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  • 1
    Electronic Resource
    Electronic Resource
    PML-containing nuclear bodies: Their spatial distribution in relation to other nuclear components (1996)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 63 (1996), S. 280-291 
    Details
    ISSN: 0730-2312
    Keywords: nuclear bodies ; PML ; confocal microscopy ; image restoration ; RNA ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The PML protein is a human growth suppressor concentrated in 10 to 20 nuclear bodies per nucleus (PML bodies). Disruption of the PML gene has been shown to be related to acute promyelocytic leukaemia (APL). To obtain information about the function of PML bodies we have investigated the 3D-distribution of PML bodies in the nucleus of T24 cells and compared it with the spatial distribution of a variety of other nuclear components, using fluorescence dual-labeling immunocytochemistry and confocal microscopy. Results show that PML bodies are not enriched in nascent RNA, the splicing component U2-snRNP, or transcription factors (glucocorticoid receptor, TFIIH, and E2F). These results show that PML bodies are not prominent sites of RNA synthesis or RNA splicing. We found that a large fraction of PML bodies (50 to 80%) is closely associated with DNA replication domains during exclusively middle-late S-phase. Furthermore, in most cells that we analysed we found at least one PML body was tightly associated with a coiled body. In the APL cell line NB4, the PML gene is fused with the RARα gene due to a chromosomal rearrangement. PML bodies have disappeared and the PML antigen, i.e., PML and the PML-RAR fusion protein, is dispersed in a punctated pattern throughout the nucleoplasm. We showed that in NB4 cells the sites that are rich in PML antigen significantly colocalize with sites at which nascent RNA accumulates. This suggests that, in contrast to non-APL cells, in NB4 cells the PML antigen is associated with sites of transcription. The implications of these findings for the function of PML bodies are consistent with the idea that PML bodies are associated with specific genomic loci. © 1996 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    Transforming growth factor-β in the early mouse embryo: Implications for the regulation of muscle formation and implantation (1993)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 14 (1993), S. 212-224 
    Details
    ISSN: 0192-253X
    Keywords: Embryonic stem cells ; differentiation ; organogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a search for functions of transforming growth factor-β during early embryonic development we used two different experimental approaches. In the first we made use of embryonic stem (ES) cells. ES cells in culture differentiate to derivatives of all three germ layers and mimic some aspects of organogenesis when grown as aggregates in suspension to form embryoid bodies. Differentiation procedes further when the embryold bodies attach to suitable substrates. Muscle and neuronal cells are among the most readily identified cell types then formed. We examined the effect of all-trans retinoic acid (RA) and members of the transforming growth factor-β family(TGF-βl, TGF-β2) under these conditions in an assay where single aggregates formed in hanging microdrops in medium supplemented with serum depleted of lipophilic substances which would include retinoids. Endoderm-like cells formed under all conditions tested. RA at concentrations of 108 M and 107 M induced the formation of neurons but in the absence of RA or at concentrations up to 10-9 M, neurons were not observed. Instead, beating muscle formed in about one-third of the plated aggregates; this was greatly reduced when RA concentrations increased above 10-9 M. Immunofluorescent staining for muscle specific myosin showed that two muscle cell types could be distinguished: elongated, non-contractile myoblasts and mononucleate flat cells. The mononucleate flat cells appeared to correspond with rhythmically contracting muscle. The number of non-contractile myoblasts increased 3-fold over controls in the presence of 10-9 M RA. TGF-βs increased the number of contractile and non-contractile muscle cells by a factor 3 to 7 over controls, depending on the TGF-β isoform added and the muscle cell type formed. TGF-β2 also invariably increased the rate at which contracting muscle cells were first observed in replated aggregates. The stimulatory effect of TGF-βs on the formation of mononucleate flat cells was completely abrogated by RA at 10-9 M while the number of myoblasts under similar conditions was unchanged. These data suggest that a complex interplay between retinoids and TGF-β isoforms may be involved in regulation of differentiation in early myogenesis.In the second approach, neutralizing polyclonal rabbit antibodies specific for TGF-β2 were injected into the cavity of mouse blastocysts 3.5 days post coitum (pc). After 1 day in culture, embryos were transferred to pseudopregnant females. The number of decidua, embryos and resorptions were counted at day 8.5-9.5 pc. Control antibody injected embryos implanted with high efficiency (87%) compared with anti-TGF-β2 injected embryos which implanted with an efficiency of only 43%. If empty decidua (resorptions) were included, the overall recovery was 71% and 32% for control and experimental embryos, respectively. Embryos that were recovered showed no overt macroscopic abnormalities. These results together impiy functions for TGF-βs in implantation as well as in later development of the embryo. © 1993Wiley-Liss, Inc.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020140308
