Pages 1-36 DOI 10.1007/s00441-012-1412-x Authors Werner W. Franke, German Cancer Research Center, Im Neuenheimer Feld 280, Heidelberg, 69120 Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Functional studies have shown that subsets of autonomic preganglionic neurons respond to ghrelin and ghrelin mimetics and in situ hybridisation has revealed receptor gene expression in the cell bodies of some preganglionic neurons. Our present goal has been to determine which preganglionic neurons express ghrelin receptors by using mice expressing enhanced green fluorescent protein (EGFP) under the control of the promoter for the ghrelin receptor (also called growth hormone secretagogue receptor). The retrograde tracer Fast Blue was injected into target organs of reporter mice under anaesthesia to identify specific functional subsets of postganglionic sympathetic neurons. Cryo-sections were immunohistochemically stained by using anti-EGFP and antibodies to neuronal markers. EGFP was detected in nerve terminal varicosities in all sympathetic chain, prevertebral and pelvic ganglia and in the adrenal medulla. Non-varicose fibres associated with the ganglia were also immunoreactive. No postganglionic cell bodies contained EGFP. In sympathetic chain ganglia, most neurons were surrounded by EGFP-positive terminals. In the stellate ganglion, neurons with choline acetyltransferase immunoreactivity, some being sudomotor neurons, lacked surrounding ghrelin-receptor-expressing terminals, although these terminals were found around other neurons. In the superior cervical ganglion, the ghrelin receptor terminals innervated subgroups of neurons including neuropeptide Y (NPY)-immunoreactive neurons that projected to the anterior chamber of the eye. However, large NPY-negative neurons projecting to the acini of the submaxillary gland were not innervated by EGFP-positive varicosities. In the celiaco-superior mesenteric ganglion, almost all neurons were surrounded by positive terminals but the VIP-immunoreactive terminals of intestinofugal neurons were EGFP-negative. The pelvic ganglia contained groups of neurons without ghrelin receptor terminal innervation and other groups with positive terminals around them. Ghrelin receptors are therefore expressed by subgroups of preganglionic neurons, including those of vasoconstrictor pathways and of pathways controlling gut function, but are absent from some other neurons, including those innervating sweat glands and the secretomotor neurons that supply the submaxillary salivary glands. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1405-9 Authors John B. Furness, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Hyun-Jung Cho, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Billie Hunne, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Haruko Hirayama, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Brid P. Callaghan, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Alan E. Lomax, Gastrointestinal Diseases Research Unit, Queen’s University, Kingston, Ontario, Canada James A. Brock, Department of Anatomy & Neuroscience, University of Melbourne, Parkville, Victoria 3010, Australia Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    This study provides a new perspective on the long-standing problem of the nature of the decapod crustacean blood-brain interface. Previous studies of crustacean blood-brain interface permeability have relied on invasive histological, immunohistochemical and electrophysiological techniques, indicating a leaky non-selective blood-brain barrier. The present investigation involves the use of magnetic resonance imaging (MRI), a method for non-invasive longitudinal tracking of tracers in real-time. Differential uptake rates of two molecularly distinct MRI contrast agents, namely manganese (Mn(II)) and Magnevist® (Gd-DTPA), were observed and quantified in the crayfish, Cherax destructor . Contrast agents were injected into the pericardium and uptake was observed with longitudinal MRI for approximately 14.5 h. Mn(II) was taken up quickly into neural tissue (within 6.5 min), whereas Gd-DTPA was not taken up into neural tissue and was instead restricted to the intracerebral vasculature or excreted into nearby sinuses. Our results provide evidence for a charge-selective intracerebral blood-brain interface in the crustacean nervous system, a structural characteristic once considered too complex for a lower-order arthropod. Content Type Journal Article Category Regular Article Pages 1-11 DOI 10.1007/s00441-012-1413-9 Authors Adriane G. Otopalik, Neuroscience Program, Wellesley College, Wellesley, MA 02481 USA Jane Shin, Chemistry Department, Wellesley College, 106 Central Street, Wellesley, MA 02481 USA Barbara S. Beltz, Neuroscience Program, Wellesley College, Wellesley, MA 02481 USA David C. Sandeman, Neuroscience Program, Wellesley College, Wellesley, MA 02481 USA Nancy H. Kolodny, Chemistry Department, Wellesley College, 106 Central Street, Wellesley, MA 02481 USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    UCH-L1 (ubiquitin carboxyl terminal hydrolase L1) is well known as an enzyme that hydrolyzes polyubiquitin at its C-terminal to release ubiquitin monomers. Although the overexpression of UCH-L1 inhibits proteasome activity in cultured cells, its biological significance in living organisms has not been clarified in detail. Here, we utilized Drosophila as a model system to examine the effects of the overexpression of dUCH, a Drosophila homologue of UCH-L1, on development. Overexpression in the eye imaginal discs induced a rough eye phenotype in the adult, at least partly resulting from the induction of caspase-dependent apoptosis followed by compensatory proliferation. Genetic crosses with enhancer trap lines marking the photoreceptor cells also revealed that the overexpression of dUCH specifically impaired R7 photoreceptor cell differentiation with a reduction in activated extracellular-signal-regulated kinase signals. Furthermore, the dUCH-induced rough eye phenotype was rescued by co-expression of the sevenless gene or the Draf gene, a downstream component of the mitogen-activated protein kinase (MAPK) cascade. These results indicate that the overexpression of dUCH impairs R7 photoreceptor cell differentiation by down-regulating the MAPK pathway. Interestingly, this process appears to be independent of its pro-apoptotic function. Content Type Journal Article Category Regular Article Pages 1-11 DOI 10.1007/s00441-012-1404-x Authors Dang Thi Phuong Thao, Department of Molecular and Environmental Biotechnology, University of Science, Vietnam National University