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  • Immunocytochemistry  (428)
  • Springer  (428)
  • American Meteorological Society
  • 1995-1999  (166)
  • 1990-1994  (262)
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  • 1
    Electronic Resource
    Electronic Resource
    Seasonal variations in the production of the egg-jelly macromolecule, a fucose sulphate glycoconjugate, by the accessory cells in the ovary of the sea urchin Hemicentrotus pulcherrimus (1994)
    Springer
    Development genes and evolution 203 (1994), S. 402-410 
    Details
    ISSN: 1432-041X
    Keywords: Sea urchin ; Egg jelly ; Ovary ; Development ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the sea urchin Hemicentrotus pulcherrimus, the egg-jelly macromolecule, a fucose sulphate glycoconjugate (FSG) that induces the acrosome reaction in spermatozoa, originates from the accessory cells in the ovary. In the present study we examined the seasonal variations in the distribution of FSG in the ovary by immunocytochemistry with a polyclonal antibody. An enzyme-linked immunosorbent assay indicated that FSG was present in supernatants of extracts of ovaries throughout the development of the ovary. However, the immunohistochemical study showed that there are marked seasonal changes in the distribution of FSG in ovaries. The polyclonal antibody reacted strongly with globules of accessory cells before the beginning of the breeding season (August to December). During the breeding season (February to April), the immunohistochemical reaction was found on the surface of oocytes but was weak in the accessory cells. At the ultrastructural level, the antibody reacted with globules of variable density in accessory cells. Intense immunolabelling was observed in the vacuole-like structures of the globules. Sometimes, products of the specific immunocytochemical reaction were found in the Golgi apparatus in these globules. Quantitative examination indicated that FSG was actively produced by the accessory cells from the late non-breeding season to the pre-breeding season. These results suggest that there are marked seasonal variations in the production of FSG by the accessory cells in the sea urchin ovary. These findings also provide new evidence that accessory cells exhibit dynamic changes during the reproductive process in the sea urchin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00188689
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  • 2
    Electronic Resource
    Electronic Resource
    The egg-jelly macromolecule, a fucose sulphate glycoconjugate, originates from the accessory cells of the ovary in the sea urchin Hemicentrotus pulcherrimus (1992)
    Springer
    Development genes and evolution 201 (1992), S. 179-189 
    Details
    ISSN: 1432-041X
    Keywords: Sea urchin ; Jelly coat ; Accessory cell ; Oogenesis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The immunocytochemical localization of the egg-jelly macromolecule, a fucose sulphate glycoconjugate (FSG) that induces the acrosome reaction in spermatozoa, was investigated in ovaries of the sea urchin Hemicentrotus pulcherrimus by use of a polyclonal antibody. The polyclonal antibody reacted with the accessory cells and oocytes in the ovarian lumen. In the accessory cells, evidence of an intense immunohistochemical reaction was observed in many globules of variable density. Products of the specific immunohistochemical reaction were frequently observed in the surface region of oocytes, at a distance from the ovarian wall. At the ultrastructural level, the polyclonal antibody was found to react with the material present in the vacuole-like structures of the globules in the accessory cells. Many gold particles, demonstrating specific immunolabelling, were associated with well-developed microvilli on the vitellogenic oocytes. In the mature oocytes, intense labelling was observed in the jelly coat but not in the vitelline coat. By contrast, oogonia and early oocytes were barely labelled. Quantitative data indicated that the extent of immunolabellings in the surface region of oocytes was very high in the vitellogenic and mature oocytes. In all cases, neither the oocyte cytoplasm nor the subcellular organelles were labelled. These results suggest that FSG is produced by the accessory cells and is deposited initially on the surface of vitellogenic oocytes for the formation of jelly. These findings may provide a new insight into the role of the accessory cells in the reproductive process of the sea urchin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00188717
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  • 3
    Electronic Resource
    Electronic Resource
    Occurrence and expression of acid phosphatase of Hymenoscyphus ericae (Read) Korf & Kernan, in isolation or associated with plant roots (1992)
    Springer
    Mycorrhiza 1 (1992), S. 137-146 
    Details
    ISSN: 1432-1890
    Keywords: Ericaceae ; Mycorrhizal fungi ; Acid phosphatase ; Protein expression ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The activity of acid phosphatase produced in pure culture by the endomycorrhizal fungus Hymenoscyphus ericae (Read) Korf & Kernan (H. ericae LPA 2) was inhibited by high phosphorus levels, alkaline pH, fluoride, molybdate and mannosidase, and activated by concanavalin A. Over 80% of the enzyme activity was due to two wall-bound acid phosphatase isozymes with the characteristics of mannose-rich glycoproteins. Antiserum was raised against the major, low-molecular-weight wall isozyme and its activity tested by immunoblotting and ELISA. The antiserum cross reacted 100% with exocellular (excreted) and 28% with cytoplasmic cellular fractions of H. ericae (LPA 2) cultures, and showed high reactivity with other strains of H. ericae but not with fungal isolates from Erica hispidula L. or E. mauritanica L. Ultrastructural localization of acid phosphatase by cytoenzymology and indirect immunogold labelling confirmed its association with the fungal wall in pure culture and showed that the influence of a high phosphorus level, fluoride and molybdate is through inactivation of the enzyme. Intense acid phosphatase activity, sensitive to the latter inhibitors, was also present on external hyphae growing over a host or non-host root but it was weak or absent from intracellular hyphae where these developed within a host root. Indirect immunolabelling confirmed that this acid phosphatase was of fungal origin and that the specific inhibitory effect of host cells is due to inactivation of the enzyme rather than repression of its synthesis. Possible implications of fungal acid phosphatase in ericoid endomycorrhizal infection processes are discussed together with mechanisms that may be regulating the enzyme activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00203287
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  • 4
    Electronic Resource
    Electronic Resource
    The AQP2 water channel: Effect of vasopressin treatment, microtubule disruption, and distribution in neonatal rats (1995)
    Springer
    The journal of membrane biology 143 (1995), S. 165-175 
    Details
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233445
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  • 5
    Electronic Resource
    Electronic Resource