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  • 3
    Electronic Resource
    Electronic Resource
    Nuclear transfer and electrofusion in bovine in vitro-matured/in vitro-fertilized embryos: Effect of media and electrical fusion parameters (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 36 (1993), S. 307-312 
    Details
    ISSN: 1040-452X
    Keywords: Micromanipulation ; IVM oocytes ; IVM/IVF donor embryos ; Cytochalasin B ; Nocodazole ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this study, micromanipulation and electrofusion conditions for the cloning of in vitro-produced bovine embryos (here after termed IVM/IVF embryos) derived from in vitro-matured (IVM) and in vitro-fertilized (IVF) oocytes were established. The effect of DC field strength on the fusion rate was tested in a model system using pronuclear stage embryos in which a cytoplasmic vesicle was removed and reinserted. Efficient fusion (80%) was obtained by applying a pulse of 1.75 kV/cm for 40 μsec. In vitro development of manipulated pronuclear stage embryos was as efficient as that of unmanipulated control embryos. Different fusion media were compared in the cloning procedure, using IVM oocytes as recipients and blastomeres from day 6 IVM/IVF donor embryos. Zimmermann cell fusion medium reduced the lysis of nuclear transfer embryos compared to F300 (5% vs. 25%). The effects of drugs disrupting the microfilaments and microtubuli were determined. Neither the addition of cytochalasin B (CCB) for 1 hr in the postfusion medium nor incubation of donor blastomeres with nocodazole had a significant effect on the fusion or cleavage rate of the nuclear transfer embryos. Additional experiments demonstrated that there was no difference in developmental potential between nuclear transfer embryos allowed to develop in vitro or in vivo and that the embryos gave a 15% pregnancy rate in recipient cattle. © 1993 Wiley-Liss, Inc.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080360304
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  • 4
    Electronic Resource
    Electronic Resource
    Distribution of the intermediate filament proteins vimentin, keratin, and desmin in the bovine ovary (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 41 (1995), S. 459-467 
    Details
    ISSN: 1040-452X
    Keywords: Vimentin ; Keratin ; Desmin ; Immunohis-tochemistry ; Ovary ; Bovine ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The distribution of the intermediate filament (IF) proteins desmin, keratin, and vimentin was studied immunohistochemically in bovine ovaries. Special attention was paid to granulosa cells to examine possible marked changes of IF distribution in relation to folliculogenesis during ovarian development. Therefore, ovaries were used from fetuses from 3 months of gestation onward, calves, heifers, and cows. In all ovaries, desmin immunoreactivity was restricted to smooth muscle cells in blood vessel walls. Keratin appeared a characteristic of the ovarian surface epithelium. Co-localization of keratin and vimentin was observed in the epithelium of rete ovarii tubules in fetuses and calves, and in cortical cord epithelium and pregranulosa cells of primordial follicles in fetuses at 3-7 months of gestation. Vimentin was demonstrated in endothelium and in fibroblasts. In addition, vimentin immunoreactivity was present in granulosa cells of primary, secondary, and antral follicles. In antral follicles, these granulosa cells mainly had an elongated appearance and either contained an oblong or a round nucleus. Those with an oblong nucleus were characteristic for atretic antral follicles. In nonatretic follicles, numerous vimentin immunore-active, elongated granulosa cells with a round nucleus were observed, especially in the peripheral granulosa layer and in small (〈3 mm in diameter) antral follicles. Additionally, in antral follicles, protrusions of vimentin-positive corona radiata cells were observed, that penetrated the zona pellucida to contact the oocyte. The data show that the distribution of vimentin containing IFs is associated with various aspects of granulosa cell activity, as mitosis, atresia, and intercellular transport. © 1995 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080410408
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  • 5
    Electronic Resource
    Electronic Resource
    Ultrastructural evidence for the axonal localization of caudodorsal cell hormone mRNA in the central nervous system of the mollusc Lymnaea stagnalis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 12-18 
    Details
    ISSN: 1059-910X