in Ho Chi Minh City, Ho Chi Minh City, Vietnam Phan Nguyen Thuy An, Department of Molecular and Environmental Biotechnology, University of Science, Vietnam National University in Ho Chi Minh City, Ho Chi Minh City, Vietnam Masamitsu Yamaguchi, Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Kyoto, Japan Tran LinhThuoc, Department of Molecular and Environmental Biotechnology, University of Science, Vietnam National University in Ho Chi Minh City, Ho Chi Minh City, Vietnam Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Evidence has emerged recently indicating that differentiation is not entirely a one-way process, and that it is possible to convert one cell type to another, both in vitro and in vivo. This phenomenon is called transdifferentiation, and is generally defined as the stable switch of one cell type to another. Transdifferentiation plays critical roles during development and in regeneration pathways in nature. Although this phenomenon occurs rarely in nature, recent studies have been focused on transdifferentiation and the reprogramming ability of cells to produce specific cells with new phenotypes for use in cell therapy and regenerative medicine. Thus, understanding the principles and the mechanism of this process is important for producing desired cell types. Here some well-documented examples of transdifferentiation, and their significance in development and regeneration are reviewed. In addition, transdifferentiation pathways are considered and their potential molecular mechanisms, especially the role of master switch genes, are considered. Finally, the significance of transdifferentiation in regenerative medicine is discussed. Content Type Journal Article Category Review Pages 1-18 DOI 10.1007/s00441-012-1403-y Authors Sajjad Sisakhtnezhad, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran P.O. Box: 9177948974 Maryam M. Matin, Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran P.O. Box: 9177948974 Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The bursa of Fabricius of the chicken is known as a primary lymphoid organ for B-cell development. Morphologically, the origin of IgG-containing cells in the bursa has not been clear until now, because abundant maternal IgG (MIgG) is transported to the chick embryo and distributed to the bursal tissue around hatching. Thus, it has been difficult to find out whether these cells themselves biosynthesize IgG or if they acquire MIgG via attachment to their surface. Our present study employing in situ hybridization clarified that IgG-containing cells in the medulla of bursal follicles did not biosynthesize IgG. To study the role of MIgG in the development of those IgG-containing cells, MIgG-free chicks were established from surgically bursectomized hen (SBx-hen). We found that, on the one hand, deprivation of MIgG from chicks completely inhibited the development of IgG-containing cells in the medulla after hatching. On the other hand, administration of MIgG to MIgG-free chicks recovered the emergence of those cells. In addition, we observed that those cells did not bear a B-cell marker and possessed dendrites with aggregated IgG. These results demonstrate that IgG-containing cells in the medulla are reticular cells that capture aggregated MIgG. Moreover, we show that the isolation of the bursa from environmental stimuli by bursal duct ligation (BDL) suppressed the development of IgG-containing cells after hatching. Thus, it is implied that environmental stimulations play a key role in MIgG aggregations and dendritic distributions of aggregated MIgG in the medulla after hatching. Content Type Journal Article Category Regular Article Pages 1-14 DOI 10.1007/s00441-012-1407-7 Authors Shigeo Ekino, Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Honjo, Kumamoto, Japan Hiroshi Arakawa, Helmholtz Center Munich, German Research Center for Environmental Health, Institute for Molecular Radiobiology and Institute for Radiocytogenetics, Neuherberg, Germany Kayoko Sonoda, Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Honjo, Kumamoto, Japan Kazuhiro Noguchi, Department of Histology, Graduate School of Medical Sciences, Kumamoto University, Honjo, Kumamoto, Japan Seiji Inui, Department of Immunology and Hematology, School of Health Sciences, Kumamoto University, Kuhonji, Kumamoto, Japan Hideaki Yokoyama, EW Nutrition Japan K.K., Sano, Gifu, Japan Yoshikatsu Kodama, EW Nutrition Japan K.K., Sano, Gifu, Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Studies carried out during the last 2 decades have dramatically increased our knowledge of the pathways and mechanisms of intracellular membrane traffic, most recently due to the developments in light microscopy and in vivo imaging of fluorescent fusion proteins. These studies have also revealed that certain molecules do not behave according to the classical transportation rules first documented in cell biology textbooks in the 1980s and 1990s. Initially, unconventional mechanisms of secretion that do not involve passage of cargo through the stacked Golgi cisternae were thought to confer on cells the ability to discard excess amounts of protein products. With time, however, more physiological mechanisms and roles have been proposed for an increasing number of secretory processes that bypass the Golgi apparatus. Content Type Journal Article Category Review Pages 1-16 DOI 10.1007/s00441-012-1409-5 Authors Kristian Prydz, Department of Molecular Biosciences, University of Oslo, Oslo, Norway Heidi Tveit, Department of Molecular Biosciences, University of Oslo, Oslo, Norway Anni Vedeler, Department of Biomedicine, University of Bergen, Bergen, Norway Jaakko Saraste, Department of Biomedicine, University of Bergen, Bergen, Norway Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Type 1 diabetes is an autoimmune disorder characterized by a lack of insulin production by the beta cells of the pancreas. This lack of insulin causes a variety of systemic effects on whole-body metabolism. Poorly managed type 1 diabetes can lead to cardiovascular disease, diabetic neuropathy, and diabetic retinopathy. Increasingly, even well-managed type 1 diabetic patients show damage to peripheral organs related to complications from the disease. The central role of insulin in energy homeostasis also renders it an important signaling factor in the reproductive tract. type 1 diabetes has now been demonstrated to cause defects in sperm and testes. The aim of this review is to present the known effects of insulin’s role in the function of the male reproductive tract. These effects might be mediated through hormonal alterations in the hypothalamic