    Expression of growth hormone receptor by immunocytochemistry in rat molar root formation and alveolar bone remodeling (1992)
    Springer
    Calcified tissue international 50 (1992), S. 541-546 
    Details
    ISSN: 1432-0827
    Keywords: Growth hormone ; Growth hormone receptor ; Odontogenesis ; Bone remodeling ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Growth hormone (GH) may regulate tooth formation and bone remodeling associated with tooth eruption. This study reports the distribution of growth hormone receptor/binding protein in developing rat molars and adjacent alveolar bone by immunocytochemistry using well-characterized anti-growth hormone receptor monoclonal antibodies. These tissues represent an excellent model for studying the ontogenic changes that occur in odontogenic and osteogenic cells, as these cells are found in linear arrays displaying the various stages of morphological and functional differention, and differentiated function. Immunoreactivity was first seen in precementoblasts in contact with the epithelial root sheath, and preodontoblasts. However, growth hormone receptor immunoreactivity was associated primarily with the cytoplasm of odontogenic and osteogenic cells forming their respective matrices. Thus, cementoblasts and odontoblasts at sites of new matrix formation showed intense immunoreactivity whereas cementocytes and mature odontoblasts at later stages of tooth development were nonreactive. Osteoblasts engaged in intramembranous ossification in the alveolar bone were positive, although osteocytes and endosteal cells were immunonegative. Osteoclasts at sites of alveolar bone remodeling resorption were also immunopositive. These patterns of receptor expression parallel the ontogenic sequences of odontogenic and osteogenic cells and suggest that GH promotes the functional state of these cells. Our results also imply that GH may influence differentiation or differentiated functions associated with odontogenesis, osteogenesis, and bone remodeling independent of systemic insulin-like GF-I.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00582170
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  • 6
    Electronic Resource
    Electronic Resource
    Periplasmic location of nitrous oxide reductase and its apoform in denitrifying Pseudomonas stutzeri (1992)
    Springer
    Archives of microbiology 157 (1992), S. 218-222 
    Details
    ISSN: 1432-072X
    Keywords: Denitrification ; N2O reductase ; Nitrite reductase ; Immunocytochemistry ; Localization ; Two-dimensional electrophoresis ; Cell fractionation ; Pseudomonas stutzeri
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunogold labelling techniques on ultrathin sections of low temperature embedded cells yielded evidence for the periplasmic location of the respiratory enzymes N2O reductase and nitrite reductase (cytochrome cd 1) in Pseudomonas stutzeri strain ZoBell. Cell fractionation by spheroplast preparation and two-dimensional electrophoresis showed the absence of a membrane association of these enzymes. Immunocytochemical localization of N2O reductase in a mutant strain deficient in the chromophore of N2O reductase showed the gold label at the cell periphery, indicating that the copper chromophore processing takes place after export of this protein's apoform.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00245153
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  • 7
    Electronic Resource
    Electronic Resource
    Cambium: old challenges – new opportunities (1999)
    Springer
    Trees 13 (1999), S. 138-151 
    Details
    ISSN: 0931-1890
    Keywords: Key words Cytoskeleton ; Immunocytochemistry ; Model systems ; Populus ; Secondary vascular system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Trees represent a, probably the, major component of the biosphere and have a unique place in the history of Mankind. One of their most fascinating features is the process of secondary growth which is effected principally by the secondary vascular system, the developmental continuum of secondary phloem, vascular cambium, and secondary xylem. However, for too long assumptions about the developmental biology of trees have had to be based upon studies of primary growth systems within annual, herbaceous species because study of the secondary vascular system had been largely ignored. Even when attempts are made to understand some of the most fundamental features of the secondary vascular system, such as xylogenesis, the current model system, isolated Zinnia mesophyll cells, is not entirely appropriate to the situation in the intact tree. Some deficiencies of the Zinnia system are discussed, and the advantages of the genus Populus as a model for study of the hardwood secondary vascular system are considered. Some of the new approaches which are poised to lead to significant advances in our knowledge of the cell bio-logy of the secondary vascular system of trees – spe-cifically of the cell wall, the plasmalemma, and the cytoskeleton – are discussed. The value of one of these new techniques – immunocytochemistry – is demonstrated by a consideration of recent work on the role of the cytoskeleton in the hardwood secondary vascular system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00009745
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  • 8
    Electronic Resource
    Electronic Resource
    Immunocytochemical identification of androgen receptor in mouse osteoclast-like multinucleated cells (1994)
    Springer
    Calcified tissue international 54 (1994), S. 325-326 
    Details
    ISSN: 1432-0827
    Keywords: Androgen Receptor ; Osteoclast ; Mouse ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Expression of androgen receptor (AR) in mouse osteoclast-like multi-nucleated cells (OCs) was examined with immunocytochemical techniques. Murine OCs were obtained by co-culturing mouse osteoblastic cells and bone marrow cells. Three preparations of polyclonal anti-AR antibody which were raised in rabbit against different parts of the human AR were employed for the experiments. Specific staining for AR was demonstrated in the nuclei and the perinuclear area of mouse OCs. This is the first report demonstrating the presence of AR in osteoclast-like cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00295958
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  • 9
    Electronic Resource
    Electronic Resource
    Peptidergic motoneurons in the buccal ganglia of Aplysia californica Immunocytochemical, morphological, and physiological characterizations (1991)
    Springer
    Journal of comparative physiology 168 (1991), S. 323-336 
    Details
    ISSN: 1432-1351