    Keywords: In situ hybridization ; Digoxigenin ; Electron microscopy ; Cryosections ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The technique of in situ hybridization has been used to evaluate the expression of an ovulation hormone mRNA (caudodorsal cell hormone; CDCH) in the central nervous system (CNS) of the mollusc Lymnaea stagnalis. Hybridization with radioactive as well as with nonradioactive labeled oligonucleotide and plasmid probes revealed a specific labeling on cell bodies of caudodorsal cells (CDCs), which are known to produce CDCH, on the light microscopical level. In addition, specific labeling was observed outside the cell bodies, as far as the cerebral commissure, where CDCH is released in the haemolymph. To investigate whether these signals represent an axonal localization of the CDCH mRNA, we performed in situ hybridization at the electron microscopical (EM) level. The results showed an intraaxonal localization of CDCH mRNA with digoxigenin labeled oligonucleotide and plasmid probes. Gold labeling was observed in secretion granules, and double labeling experiments showed that these granules also contain CDCH. This specific intragranular localization suggest that CDCH mRNA is transported through the axon and released by exocytosis in the haemolymph. © 1993 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070250104
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  • 6
    Electronic Resource
    Electronic Resource
    Immunocytochemical study of crustacean hyperglycemic hormone in the eyestalks of the prawn Palaemon serratus (Pennant) and some other Palaemonidae, in relation to variations in the blood glucose level (1984)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 182 (1984), S. 85-94 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: With the use of rabbit antisera against crustacean hyperglycemic hormone (CHH), it is possible to describe a distinct immunopositive reaction in a group of neurosecretory cells in the medulla terminalis ganglionic X-organ2 (MTGX2), in the MTGX-sinus gland tract, and in a considerable part of the sinus gland from several species of prawns belonging to the Palaemonidae. By introductory studies on the CHH system in Palaemon serratus, we can postulate a sequence in the activity cycle of the CHH-producing cells on the basis of differences in staining intensity of the immunoreaction and such morphometric parameters as cellular and nuclear diameter. By studying the CHH-producing system in combination with variations in the glucose level of the blood, an “inverse relationship” is observed between the number of immunoreactive cells and the blood glucose level during different periods of the year as well as during different stages of the molting cycle. A “shift in phase” of this correlation during the diurnal cycle suggests that several rhythmical phenomena may play a role in the regulation of glycemia in Crustacea.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1051820106
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  • 7
    Electronic Resource
    Electronic Resource
    Comparative microanatomy of the branchial sieve in three sympatric cyprinid species, related to filter-feeding mechanisms (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 73-87 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: According to the reducible-channel model of filter-feeding (Hoogenboezem et al., '91), small food particles are retained in the channels between the medial gill rakers, while the mesh size can be reduced by lowering the lateral gill rakers into these channels. This movement requires that all lateral gill rakers have a m. abductor branchiospinalis (MAB). MAB runs from the radii branchiales to the raker feet. It is present on the lateral side of all four gill arches of the cyprinids Abramis brama and Cyprinus carpio but only on the first arch of Blicca bjoerkna, Rutilus rutilus, Ctenopharyngodon idella, Aspius aspius, and Scardinius erythrophthalmus. Therefore, the latter species do not fulfill the structural requirement for the reducible-channel model, whereas A. brama and C. carpio do. Laboratory and field data confirm that A. brama and C. carpio can reduce their mesh size according to this model and are the better filter-feeders. The seven cyprinid species studied show the same principal microanatomy of their branchial sieve. M. abductor filamenti is a sheet of muscle fibers between the lateral radii branchiales and the ceratobranchial bone. M. branchialis superficialis is a specialized region of the subepithelial muscle fiber network, with origins along both sides of the ceratobranchial bone. The lateral gill rakers of the first gill arch differ conspicuously from all other rakers. They are longer and flattened, and they are tilted anteriorly. They probably form a sieve across the wide slit between the first gill arch and the operculum. The most revealing anatomical feature is the presence of MABs on gill arches 1-4. It might be a suitable bio-assay for identifying the better facultative filter-feeders among cyprinids. © 1994 Wiley-Liss, Inc.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052190109
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  • 8
    Electronic Resource
    Electronic Resource