pituitary gonadal axis or through the direct interaction of insulin on the testes and sperm cells. Although fertility complications also occur in type 2 diabetic males, this review will focus on the defects specifically linked with the lack of insulin seen in type 1 diabetes. Content Type Journal Article Category Review Pages 1-9 DOI 10.1007/s00441-012-1387-7 Authors Erica L. Schoeller, Departments of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO 63110 USA Samantha Schon, Departments of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO 63110 USA Kelle H. Moley, Departments of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO 63110 USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Chronic Chagas’ disease is frequently characterized by massive myenteric neuron loss resulting in megacolon with severely and irreversibly disturbed motility. Here, we focused on two submucosal neuron populations, immunoreactive for calretinin (CALR) or somatostatin (SOM), and their respective mucosal nerve fibres in chagasic megacolon. Surgically removed megacolonic segments of seven chagasic patients were compared with seven age- and region-matched non-chagasic control segments. Evaluation included immunohistochemical triple-staining of cryosections for CALR, SOM and peripherin or for CALR and vasoactive intestinal peptide (VIP) and of submucosal whole-mounts for CALR, SOM and the pan-neuronal marker anti-HuC/D. Submucosal neuron counts in chagasic tissue revealed neuron numbers reduced to 51.2 % of control values. In cryosections, nerve fibre area measurements revealed 8.6 % nerve fibre per mucosal area in control segments, but this value decreased to 1.5 % in megacolonic segments. In both evaluations, a disproportionate decrease of SOM-reactive nerve elements was observed. The proportions of SOM-positive neurons related to the total neuron number declined to 2 % (control 10 %) and the proportion of SOM-reactive mucosal nerve fibres related to the whole mucosal area to 0.014 % (control 1.8 %)in chagasic tissue. The second set of cryosections revealed extensive colocalization of CALR with VIP in both surviving submucosal perikarya and mucosal nerve fibres. We suggest that VIP, a neuroprotective and neuroeffectory peptide typically contained in submucosal neurons, allows both the VIP-containing neurons to endure and the patients to survive by maintaining their mucosal barrier, despite the almost complete loss of colonic motility for decades. Content Type Journal Article Category Regular Article Pages 1-9 DOI 10.1007/s00441-012-1406-8 Authors Samir Jabari, Institute of Anatomy I, University of Erlangen-Nuremberg, Krankenhausstr. 9, 91054 Erlangen, Germany Alexandre B. M. da Silveira, Human Anatomy Sector, ICBIM, Universidade Federal de Uberlândia, Minas Gerais, Postal Code 38.400-902 Brazil Enio C. de Oliveira, Department of Surgery, Medical School, Universidade Federal de Goiás, Goiânia, Postal Code 74.605-020 Brazil Salustiano G. Neto, Department of Surgery, Medical School, Universidade Federal de Goiás, Goiânia, Postal Code 74.605-020 Brazil Karl Quint, Institute for Surgical Research, University of Marburg, Baldingerstr., 35043 Marburg, Germany Winfried Neuhuber, Institute of Anatomy I, University of Erlangen-Nuremberg, Krankenhausstr. 9, 91054 Erlangen, Germany Axel Brehmer, Institute of Anatomy I, University of Erlangen-Nuremberg, Krankenhausstr. 9, 91054 Erlangen, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Spermatozoa must translate information from their environment and the egg to achieve fertilization in sexually reproducing animals. These tasks require decoding a variety of signals in the form of intracellular Ca 2+ changes. As TRP channels constitute a large family of versatile multi-signal transducers, they are interesting subjects in which to explore their possible participation in sperm function. Here, we review the evidence for their presence and involvement in sperm motility, maturation, and the acrosome reaction, an exocytotic process required for sperm–egg fusion. Since store-operated Ca 2+ entry (SOCE) has been proposed to play an important role in these three functions, the main proteins responsible for this transport (STIM and ORAI) and their interaction with TRPs are also discussed. Improving our tools to solve infertility, improve animal breeding, and preserve biodiversity requires a better understanding of how Ca 2+ is regulated in spermatozoa. Content Type Journal Article Category Review Pages 1-16 DOI 10.1007/s00441-012-1397-5 Authors Alberto Darszon, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Claudia Sánchez-Cárdenas, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Gerardo Orta, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Ana A. Sánchez-Tusie, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Carmen Beltrán, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Ignacio López-González, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Gisela Granados-González, Facultad de Ciencias. Universidad Autónoma del Estado de México, Toluca Edo. de México, CP 50000 Cuernavaca, México Claudia L. Treviño, Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 62210 Cuernavaca, México Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Dentin sialoprotein (DSP) is a major non-collagenous protein in dentin. Mutation studies in human, along with gene knockout and transgenic experiments in mice, have confirmed the critical role of DSP for dentin formation. Our previous study reported that DSP is processed into fragments in mouse odontoblast-like cells. In order to gain insights into the function of DSP fragments, we further evaluated the expression pattern of DSP in the mouse odontoblast-like cells using immunohistochemistry and western blot assay with antibodies against the NH 2 -terminal and COOH-terminal regions of DSP. Then, the distribution profiles of the DSP NH 2 -terminal and COOH-terminal fragments and osteopontin (OPN) were investigated in mouse teeth at different ages by immunohistochemistry. In the odontoblast-like cells, multiple low molecular weight DSP fragments were detected, suggesting that part of the DSP protein was processed in the odontoblast-like cells. In mouse first lower molars, immunoreactions for anti-DSP-NH 2 antibody were intense in the predentin matrix but weak in mineralized dentin; in contrast, for anti-DSP-COOH antibody, strong immunoreactions were found in mineralized dentin, in particular dentinal tubules but weak in predentin. Therefore, DSP NH 2 -terminal and COOH-terminal fragments from odontoblasts were secreted to different parts of teeth, suggesting that they may play distinct roles in dentinogenesis. Meanwhile, both DSP antibodies showed weak staining in reactionary dentin (RD), whereas osteopontin (OPN) was clearly positive in RD. Therefore, DSP may be less crucial for RD formation than OPN. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1418-4 Authors Guohua Yuan, Key Laboratory for Oral Biomedicine of Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People’s Republic of China 430079 Guobin Yang, Key Laboratory for Oral Biomedicine of Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People’s Republic of China 430079 Guangtai Song, Key Laboratory for Oral Biomedicine of Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People’s Republic of China 430079 Zhi Chen, Key Laboratory for Oral Biomedicine of Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei, People’s Republic of China 430079 Shuo Chen, Department of Developmental Dentistry, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The robust axon regeneration that occurs following peripheral nerve injury is driven by transcriptional activation of the regeneration program and by the expression of a wide range of downstream effector molecules from neuropeptides and neurotrophic factors to adhesion molecules and cytoskeletal adaptor proteins. These regeneration-associated effector molecules regulate the actin-tubulin machinery of growth-cones, integrate intracellular signalling and stimulatory and inhibitory signals from the local environment and translate them into axon elongation. In addition to the neuronally derived molecules, an important transcriptional component is found in locally activated Schwann cells and macrophages, which release a number of cytokines, growth factors and neurotrophins that support neuronal survival and axonal regeneration and that might provide directional guidance cues towards appropriate peripheral targets. This review aims to provide a comprehensive up-to-date account of the transcriptional regulation and functional role of these effector molecules and of the information that they can give us with regard to the organisation of the regeneration program. Content Type Journal Article Category Review Pages 1-12 DOI 10.1007/s00441-012-1416-6 Authors Smriti Patodia, Centre for Perinatal Brain Protection and Repair, University College London, Chenies Mews 86-96, London, WC1E 6HX UK Gennadij Raivich, Centre for Perinatal Brain Protection and Repair, University College London, Chenies Mews 86-96, London, WC1E 6HX UK Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    VACM-1, a cul5 gene product, when overexpressed in vitro, has an antiproliferative effect. In vivo, VACM-1/cul5 is present in tissues involved in the regulation of water balance. Neither proteins targeted for VACM-1/cul5-specific degradation nor factors that may regulate its expression in those tissues have been studied. To identify genes that may be misregulated by VACM-1 cDNA, we performed microarray analysis. Our results indicate that in cos-1 cells transfected with VACM-1 cDNA, mRNA levels for several genes, including AQP1, were decreased when compared to the control group. Our results also indicate that in cos-1 cells transfected with VACM-1 cDNA, endogenous AQP1 protein was decreased about 6-fold when compared to the controls. To test the hypothesis that VACM-1/cul5 may be regulated by conditions that compromise water homeostasis in vivo, we determined if 24 h of water deprivation affects VACM-1/cul5 levels or the effect of VACM-1/cul5 on AQP1. VACM-1 mRNA and protein levels were significantly higher in rat mesenteric arteries, skeletal muscle and the heart ventricle but not in the heart atrium from 24-h water-deprived rats when compared to the controls. Interestingly, 24 h of water deprivation increased modification of VACM-1 by an ubiquitin-like protein, Nedd8, essential for cullin-dependent E3 ligase activity. Although water deprivation did not significantly change AQP1 levels in the mesenteric arteries, AQP1 protein concentrations were inversely correlated with the ratio of the VACM-1 to Nedd8-modified VACM-1. These results suggest that VACM-1/cul5 may regulate endothelial AQP1 concentration both in vivo and in vitro. Content Type Journal Article Category Regular Article Pages 1-13 DOI 10.1007/s00441-012-1419-3 Authors Alyssa E. Johnson, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Isabelle P. Le, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Bradley T. Andresen, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Joseph Stodola, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Gary L. Dewey, Holland Christian High School, Holland, MI 49422-9000, USA Shirley B. Dean, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA James Resau, Van Andel Research Institute, 333 Bostwick NE, Grand Rapids, MI 49503, USA Pete Haak, Van Andel Research Institute, 333 Bostwick NE, Grand Rapids, MI 49503, USA Travis Ruch, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Ashleigh Sartor, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Inara Lazdins, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Christopher C. Barney, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Maria A. Burnatowska-Hledin, Departments of Biology and Chemistry, Hope College, Holland, MI 49422-9000, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Inflammation is considered to be an important contributor to secondary damage after spinal cord injury (SCI). This secondary damage leads to further exacerbation of tissue loss and functional impairments. The immune responses that are triggered by injury are complex and are mediated by a variety of factors that have both detrimental and beneficial effects. In this review, we focus on the diverse effects of the phospholipase A 2 (PLA 2 ) superfamily and the downstream pathways that generate a large number of bioactive lipid mediators, some of which have pro-inflammatory and demyelinating effects, whereas others have anti-inflammatory and pro-resolution properties. For each of these lipid mediators, we provide an overview followed by a discussion of their expression and role in SCI. Where appropriate, we have compared the latter with their role in other neurological conditions. The PLA 2 pathway provides a number of targets for therapeutic intervention for the treatment of SCI and other neurological conditions. Content Type Journal Article Category Review Pages 1-19 DOI 10.1007/s00441-012-1430-8 Authors Samuel David, Center for Research in Neuroscience, The Research Institute of the McGill University Health Center, Livingston Hall, Room L7-210, 1650 Cedar Ave., Montreal, Quebec, Canada