    Keywords: Aplysia ; Motoneurons ; Immunocytochemistry ; Small cardioactive peptides ; Facilitation ; Depression ; Buccal ; Feeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We used physiological recordings, intracellular dye injections and immunocytochemistry to further identify and characterize neurons in the buccal ganglia of Aplysia calif ornica expressing Small Cardioactive Peptide-like immunoreactivity (SCP-LI). Neurons were identified based upon soma size and position, input from premotor cells B4 and B5, axonal projections, muscle innervation patterns, and neuromuscular synaptic properties. SCP-LI was observed in several large ventral neurons including B6, B7, B9, B10, and B11, groups of s1 and s2 cluster cells, at least one cell located at a branch point of buccal nerve n2, and the previously characterized neurons B1, B2 and B15. B6, B7, B9, B10 and B11 are motoneurons to intrinsic muscles of the buccal mass, each displaying a unique innervation pattern and neuromuscular plasticity. Combined, these motoneurons innervate all major intrinsic buccal muscles (I1/I3, I2, I4, I5, I6). Correspondingly, SCP-LI processes were observed on all of these muscles. Innervation of multiple nonhomologous buccal muscles by individual motoneurons having extremely plastic neuromuscular synapses, represents a unique form of neuromuscular organization which is prevalent in this system. Our results show numerous SCPergic buccal motoneurons with widespread ganglionic processes and buccal muscle innervation, and support extensive use of SCPs in the control of feeding musculature.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00198352
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  • 10
    Electronic Resource
    Electronic Resource
    Circadian photoreception in the retinally degenerate mouse (rd/rd) (1991)
    Springer
    Journal of comparative physiology 169 (1991), S. 39-50 
    Details
    ISSN: 1432-1351
    Keywords: Photoreception ; Retinally degenerate ; Mouse ; Circadian ; Rods ; Cones ; 11-cis retinaldehyde ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have examined the effects of light on circadian locomotor rhythms in retinally degenerate mice (C57BL/6J mice homozygous for the rd allele: rd/rd). The sensitivity of circadian photoreception in these mice was determined by varying the irradiance of a 15 min light pulse (515 nm) given at circadian time 16 and meauring the magnitude of the phase shift of the locomotor rhythm. Experiments were performed on animals 80 days of age. Despite the loss of visual photoreceptors in the rd/rd retina, animals showed circadian responses to light that were indistinguishable from mice with normal retinas (rd/+ and +/+). While no photoreceptor outersegments were identified in the retina of rd/rd animals (80–100 days of age), we did identify a small number of perikarya that were immunoreactive for cone opsins, and even fewer cells that contained rod opsin. Using HPLC, we demonstrated the presence and photoisomerization of the rhodopsin chromophore 11-cis retinaldehyde. The rd/rd retinas contained about 2% of 11-cis retinaldehyde found in +/+ retinas. We have yet to determine whether the opsin immunoreactive perikarya or some other unidentified cell type mediate circadian light detection in the rd/rd retina.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00198171
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  • 11
    Electronic Resource
    Electronic Resource
    Opsin localization and chromophore retinoids identified within the basal brain of the lizard Anolis carolinensis (1993)
    Springer
    Journal of comparative physiology 172 (1993), S. 33-45 
    Details
    ISSN: 1432-1351
    Keywords: Photoreception ; Extraretinal Photoreceptor ; Chromophore ; Opsin ; Reptile ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Since the beginning of this century evidence has accumulated which demonstrates that non-mammalian vertebrates possess photoreceptors situated deep within the brain. While many attempts have been made to localize these sensory cells, studies have either failed or been inconclusive. In this report we have used several experimental approaches to localize the deep brain photoreceptors of the lizard Anolis carolinensis. Using 3 antibodies that bind vertebrate cone opsins, we have immunolabelled cerebrospinal fluid (CSF)-contacting neurons located at the ventricular border within the nucleus ventromedialis of the septum. Western blot analysis indicates that these antibodies recognized a single 40 kD protein in ocular, anterior brain, and pineal extracts. Immunoblots of rodent brain did not show a similar protein band. We have also identified specific retinoids associated with phototransduction (11-cis and all-trans-3,4-didehydroretinaldehyde) within anterior brain extracts. This combined data provides the most detailed analysis of deep brain photoreceptors in any vertebrate. Consequently, we feel Anolis provides an excellent model to study this unexplored sensory system of the vertebrates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00214713
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  • 12
    Electronic Resource
    Electronic Resource
    Isolation of an egg-laying hormone-binding protein from the gonad of Aplysia californica and its localization in oocytes (1993)
    Springer
    Journal of comparative physiology 173 (1993), S. 475-483 
    Details
    ISSN: 1432-1351
    Keywords: Egg laying hormone ; Aplysia ; Binding protein ; Immunocytochemistry ; Reproductive system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A protein solubilized from a membrane preparation of the gonad of Aplysia californica has been isolated by affinity chromatography, using bag cell egg-laying hormone (ELH) as the bound ligand, and partially purified and characterized by gel electrophoresis. The protein has an apparent molecular weight of 52 kDa and consists of two disulfide-linked subunits of about 30 kDa each. The protein is glycosylated and has an acidic pI. Approximately 10–15 μg of this protein can be isolated from a single ovotestis, representing less than 1% of the total protein in the gonad; but the protein could not be detected in buccal mass or body wall, tissues which do not have apparent response to ELH. Antibodies generated against this ELH-binding protein (ELHBP) were used to localize sites in the ovotestis which might contain this molecule and thus represent targets for egg-laying hormone. Immunocytochemical results indicate that the oocytes are a rich source of this protein, since their cytoplasm was the only detectable site of immunoreactivity. Whether this binding protein represents an egg-laying hormone receptor is uncertain, but its prevalence in oocytes suggests that ELH plays a signaling role on these gametes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193520
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  • 13
    Electronic Resource
    Electronic Resource
    Immunogold silver staining for light microscopy (1996)
    Springer
    Histochemistry and cell biology 106 (1996), S. 9-17 
    Details
    ISSN: 1432-119X
    Keywords: Key words Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004180050019
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  • 14
    Electronic Resource
    Electronic Resource
    Immunogold silver staining for light microscopy (1996)
    Springer
    Histochemistry and cell biology 106 (1996), S. 9-17 
    Details
    ISSN: 1432-119X
    Keywords: Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02473198
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  • 15
    Electronic Resource
    Electronic Resource
    Subcellular localization of β-1,3-glucanase in Puccinia recondita f.sp. tritici-infected wheat leaves (1998)
    Springer
    Planta 204 (1998), S. 324-334 
    Details
    ISSN: 1432-2048