    The innervation of some proboscis structures involved in feeding behavior of the blowfly (Calliphora vicina) (1985)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 186 (1985), S. 279-287 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The microtopology of the motoneurons involved in protraction and retraction of the proboscis of the blowfly (Calliphora vicina) has been studied. In addition, taste input from the labellar hairs was investigated. As a result of this study it appears that protraction movements are controlled by two while retraction movements are guided by three motoneurons on each side. The neurons in each group apear to be in ipsicontralateral communication with each other. The musculi protractores fulcri (MPF) probably contain a proprioceptive cell group which projects to the MPF motoneurons. It is proposed that the proboscis motor system can be modulated by proprioception as well as by chemosensory labellar input. Neurosecretory cells may be involved in adjusting muscle power.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1051860305
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  • 9
    Electronic Resource
    Electronic Resource
    Protein-DNA interactions at the H4-Site III upstream transcriptional element of a cell cycle regulated histone H4 gene: Differences in normal versus tumor cells (1992)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 49 (1992), S. 93-110 
    Details
    ISSN: 0730-2312
    Keywords: DNA-binding proteins ; Differentiation ; Distal promoter elements ; Proliferation ; Cell growth ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Upstream sequences of the H4 histone gene FO108 located between nt -418 to -213 are stimulatory for in vivo transcription. This domain contains one protein/DNA interaction site (H4-Site III) that binds factor H4UA-1. Based on methylation interference, copper-phenanthroline protection, and competition assays, we show that H4UA-1 interacts with sequences between nt -345 to -332 containing an element displaying sequence-similarity with the thyroid hormone response element (TRE). Using gel retardation assays, we also demonstrate that H4UA-1 binding activity is abolished at low concentrations of Zn2+ (0.75 mM), a characteristic shared with the thyroid hormone (TH) receptor DNA binding protein. Interestingly, phosphatase-treatment of nuclear proteins inhibits formation of the H4UA-1 protein/DNA complex, although a complex with higher mobility (H4UA-1b) can be detected; both complexes share identical protein-DNA contacts and competition behaviors. These findings suggest that phosphorylation may be involved in the regulation of H4-Site III protein/DNA interactions by directly altering protein/protein associations. H4-Site III interactions were examined in several cell culture systems during cell growth and differentiation. We find that H4UA-1 binding activity is present during the cell cycle of both normal diploid and transformed cells. However, during differentiation of normal diploid rat calvarial osteoblasts, we observe a selective loss of the H4UA-1/H4-Site III interaction, concomitant with an increase of the H4UA-1b/H4-Site III complex, indicating modifications in the heteromeric nature of protein/DNA interactions during downregulation of transcription at the cessation of proliferation. Transformed cells have elevated levels of H4UA-1, whereas H4UA-1b is predominantly present in normal diploid cells; this alteration in the ratio of H4UA-1 and H4UA-1b binding activities may reflect deregulation of H4-Site III interactions in transformed cells. We propose that H4-Site III interactions may contribute, together with protein/DNA interactions at proximal regulatory sequences, in determining the level of H4-FO108 histone gene transcription.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240490115
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  • 10
    Electronic Resource
    Electronic Resource
    Phenotypic transformation of normal rat kidney cells in a growth-factor-defined medium: Induction by a neuroblastoma-derived transforming growth factor independently of the EGF receptor (1985)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 123 (1985), S. 151-160 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Polypeptide growth factor activity in serum can be destroyed by treatment with dithiothreitol. When such growth-factor-inactivated serum is used as a supplement of culture media instead of regular serum, normal rat kidney (NRK) cells become quiescent unless defined polypeptide growth factors like insulin and epidermal growth factor (EGF) are added. On this basis a growth-factor-defined medium has been developed for NRK cells, which permits cell proliferation as rapidly as in media supplemented with serum, even at low cell densities. Moreover, cells can be serially passaged in this medium. NRK cells can be induced to grow in semisolid media when incubated with transforming growth factors. The growth-factor-defined medium permits soft agar growth experiments of NRK cells, without interference from polypeptide growth factors in serum. Using this assay system we have shown that EGF alone is unable to induce any degree of anchorage-independent growth in NRK cells. However, a recently identified transforming growth factor from mouse neuroblastoma cells which does not compete with EGF for receptor binding is able to induce progressively growing colonies of NRK cells in soft agar, even without additional EGF.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041230202
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