H3G 1A4 Andrew D. Greenhalgh, Center for Research in Neuroscience, The Research Institute of the McGill University Health Center, Livingston Hall, Room L7-210, 1650 Cedar Ave., Montreal, Quebec, Canada H3G 1A4 Rubèn López-Vales, Departament de Biologia Cellular, Fisiologia i Immunologia, Institut de Neurociències, CIBERNED, Universitat Autònoma de Barcelona, 08193 Bellaterra, Catalonia, Spain Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The blood-brain barrier (BBB) prevents free access of circulating molecules to the brain and maintains a specialized brain environment to protect the brain from blood-derived bioactive and toxic molecules; however, the circumventricular organs (CVOs) have fenestrated vasculature. The fenestrated vasculature in the sensory CVOs, including the organum vasculosum of lamina terminalis (OVLT), subfornical organ (SFO) and area postrema (AP), allows neurons and astrocytes to sense a variety of plasma molecules and convey their information into other brain regions and the vasculature in the secretory CVOs, including median eminence (ME) and neurohypophysis (NH), permits neuronal terminals to secrete many peptides into the blood stream. The present study showed that vascular permeability of low-molecular-mass tracers such as fluorescein isothiocyanate (FITC) and Evans Blue was higher in the secretory CVOs and kidney as compared with that in the sensory CVOs. On the other hand, vascular permeability of high-molecular-mass tracers such as FITC-labeled bovine serum albumin and Dextran 70,000 was lower in the CVOs as compared with that in the kidney. Prominent vascular permeability of low- and high-molecular-mass tracers was also observed in the arcuate nucleus. These data demonstrate that vascular permeability for low-molecular-mass molecules is higher in the secretory CVOs as compared with that in the sensory CVOs, possibly for large secretion of peptides to the blood stream. Moreover, vascular permeability for high-molecular-mass tracers in the CVOs is smaller than that of the kidney, indicating that the CVOs are not totally without a BBB. Content Type Journal Article Category Regular Article Pages 1-15 DOI 10.1007/s00441-012-1421-9 Authors Shoko Morita, Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, 606-8585 Japan Seiji Miyata, Department of Applied Biology, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, 606-8585 Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The purpose of this work was to determine if and where Angiopoietin-like genes are expressed in the mouse uterus during the implantation period of pregnancy and to determine if uterine expression of such genes is controlled by estrogen or progesterone. We found that all six known murine angiopoietin-like genes were expressed in the mouse uterus during implantation. The expression of four genes was controlled by either estrogen or progesterone. Only the levels of angiopoietin-like 4 ( Angptl4 ) mRNA dramatically increased in implantation segments of the uterus during decidualization and was conceptus-independent. Due to this increased expression and the fact that angiopoietin-like 4 protein plays a role in lipid metabolism and angiogenesis in other tissues, only the expression of Angptl4 was further examined in the uterus and developing placenta. Angptl4 mRNA was localized to subpopulations of the endometrial stromal fibroblast and endothelial cell populations during decidualization. It was also localized to the ectoplacental cone, trophoblast giant cells and parietal endoderm of the conceptus at this time. By mid-pregnancy, Angptl4 mRNA was localized mainly to the mesometrial lymphoid aggregate region plus mesometrial endothelial cells of the uterus, as well as in various cell types of the conceptus. Additional work showed that Angptl4 expression increases in mouse endometrial stromal cells as they undergo decidualization in vitro. As in other cell types, the expression of Angptl4 in endometrial stromal cells was increased in response to an agonist of the peroxisome proliferator activated receptors. Taken together, the results of this work support the hypothesis that locally expressed Angptl4 might play a role in local uterine/placental lipid metabolism and vascular changes during implantation and thus provide a basis for future research. Content Type Journal Article Category Regular Article Pages 1-13 DOI 10.1007/s00441-012-1337-4 Authors Charles A. Scott, Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA Doan van Huyen, Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA Brent M. Bany, Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois, USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The skin constitutes a formidable barrier against commensal and pathogenic bacteria, which permanently and transiently colonise the skin, respectively. Commensal and pathogenic species inhabiting skin both express proteases. Whereas proteases secreted by commensals contribute to homeostatic bacterial coexistence on skin, proteases from pathogenic bacteria are used as virulence factors, helping them colonise skin with breached integrity of the epithelial layer. From these initial sites of colonisation, pathogens can disseminate into deeper layers of skin, possibly leading to the spread of infection. Secreted bacterial proteases probably play an important role in this process and in the deterrence of innate defence mechanisms. For example, Staphylococcus aureus proteases are essential for changing the bacterial phenotype from adhesive to invasive by degrading adhesins on the bacterial cell surface. Secreted staphylococcal proteases mediate pathogen penetration by degrading collagen and elastin, essential components of connective tissue in the dermis. The activation of the contact system and kinin generation by Streptococcus pyogenes and S. aureus proteases contributes to an inflammatory reaction manifested by oedema, redness and pain. Kinin-enhanced vascular leakage might help bacteria escape into the circulation thereby causing possible systemic dissemination of the infection. The inflammatory reaction can also be fueled by the activation of protease-activated receptors on keratinocytes. Concomitantly, bacterial proteases are involved in degrading antimicrobial peptides, disarming the complement system and neutrophils and preventing the infiltration of the infected sites with immune cells by inactivation of chemoattractants. Together, this provides protection for colonising and/or invading pathogens from attack by antibacterial forces of the skin. Content Type Journal Article Category Review Pages 1-13 DOI 