    Keywords: Key words:β-1 ; 3-Glucanase ; Immunocytochemistry ; Leaf rust pathogen ; Resistance ; Triticum (pathogen resistance)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. An antiserum raised against the purified 33-kDa β-1,3-glucanase of wheat (Triticum aestivum L.) was employed to investigate the ultrastructural localization of the enzyme in wheat leaves infected with Puccinia recondita Rob. ex Desm. f.sp. tritici Eriks. and Henn. using a post-embedding immunogold labelling technique. In both compatible and incompatible interactions, β-1,3-glucanase was detected in the host plasmalemma and in the domain of the host cell wall near the plasmalemma of the mesophyll cells, but higher concentrations of the enzyme were detected in infected resistant wheat leaves than in infected susceptible ones. β-1,3-Glucanase was also found in the secondary thickening of xylem vessels and in the walls of guard cells, epidermal cells and phloem elements, while no labelling was observed in host organelles, viz. vacuoles, mitochondria, endoplasmic reticulum, Golgi bodies, nuclei and chloroplasts. A low concentration of the enzyme was detected on the intercellular hyphal wall and in the hyphal cytoplasm. In the compatible interaction, β-1,3-glucanase was demonstrated to accumulate predominantly in the haustorial wall and extrahaustorial matrix. In the incompatible interaction, strong labelling for β-1,3-glucanase was found in host cell wall appositions, in the extracellular matrix in the intercellular space, and in electron-dense structures of host origin which occurred in the incompatible interaction only.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004250050263
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  • 16
    Electronic Resource
    Electronic Resource
    Microfilaments (F-actin) in generative cells and sperm: an evaluation (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 89-100 
    Details
    ISSN: 1432-2145
    Keywords: Actin ; Cytoskeleton ; Generative cell ; Immunocytochemistry ; Microtubule ; Mitosis ; Phragmoplast ; Pollen ; Rhodamine phalloidin ; Sperm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Disagreement has arisen over the presence of actin-containing microfilaments (Mfs) in angiosperm generative cells and sperm (GSP). In order to address this issue, we subjected GSP of Tradescantia virginiana, Nicotiana tabacum and Rhododendron laetum to a series of localizations using different antiactins, rhodamine phalloidin and antimyosin. Coordinate staining with antitubulin and Hoechst 33258 defined the status of the microtubule (Mt) cytoskeleton and stages of generative cell division. Additional experiments utilized cytochalasin D (CD). In no instance could Mfs be detected in GSP of the three species. Instead, Mfs seen at the periphery of GSP appear to be continuous with vegetative Mfs and thus are in the vegetative cytoplasm. Mfs are not seen in the constriction zone of dividing T. virginiana generative cells, nor are they indicated in the phragmoplast of N. tabacum and R. laetum. Myosin localizations reveal punctate staining in the vegetative cytoplasm and a thin line of fluorescence around the the outside of the generative cell. While CD seems to delay generative cell division, cytokinesis still takes place. CD-induced Mf fragments are evident in the vegetative cytoplasm but not in GSP. The weight of evidence therefore indicates that GSP do not contain Mfs. The implications of this conclusion for the behavior of GSP and the mechanism of cytokinesis in dividing generative cells are considerable.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00194867
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  • 17
    Electronic Resource
    Electronic Resource
    Immunocytochemical study of cell cycle control by cytokinin in cultured soybean cells (1996)
    Springer
    Journal of plant growth regulation 15 (1996), S. 95-102 
    Details
    ISSN: 1435-8107
    Keywords: Cell cycle ; Cytokinin ; Soybean cells ; Glycine max ; Immunocytochemistry ; S phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract An immunocytochemical method was used to determine the proportion of cells in the DNA synthesis (S phase) of the mitotic cycle in suspension cultures of soybean (Glycine max (L.) Merr. cv. Acme) callus of cotyledonary origin, the stably cytokinin-dependent tissue used in the cytokinin bioassay devised by Carlos O. Miller. A standard cell synchronization protocol involving hydroxyurea was used to demonstrate the applicability of the immunocytochemical method to this cell culture. Cells were brought to mitotic arrest by cytokinin withdrawal, and the cell division cycle was restarted by the addition of cytokinin. We have followed the pattern of resumption of S phase after the readdition of cytokinin. This pattern reveals the existence of three subpopulations of cells in cytokinin-starved cultures, consistent with the occurrence of three cytokinin-requiring events in the cell cycle: one in mitosis, one in S phase, and one in the G1 phase.
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    URL: http://dx.doi.org/10.1007/BF00192938
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    Carbon dioxide laser induction of heat shock protein 70 synthesis: Comparison with high temperature treatment (1995)
    Springer
    Lasers in medical science 10 (1995), S. 207-212 
    Details
    ISSN: 1435-604X
    Keywords: Heat shock protein 70 (hsp70) ; Carbon dioxide laser ; Sodium arsenite ; Human diploid fibroblast ; Electrophoresis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Physics , Technology
    Notes: Abstract In a previous study, it was demonstrated that heat shock protein 70 (hsp70) was induced by short duration (〈1 s) carbon dioxide (CO2) laser radiation (10.6Μm. To further characterize the stress response after laser irradiation, the time course of synthesis and cellular localization of hsp70 has been followed. As it had been shown that laser irradiation elevated the temperature to about 67±2
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    URL: http://dx.doi.org/10.1007/BF02133333
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    Gonadotropin-releasing hormone (GnRH) pathways and reproductive control in elasmobranchs (1993)
    Springer
    Environmental biology of fishes 38 (1993), S. 209-218 
    Details
    ISSN: 1573-5133
    Keywords: Terminal nerve ; Midbrain ; Immunocytochemistry ; Reproduction ; Brain ; Sharks ; Rays ; Skates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis Immunoreactive (ir) gonadotropin-releasing hormone (GnRH) is localized in many neurons of the terminal nerve (TN) and midbrain tegmentum, while few ir-cells are observed in the preoptic area and ventral hypothalamus. The paucity of preoptic ir-cells may relate to an unusual feature of the elasmobranch pituitary, i.e. a lack of portal control of gonadotropin-producing cells. TN and midbrain GnRH-ir neurons may be major sources of GnRH used to modulate or otherwise control both pituitary and brain cells via delivery through the systemic circulation. These ir-nuclei also appear to directly innervate CNS regions (the preoptic area, habenula and clasper control area of the spinal cord) involved in sexual functions. Important regulatory mechanisms, represented by interactions between GnRH pathways and sex-steroid concentrating neurons, are likely to occur in the preoptic area, habenula and midbrain tegmentum.
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    URL: http://dx.doi.org/10.1007/BF00842917
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  • 20
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    Effects of aflatoxin B1 on in vitro cultures ofNicotiana tabacum var. Samsun (1994)
    Springer
    Mycopathologia 125 (1994), S. 107-117 
    Details
    ISSN: 1573-0832
    Keywords: Aflatoxin B1 ; Immunocytochemistry ; Regeneration ; Tissue culture ; Tobacco plantlets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effects of aflatoxin B1, (0.5–25 µg ml−1) on in vitro root and shoot development in young tobacco explants were investigated. Despite an initial apparent stimulatory effect on most measured parameters at 0.5 µg ml−1 AFB1, the number of leaves, root and leaf mass per plantlet were progressively inhibited with increasing AFB1 concentration. The number of explants developing roots was reduced to 34% at the highest (25 µg ml−1) AFB1 concentration, following 3 weeks exposure to the toxin. Leaf chlorophyll content at this toxin concentration was significantly lower than that measured for control plantlets. Thin layer chromatography confirmed the absorption of AFB1 by the plantlets. Using immunocytochemical techniques, AFB1 was immunolocated predominantly in the vacuoles, the nucleus and the cytoplasm (possibly intravesicularly). The results are discussed in terms of this immunolocation within the cell.
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    URL: http://dx.doi.org/10.1007/BF01371099
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  • 21
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    Infection and disintegration of vascular tissues of non-suberized roots of spruce byHeterobasidion annosum and use of antibodies for characterizing infection (1995)
    Springer
    Mycopathologia 129 (1995), S. 91-101 
    Details
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
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    URL: http://dx.doi.org/10.1007/BF01103468
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  • 22
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    Double labeling of mRNA and protein markers in cultured embryoid bodies (1994)
    Springer
    Methods in cell science 16 (1994), S. 11-16 
    Details
    ISSN: 1573-0603
    Keywords: Differentiation ; DIG-mRNA ; Embryoid bodies ; ES cell ; Immunocytochemistry ; In situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In vitro suspension cultures of embryonal carcinoma or embryonic stem cells (EC/ES) generate cell aggregates termed as embryoid bodies (EBs). EBs have been analyzed to study the mechanisms of cellular differentiation in vitro. The multipotency of EC/ES cells to differentiate into various cell types as well as the expression of many marker genes provides a valuable in vitro model system to study the mechanisms of cellular differentiation. Here we present a procedure for a mRNA detection of a specific gene using double labeling-mRNA probe and an antibody against cellular marker proteins. This double labeling analysis in combination with a culture of EBs provides a useful approach to analyze several mechanisms of cellular differentiation from multipotent EC/ES cells.
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    URL: http://dx.doi.org/10.1007/BF01404831
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  • 23
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    The immunocytochemical localization of angiotensinogen in the rat ovary (1990)
    Springer
    Cell & tissue research 261 (1990), S. 367-373 
    Details
    ISSN: 1432-0878
    Keywords: Angiotensinogen ; Ovary ; Estrous cycle Renin-angiotensin system ; Atresia ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study examined the presence and cellular distribution of angiotensinogen, the precursor to the angiotensin peptides, in the ovary of the normal cycling rat by immunocytochemistry. Angiotensinogen staining was present in the granulosa cells of maturing follicles and to a lesser extent in those undergoing atresia. Staining was not seen in the granulosa cells of primordial or early primary follicles. In maturing follicles intense staining for angiotensinogen was confined to the antral cell layers, cells of the cumulus oophorus and in the follicular fluid. Strong immunostaining was also seen in the germinal epithelium covering the ovary. Lighter angiotensinogen staining was observed in some parts of the cortical and medullary stroma and occasionally in corpora lutea. No variation in the intensity or pattern of angiotensinogen staining was observed throughout the estrous cycle. Comparison of the distribution of angiotensinogen with the previously described localization of renin, AII, angiotensin converting enzyme and AII receptors, suggests that there are a number of intra-ovarian sites at which AII could be produced.
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    URL: http://dx.doi.org/10.1007/BF00318679
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    An immunogold, cryoultrastructural study of sites of synthesis and storage of chorionic gonadotropin and placental lactogen in human syncytiotrophoblast (1990)
    Springer
    Cell & tissue research 261 (1990), S. 375-382 
    Details
    ISSN: 1432-0878
    Keywords: Chorionic gonadotropin ; Placental lactogen Placenta ; Cryoultrastructure ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The sites of intracellular synthesis and storage of human placental lactogen (hPL) and human chorionic gonadotropin (hCG) are controversial. We have used one of the most sensitive methods, cryoultramicrotomy and immunogold labelling, to localise these hormones at the electron-microscopic level. In both 12-week and term placentas hCG and hPL are present throughout the rough endoplasmic reticulum cisternae, in the Golgi bodies, and in the infrequent small dense granules of the syncytiotrophoblast. Previous assays have shown that hCG is at a higher concentration in early pregnancy and hPL peaks in late pregnancy, and our results corroborate these findings. No significant localisation of either hormone was seen in the cytotrophoblast or villous stroma. The results suggest that both hCG and hPL are synthesised and packaged by the classical secretory pathway, although the level of hormone stored in granules at any one time is small.