10.1007/s00441-012-1355-2 Authors Joanna Koziel, Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387 Kraków, Poland Jan Potempa, Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30-387 Kraków, Poland Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide with highly potent neurotrophic and neuroprotective effects. PACAP and its receptors occur in the retina and PACAP has been applied in animal models of metabolic retinal disorders to reduce structural and functional damage. Furthermore, PACAP has been implicated as a potential anti-diabetic peptide. Our aim has been to investigate, by using a complex morphological, immunochemical and molecular biological approach, whether PACAP attenuates diabetic retinopathy. Diabetes was induced in rats with a single streptozotocin injection. PACAP was injected intravitreally into one eye (100 pmol) three times during the last week of a 3-week survival period. Retinas were processed for the following procedures: routine histology, immunohistochemistry (single and double labeling, whole-mount), quantitative reverse transcription with the polymerase chain reaction and Western blotting. Cone photoreceptors and dopaminergic amacrine and ganglion cells degenerated in diabetic retinas and glial fibrillary acidic protein were upregulated in Müller glial cells. The number of cones, the length of their outer segments and the cell number in the ganglion cell layer were decreased. PACAP ameliorated these structural changes. Moreover, PACAP increased the levels of PAC1-receptor and tyrosine-hydroxylase as detected by molecular biological methods. Thus, PACAP has significant protective effects in the diabetic retina. PACAP treatment attenuates neuronal cell loss in diabetic retinopathy, the protective effects of PACAP probably being mediated through the activation of PAC1-receptor. These results suggest that PACAP has a therapeutic potential in diabetic retinopathy. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1349-0 Authors Krisztina Szabadfi, Department of Experimental Zoology and Neurobiology, Institute of Biology, University of Pecs, Ifjusag Street 6, 7624 Pecs, Hungary Tamas Atlasz, Department of Sportbiology, University of Pecs, Pecs, Hungary Peter Kiss, Department of Anatomy, PTE-MTA PACAP Research Team, University of Pecs, Pecs, Hungary Dora Reglodi, Department of Anatomy, PTE-MTA PACAP Research Team, University of Pecs, Pecs, Hungary Aliz Szabo, Department of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary Krisztina Kovacs, Department of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary Balint Szalontai, Department of Plant Physiology, University of Pecs, Pecs, Hungary Gyorgy Setalo, Department of Medical Biology, University of Pecs, Pecs, Hungary Eszter Banki, Department of Anatomy, PTE-MTA PACAP Research Team, University of Pecs, Pecs, Hungary Katalin Csanaky, Department of Anatomy, PTE-MTA PACAP Research Team, University of Pecs, Pecs, Hungary Andrea Tamas, Department of Anatomy, PTE-MTA PACAP Research Team, University of Pecs, Pecs, Hungary Robert Gabriel, Department of Experimental Zoology and Neurobiology, Institute of Biology, University of Pecs, Ifjusag Street 6, 7624 Pecs, Hungary Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Carvajal syndrome is a rare syndrome with woolly hair, palmoplantar keratosis and dilated cardiomyopathy. The inheritance of the mutation is autosomal recessive. As a causal gene, the desmoplakin gene ( DSP ) has so far been identified; it encodes an essential component of desmosomes, a cell-cell structure aimed at keeping cells attached to each other in tissues in which cells are often exposed to strong shear forces. Recently, familial cases of an autosomal dominant Carvajal syndrome were documented with a new feature: hypo/oligodontia. A mutation in the DSP gene was also evidenced in these latter cases. A patient was seen for cardiogenetic consultation at the University Hospital of Lyon with cardiac failure involving first degree atrioventricular block, complete left bundle branch block, non-compaction of the apex of the left ventricle and a dilated cardiomyopathy. A coronarography disclosed a complete thrombosis of the right coronary artery. At examination, he had also woolly hair, mild palmoplantar keratosis and missing teeth (essentially molars and premolars). His family history was uninformative. His DNA was screened for mutations in the DSP and plakoglobin genes but no mutation could be found. This case suggests that Carvajal syndrome with hypo/oligodontia is a heterogeneous condition in which genes other than DSP might be involved, although we cannot rule out a mutation in this gene consisting in a deletion of a single exon or a gene rearrangement. Content Type Journal Article Category Regular Article Pages 1-4 DOI 10.1007/s00441-012-1351-6 Authors Nancy Nehme, Laboratoire Cardiogénétique, Centre de Biologie et Pathologie Est, Groupe Hospitalier Est, 59 Boulevard Pinel, 69677 Bron, France Rajae El Malti, Laboratoire Cardiogénétique, Centre de Biologie et Pathologie Est, Groupe Hospitalier Est, 59 Boulevard Pinel, 69677 Bron, France Nathalie Roux-Buisson, Laboratoire de Biochimie et Biologie Moléculaire, Centre Hospitalier Universitaire de Grenoble, BP 217, 38043 La Tronche, France Jean-Raymond Caignault, Service de Cardiologie et Pathologie Vasculaire, Hôpital d’Instruction des Armées Desgenettes, 108 Boulevard Pinel, 69275 Lyon cedex 03, France Patrice Bouvagnet, Laboratoire Cardiogénétique, Centre de Biologie et Pathologie Est, Groupe Hospitalier Est, 59 Boulevard Pinel, 69677 Bron, France Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Zona pellucida (ZP) is a glycoproteinaceous translucent matrix that surrounds the mammalian oocyte and plays a critical role in the accomplishment of fertilization. In humans, it is composed of 4 glycoproteins designated as ZP1, ZP2, ZP3 and ZP4, whereas mouse ZP is composed of ZP1, ZP2 and ZP3 ( Zp4 being a pseudogene). In addition to a variable sequence identity of a given zona protein among various species, human ZP1 and ZP4 are paralogs and mature polypeptide chains share an identity of 47%. Employing either affinity purified native or recombinant human zona proteins, it has been demonstrated that ZP1, ZP3 and ZP4 bind to the capacitated human spermatozoa and induce an acrosome reaction, whereas in mice, ZP3 acts as the putative primary sperm receptor. Human ZP2 only binds to acrosome-reacted spermatozoa and thus may be acting as a secondary sperm receptor. In contrast to O -linked glycans of ZP3 in mice, N -linked glycans of human ZP3 and ZP4 are more relevant for induction of the acrosome