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    URL: http://dx.doi.org/10.1007/BF00318680
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    Developmental expression of transforming growth factor-α and epidermal growth factor receptor proteins in the human pancreas and digestive tract (1994)
    Springer
    Cell & tissue research 278 (1994), S. 439-450 
    Details
    ISSN: 1432-0878
    Keywords: Transforming growth factor alpha ; Epidermal growth factor receptor ; Development, ontogenetic ; Digestive tract ; Endocrine cells ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This study was designed to localize transforming growth factor alpha (TGF-α) and epidermal growth factor receptor (EGFR) expression in the developing human gastrointestinal tract and pancreas. Immunohistochemical techniques using specific antibodies against human TGF-α and EGFR were performed on digestive tissues of fetuses from 9 to 10 to 24 weeks of gestation, children and adults. In fetuses, TGF-α and EGFR proteins were expressed in all epithelial tissues studied with a good correlation and from an age as early as 9 to 10 weeks of gestation, except for TGF-α in the esophagus. The strongest TGF-α immunostaining was noted in the stomach and the proximal colon. Unexpectedly, immunoreactive gut endocrine cells were observed with the two antibodies used. Relatively numerous in fetuses, they decreased in number with age and were rare in adults particularly along the colon. Enteroglucagon-secreting cells were shown to express TGF-α while some gastrin, somatostatin and pancreatic glucagon cells were immunostained with EGFR antibodies. The presence of TGF-α and of its recetor in digestive tract epithelium and pancreatic tissues early in fetal life suggests a functional role for TGF-α during the developmental process of the digestive system. We demonstrate that TGF-α is also produced by endocrine cells and might have an additional mode of action other than paracrine, at least during fetal life.
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    URL: http://dx.doi.org/10.1007/BF00331362
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  • 26
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    Immunoreactivity to the pancreatic polypeptide family in the nervous system of the adult human blood fluke, Schistosoma mansoni (1990)
    Springer
    Cell & tissue research 261 (1990), S. 573-581 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptides (pancreatic polypeptide, peptide YY, neuropeptide Y) ; Immunocytochemistry ; Confocal scanning laser microscopy ; Schistosoma mansoni (Scolecida, Trematoda)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and distribution of neuropeptides belonging to the pancreatic polypeptide family have been demonstrated by an indirect immunofluorescence technique in the nervous systems of adult male and female Schistosoma mansoni. Seven antisera of differing regional specificity to pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY) were employed on both whole-mount and cryostat-sectioned material. Positive immunoreactivity (IR) was obtained with all antisera except an N-terminally-directed antiserum to NPY. In the CNS, immunoreactivity was restricted to cell bodies and nerve fibres in the anterior ganglia, central commissure and dorsal and ventral nerve cords of both sexes, whereas, in the PNS, positive-IR was present in the plexuses innervating the subtegumental musculature and the oral and ventral suckers. Intense immunoreactivity was observed in a plexus of nerve fibres and cell bodies in the lining of the gynaecophoric canal and in fine nerve fibres innervating the dorsal tubercles of the male. In contrast, in the female, strong immunoreactivity was evident in nerve plexuses innervating the lining of the ovovitelline duct and in the wall of the ootype, but most notably in a cluster of cells in the region of Mehlis' gland. Results suggest that molecules with C-terminal homology to the PP-family are present in S. mansoni. These peptides would appear to be important regulatory molecules in the parasite's nervous system and may play a role in the control of egg production.
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    URL: http://dx.doi.org/10.1007/BF00313537
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  • 27
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    Distribution and functional significance of Met-enkephalin-Arg6-Phe7-and Met-enkephalin-Arg6-Gly7-Leu8-like peptides in the blowflyCalliphora vomitoria (1990)
    Springer
    Cell & tissue research 259 (1990), S. 147-157 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Met-enkephalin-Arg6-Phe7 ; Met-enkephalin-Arg6-Gly7-Leu8 ; Neurosecretory cells of insects ; Neuropeptides ; Co-existence of peptides ; Blowfly,Calliphora vomitoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal pathways in the retrocerebral complex and thoracico-abdominal ganglionic mass of the blowflyCalliphora vomitoria have been identified immunocytochemically with antisera against the extended-enkephalins, Met-enkephalin-Arg6-Phe7 (Met-7) and Met-enkephalin-Arg6-Gly7-Leu8 (Met-8). Neurons of the hypocerebral ganglion, immunoreactive to Met-8, have axons in the crop duct nerve and terminals in muscles of the crop and its duct. Certain neurons of the hypocerebral ganglion are also immunoreactive to Met-7, and axons from these cells innervate the heart. Met-8 immunoreactive nerve terminals invest the cells of the corpus allatum. The source of this material is believed to ve a single pair of lateral neurosecretory cells in the brain. There is no Met-7 immunoreactive material in the corpus allatum. In the corpus cardiacum neither Met-7 nor Met-8 immunoreactivity is present in the cells. However, in the neuropil of the gland certain fibres, with their origins elsewhere, do contain Met-8 immunoreactivity. The most prominent neurons in the thoracic ganglion are the Met-7 immunoreactive ventral thoracic neurosecretory cells, axons from which project to neurohaemal areas in the dorsal neural sheath and also, via the ventral connective, to the brain. Co-localisation studies show that the perikarya of these cells are immunoreactive to antisera raised against several vertebrate-type peptides, such as Met-7, gastrin/cholecystokinin and pancreatic polypeptide. However, their axons and terminals show varying amounts of the peptides, suggesting differential transport and utilisation. Only a few cells in the thoracic ganglion are immunoreactive to Met-8 antisera. These lie close to the nerve bundles suppling the legs. In the abdominal ganglion, Met-8 immunoreactive neurons project to the muscles of the hindgut. This study suggests that the extended enkephalin-like peptides ofCalliphora may have a variety of different roles: as neurotransmitter or neuromodulator substances; in the direct innervation of effector organs; and as neurohormones.
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    URL: http://dx.doi.org/10.1007/BF00571439
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    Immunocytochemical evidence of a met-enkephalin-like substance in the dense-core granules of mouse Merkel cells (1991)
    Springer
    Cell & tissue research 265 (1991), S. 611-614 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Met-enkephalin ; Merkel cells ; Dense-core granules ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The electron-microscopic immunogold method was applied to Merkel cells of adult mice to demonstrate the subcellular localization of met-enkephalin-like immunoreactivity. Post-embedding incubation with metenkephalin antisera showed that the gold particles were associated with the dense-core granules of the Merkel cells. The majority, but not all, of the dense-core granules were strongly labelled. Osmication caused a significant reduction in the number of gold particles on these granules. The nerve terminal associated with the Merkel cell did not show met-enkephalin-like immunoreactivity. To the best of our knowledge, this is the first report of the ultrastructural localization of a positive met-enkephalin immunoreactivity in the dense-core granules of Merkel cells in mice.
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    URL: http://dx.doi.org/10.1007/BF00340885
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    Presence of an oxytocin-like peptide in the hypothalamus and neurohypophysis of a turtle (Mauremys caspica) and a snake (Natrix maura) (1994)
    Springer
    Cell & tissue research 279 (1994), S. 75-84 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin different from the mammalian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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    URL: http://dx.doi.org/10.1007/s004410050263
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    The biased intracellular accumulation of the β-subunit of salmon gonadotropin (GTH II) in the pituitary of rainbow trout Oncorhynchus mykiss, during gametogenesis (1994)
    Springer
    Cell & tissue research 279 (1994), S. 93-99 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ-labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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    URL: http://dx.doi.org/10.1007/s004410050265
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    Photoreceptor membrane turnover in the crayfish Cherax destructor: electron microscopy and anti-rhodopsin electron-microscopic immunocytochemistry (1990)
    Springer
    Cell & tissue research 262 (1990), S. 483-499 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Photoreceptor cells ; Rhodopsin ; Membrane recycling ; Cherax destructor (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Examination of the ultrastructure of retinula cells of the Australian crayfish Cherax destructor at different times over a 24-hour cycle, together with patterns of anti-rhodopsin antigenicity, has lead to the formulation of a model of photoreceptor membrane turnover in these animals. Its main features are: (a) the existence of two bursts of rhabdomeral membrane breakdown; one, light-sensitive and synchronous, occurring at dawn, the other, constituting the first part of the membrane replacement phase itself, occurring during the afternoon and night, (b) the desynchronisation of the replacement phase of turnover between animals and to a lesser extent between cells of the same retina, (c) confinement of ultrastructurally detectable signs of photoreceptor membrane processing to the retinula cells themselves, and (d) replacement of a substantial part if not all of the rhabdomeral membrane daily. This model is compatible with many of the observations reported on the American crayfish Procambarus, and utilises the same basic mechanisms that are believed to operate in photoreceptor membrane turnover in many other arthropod compound eyes.
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    URL: http://dx.doi.org/10.1007/BF00305244
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    Immunocytochemical localization of vasotocin-like immunoreactivity in the brain of the cartilaginous fish, Scyliorhinus caniculus (1990)
    Springer
    Cell & tissue research 262 (1990), S. 507-513 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasotocin ; Hypothalamus ; Neurosecretory fibers ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of vasotocin-like peptides in the central nervous system of the cartilaginous fish Scyliorhinus canicula was determined by indirect immunofluorescence and peroxidase anti-peroxidase techniques, using a specific antiserum raised in rabbits against synthetic vasotocin. Immunoreactive perikarya were mainly detected in the anterior hypothalamus, within the midcaudal part of the preoptic nucleus. The most rostral positive cell bodies were located in the dorso-lateral parts of the preoptic area, whereas at a more caudal level, they took a ventro-medial position within the deepest layers of the nucleus. Throughout the preoptic region these cells varied in shape according to their location. Occasionally, scattered vasotocin-like immunopositive cells were also identified in the nucleus periventricularis hypothalami. Vasotocin immunoreactivity was detected in numerous varicose nerve fibers of the preopticohypophysial tract. These fibers were seen to course through the medio-basal hypothalamus and caudally, after having passed the hypophysial stem, they reached the neurointermediate lobe of the pituitary. Numerous immunoreactive fibers were also observed within the rostro-medial region of the median eminence. At this level the fibers were in close proximity to the capillary loops. In the preoptic region, some stained cells exibited short processes that appeared to contact non-reactive perikarya. By comparing the distribution of vasotocin- and corticotropin-releasing factor immunoreactivity on adjacent then serial sections, it was revealed that these peptides, in S. canicula, do not coexist in the same perikarya. The present results, are compared with those obtained in other vertebrate groups, and their possible functional implications are discussed.
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    PYY-like material and its spatial relationship with NPY, CGRP and 5-HT in the lung of the Syrian golden hamster (1990)
    Springer
    Cell & tissue research 262 (1990), S. 543-550 
    Details
    ISSN: 1432-0878
    Keywords: PYY ; NPY ; CGRP ; Serotonin ; Lung ; Radioimmunoassay ; Immunocytochemistry ; Mesocricetus auratus (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We investigated the presence of peptide YY, neuropeptide Y, calcitonin gene-related peptide and serotonin in the hamster lung by radioimmunoassay, high performance liquid chromatography and immunocytochemistry. Lung-tissue concentrations of peptide YY and neuropeptide Y were 1.3±0.2 and 2.5±0.2 pmol/g wet weight, respectively. These two closely related pancreatic peptides were demonstrated in separate peaks with high performance liquid chromatography. The peptide YY appeared fragmented as immunoreactive peptide YY eluted primarily late in the gradient but showed additional peaks early in the gradient. Peptide YY-like immunoreactivity (PYY-LI) was predominantly observed in one or more cells of neuroepithelial bodies in all airways peripheral to bronchioles, and in solitary neuroendocrine cells primarily located in the same peripheral areas. Neuropeptide Y-LI was seen in individual, thin nerve fibers around arteries and veins, in the airway lamina propria, and in the airway epithelium; in the latter also immunopositive nerve terminals were located. This pattern did not appear to coincide with that of calcitonin gene-related peptide-LI in epithelial nerve fibers and terminals. Peptide YY-LI, calcitonin gene-related-LI and serotonin-LI were present in cells of one and the same neuroepithelial body. However, peptide YY-LI was never found to be co-localized with calcitonin gene-related-LI or serotonin-LI, but the latter two were co-localized as previously reported.