reaction. Recent studies suggest that Sialyl-Lewis x sequence present on both N - and O -glycans of human ZP play an important role in human sperm–egg binding. There are subtle differences in the downstream signaling events associated with ZP3 versus ZP1/ZP4-mediated induction of the acrosome reaction. For example, ZP3 but not ZP1/ZP4-mediated induction of the acrosome reaction is dependent on the activation of the G i protein-coupled receptor. Thus, various studies suggest that, in contrast to mice, in humans more than one zona protein binds to spermatozoa and induces an acrosome reaction. Content Type Journal Article Category Review Pages 1-14 DOI 10.1007/s00441-011-1319-y Authors Satish K. Gupta, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Beena Bhandari, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Abhinav Shrestha, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Bichitra K. Biswal, Protein Crystallography Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067 India Chetna Palaniappan, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Sudha Saryu Malhotra, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Neha Gupta, Reproductive Cell Biology Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110067 India Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Regenerative therapies, including cell injection and bioengineered tissue transplantation, have the potential to treat severe heart failure. Direct implantation of isolated skeletal myoblasts and bone-marrow-derived cells has already been clinically performed and research on fabricating three-dimensional (3-D) cardiac grafts using tissue engineering technologies has also now been initiated. In contrast to conventional scaffold-based methods, we have proposed cell sheet-based tissue engineering, which involves stacking confluently cultured cell sheets to construct 3-D cell-dense tissues. Upon layering, individual cardiac cell sheets integrate to form a single, continuous, cell-dense tissue that resembles native cardiac tissue. The transplantation of layered cardiac cell sheets is able to repair damaged hearts. As the next step, we have attempted to promote neovascularization within bioengineered myocardial tissues to overcome the longstanding limitations of engineered tissue thickness. Finally, as a possible advanced therapy, we are now trying to fabricate functional myocardial tubes that may have a potential for circulatory support. Cell sheet-based tissue engineering technologies therefore show an enormous promise as a novel approach in the field of myocardial tissue engineering. Content Type Journal Article Category Review Pages 1-8 DOI 10.1007/s00441-011-1267-6 Authors Hidekazu Sekine, Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan Tatsuya Shimizu, Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan Teruo Okano, Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The central nervous system (CNS) has been traditionally considered as an organ that fails to regenerate in response to injury. Indeed, the lesioned CNS faces a number of obstacles during regeneration, including an overall non-permissive environment for axonal regeneration. However, research during the last few decades has identified axon sprouting as an anatomical correlate for the regenerative capability of the CNS to establish new connections. The immunoglobulin superfamily member L1CAM has been shown to promote the capability of neurons for regenerative axon sprouting and to improve behavioral outcomes after CNS injury. Here, we discuss the cell-autonomous role of L1CAM for axon sprouting in experimental rodent injury models and highlight the molecular interactions of L1CAM with ankyrins, ezrin-radixin-moesin proteins and the Sema3A/Neuropilin ligand-receptor complex in the context of axonal branching. Content Type Journal Article Category Review Pages 1-10 DOI 10.1007/s00441-012-1345-4 Authors Michael K. E. Schäfer, Klinik für Anästhesiologie, Universitätsmedizin Mainz, 55131 Mainz, Germany Michael Frotscher, Zentrum für Molekulare Neurobiologie Hamburg, 20246 Hamburg, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Apart from the well-known biologically active angiotensin II, other biologically active angiotensins have been discovered, including angiotensin IV and angiotensin-(1–7). Some years ago, we and others discovered that the Mas proto-oncogene encodes a receptor that is essential for angiotensin-(1–7) signaling. Angiotensin-(1–7) is not only expressed in the periphery but also within the brain. Based on that, we examined the distribution of Mas within the murine brain, using an antibody directed against the 3 rd cytoplasmic loop of the receptor protein. Strongest Mas protein expression was detected in the dentate gyrus of the hippocampus and within the piriform cortex. However, Mas protein expression is not restricted to these areas, since Mas immunopositive neurons were also seen in different parts of the cortex, hippocampus, amygdala, basal ganglia, thalamus and hypothalamus. Based on the expression of Mas protein in the cortex and the limbic system, angiotensin-(1–7) signaling may play a role in synaptic plasticity, learning, memory and emotion, as has been described for angiotensin II and IV. Content Type Journal Article Category Regular Article Pages 1-7 DOI 10.1007/s00441-012-1354-3 Authors M. Freund, Institute of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Ernst Moritz Arndt University of Greifswald, Friedrich-Löffler-Straße-23c, 17487 Greifswald, Germany T. Walther, Centre Cardiovascular and Metabolic Research (CCMR), Hull York Medical School, University of Hull, Hull, HU6 7RX UK O. von Bohlen und Halbach, Institute of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Ernst Moritz Arndt University of Greifswald, Friedrich-Löffler-Straße-23c, 17487 Greifswald, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Visual impairment severely affects the quality of life of patients and their families and is also associated with a deep economic impact. The most common pathologies responsible for visual impairment and legally defined blindness in developed countries include age-related macular degeneration, glaucoma and diabetic retinopathy. These conditions share common pathophysiological features: dysfunction and loss of retinal neurons. To date, two main approaches are being taken to develop putative therapeutic strategies: neuroprotection and cell replacement. Cell replacement is a novel therapeutic approach to restore visual capabilities to the degenerated adult neural retina and represents an emerging field of regenerative neurotherapy. The discovery of a population of proliferative cells in the mammalian retina has raised the possibility of harnessing endogenous retinal stem cells to elicit retinal repair. Furthermore, the development of suitable protocols for the reprogramming of differentiated somatic cells to a pluripotent state further increases the therapeutic potential of stem-cell-based technologies for the treatment of major retinal diseases. Stem-cell transplantation in animal models has been most effectively used for the replacement of photoreceptors, although this therapeutic approach is also being used for inner retinal pathologies. In this review, we discuss recent advances in the development of cell-replacement approaches for the treatment of currently incurable degenerative retinal diseases. Content Type Journal Article Category Review Pages 1-12 DOI 10.1007/s00441-012-1335-6 Authors Christian W. Schmeer, Hans Berger Clinic of Neurology, University Hospital Jena, Erlanger Allee 101, 07747 Jena, Germany Stefanie G. Wohl, Hans Berger Clinic of Neurology, University Hospital Jena, Erlanger Allee 101, 07747 Jena, Germany Stefan Isenmann, Department of Neurology, HELIOS Klinikum, Wuppertal, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description: Endogenous musculoskeletal tissue regeneration Content Type Journal Article Category Editorial Pages 1-4 DOI 10.1007/s00441-012-1357-0 Authors Dietmar W. Hutmacher, Institute of Health and Biomedical Innovation, Queensland University of Technology & George W. Woodruff School of Mechanical Engineering Georgia Institute of Technology, Atlanta, Ga., USA Georg Duda, Julius Wolff Institute and Center for Musculoskeletal Surgery & Berlin-Brandenburg Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin, Berlin, Germany Robert E. Guldberg, Parker H. Petit Institute for Bioengineering and Bioscience & George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, Ga., USA Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    The musculoskeletal system is a tight network of many tissues. Coordinated interplay at a biochemical level between tissues is essential for development and repair. Traumatic injury usually affects several tissues and represents a large challenge in clinical settings. The current demand for potent growth factors in such applications thus accompanies the keen interest in molecular mechanisms and orchestration of tissue formation. Of special interest are multitasking growth factors that act as signals in a variety of cell types, both in a paracrine and in an autocrine manner, thereby inducing cell differentiation and coordinating not only tissue assembly at specific sites but also maturation and homeostasis. We concentrate here on bone morphogenetic proteins (BMPs), which are important crosstalk mediators known for their irreplaceable roles in vertebrate development. The molecular crosstalk during embryonic musculoskeletal tissue formation is recapitulated in adult repair. BMPs act at different levels from the initiation to maturation of newly formed tissue. Interestingly, this is influenced by the spatiotemporal expression of different BMPs, their receptors and co-factors at the site of repair. Thus, the regenerative potential of BMPs needs to be evaluated in the context of highly connected tissues such as muscle and bone and might indeed be different in more poorly connected tissues such as cartilage. This highlights the need for an understanding of BMP signaling across tissues in order to eventually improve BMP regenerative potential in clinical applications. In this review, the distinct members of the BMP family and their individual contribution to musculoskeletal tissue repair are summarized by focusing on their paracrine and autocrine functions. Content Type Journal Article Category Review Pages 1-24 DOI 10.1007/s00441-011-1283-6 Authors Karen Ruschke, Institute for Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Christian Hiepen, Institute for Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Jessica Becker, Institute for Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Petra Knaus, Institute for Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X

Description:    Diabetic dyslipidemia is characterized by increased circulatory very-low-density lipoprotein (VLDL) levels. Aldosterone, apart from its role in fluid and electrolyte homeostasis, has also been implicated in insulin resistance and myocardial fibrosis. The impact of VLDL as a potential risk factor for aldosterone-mediated cardiovascular injury in diabetes mellitus, however, remains to be investigated. We have therefore studied native and modified VLDL-mediated steroidogenesis and its underlying molecular mechanisms in human adrenocortical carcinoma cells, NCI H295R. Native VLDL (natVLDL), isolated from healthy volunteers, was subjected to in vitro modification with glucose (200 mmol/l) or sodium hypochlorite (1.5 mmol/l) for preparation of glycoxidized and oxidized VLDL, respectively. VLDL treatment induced steroidogenesis in both a concentration- and time-dependent manner. Native and glycoxidized VLDL (50 μg/ml) were almost two-fold more potent in adrenocortical aldosterone release than angiotensin II (100 nmol/l). These forms of VLDL significantly augmented transcriptional regulation of aldosterone synthase ( Cyp11B2 ), partially through scavenger receptor class B type I, as evident from the effect of BLT-1. In contrast to glycoxidized VLDL, oxidized VLDL significantly attenuated the stimulatory effect of natVLDL on adrenocortical hormone synthesis. Moreover, treatment with specific pharmacological inhibitors (H89, U0126, AG490) provided supporting evidence that VLDL, irrespective of modification, presumably recruited PKA, ERK1/2 and Jak-2 for steroid hormone release through modulation of Cyp11B2 mRNA level. In conclusion, this study demonstrates a novel insight into intracellular mechanism of VLDL-mediated aldosterone synthesis through transcriptional regulation of steroidogenic acute regulatory protein ( StAR ) and Cyp11B2 expression in human adrenocortical carcinoma cell line. Content Type Journal Article Category Regular Article Pages 1-10 DOI 10.1007/s00441-012-1346-3 Authors Sarama Saha, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Stefan R. Bornstein, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Juergen Graessler, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Steffi Kopprasch, Department of Internal Medicine III, Carl Gustav Carus Medical School, Technical University of Dresden, Fetscherstraße 74, 01307 Dresden, Germany Journal Cell and Tissue Research Online ISSN 1432-0878 Print ISSN 0302-766X