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    URL: http://dx.doi.org/10.1007/BF00305251
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
    Details
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    Cellular localization of somatolactin in the pars intermedia of some teleost fishes (1991)
    Springer
    Cell & tissue research 263 (1991), S. 207-215 
    Details
    ISSN: 1432-0878
    Keywords: Somatolactin (SL) ; Pituitary gland, pars intermedia ; PAS-positive cells ; Immunocytochemistry ; Gadus morhua, Oncorhynchus mykiss, Poecillia latipinna (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We report here on the cellular localization in the fish pituitary of somatolactin (SL), a putative new pituitary hormone related to growth hormone and prolactin, which has been recently identified in the piscine pituitary gland. Immunocytochemical staining, using anti-cod SL serum, revealed that in the cod pituitary gland, SL is produced by cells in the intermediate lobe, bordering the neural tissue. These cells, staining weakly with periodic-acid-Schiff (PAS), are distinct from the melanocyte stimulating hormone (MSH) cells which, as in all teleosts, are PAS-negative. SL-immunoreactivity was observed in the same location in all other teleost species examined: flounder, rainbow trout, killifish, molly, catfish and eel. In most fish the SL-immunoreactive cells are either strongly or weakly PAS-positive but in rainbow trout are chromophobic, indicating that the SL protein can probably exist in glycosylated and non-glycosylated forms. Thus, in demonstrating the cellular localization of SL, this study provides the first identification of the enigmatic, second cell-type of the fish pars intermedia.
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    URL: http://dx.doi.org/10.1007/BF00318762
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    Leucokinin-like immunoreactive neurones in the central nervous system of the spider Cupiennius salei (1996)
    Springer
    Cell & tissue research 284 (1996), S. 143-152 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Leucokinins ; Immunocytochemistry ; Nervous system ; central ; Neuromodulators ; Cupiennius salei (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Leucokinin is a member of the myokinin peptide family. These myotropic peptides are widely distributed in arthropods. A specific antiserum immunoreactive to the neuroactive octapeptide leucokinin I (LKI) has been used to map neurones within the central nervous system of the central American wandering spider Cupiennius salei. The antiserum labels nine pairs of cell bodies and their axons. The somata are grouped near the joint of the optic lobes in the dorsal part of the supraoesophageal ganglion. The axons descend in a bundle, pass the oesophagus and divide to innervate the suboesophageal ganglion at three levels. In the dorsal-most layer, five small parallel fibres travel within the medio-central tract towards the opisthosomal neuromeres and give off small varicose projections into all leg neuromeres. In the middle layer, two neurones in the sensory-longitudinal tract 3 project into each leg neuromere. In the ventral layer, the two largest fibres (about 8 μm) run in the medio-ventral tract, each giving off a branch into all leg neuromeres. These first order arborizations have small second order arborizations in the ventral part of the leg neuromere and ascend to the five dorsal fibres in the medio-ventral tract where they form varicosities. The second order arborizations form extensive varicosities in the ventral part of the leg neuromere. These neurones may play a role in the intercellular communication between the sensory input and the motor output system, because they are represented in the dorsal sensory and the ventral motor neuropile. The ventral fibres resemble intersegmental interneurones and may function as modulators for leg motor neurones.
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    URL: http://dx.doi.org/10.1007/s004410050574
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    Localization of hydroxyindole O-methyltransferase-synthesizing cells in bovine epithalamus: immunocytochemistry and in-situ hybridization (1991)
    Springer
    Cell & tissue research 263 (1991), S. 413-418 
    Details
    ISSN: 1432-0878
    Keywords: Pineal organ ; Epithalamus ; Pinealocytes ; Hydroxyindole O-methyltransferase (HIOMT) ; Immunocytochemistry ; HIOMT-mRNA ; In-situ hybridization ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Hydroxyindole O-methyltransferase (HIOMT)-immunoreactive cells and melatonin synthesis were demonstrated in bovine epithalamus (including the pineal gland) with monoclonal antibodies and cRNA probes to HIOMT. The HIOMT-immunoreactive product was present in the cytoplasm of pinealocytes. All identifiable pinealocytes were clearly labeled in the pineal gland. The expression of the HIOMT gene was first identified in pinealocyte cytoplasm by in-situ hybridization (ISH). The distribution of the hybridization-positive cells in the pineal gland was compatible with that revealed by immunocytochemistry using the monoclonal antibody to HIOMT. In addition, HIOMT transcripts were found in the medial habenular nucleus, and the habenular and posterior commissure; they may correspond to S-antigen-immunoreactive cells demonstrated in the same regions of the hamster and the mouse. In these regions, the hybridization-positive cells did not exhibit HIOMT-immunoreactivity; thus, cells devoid of immunoreactivity may synthesize but rapidly transport the newly synthesized proteins. These results indicate (1) that the conversion of N-acetylserotonin into melatonin takes place in the cytoplasm of pinealocytes, (2) that some epithalamic cells in the habenular area may synthesize melatonin, and (3) that melatonin may act as a chemical messenger in centrally directed processes, as shown by using S-antigen immunocytochemistry.
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    URL: http://dx.doi.org/10.1007/BF00327275
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    Light- and electron-microscopic immunocytochemistry of a molluscan insulin-related peptide in the central nervous system of Planorbarius corneus (1992)
    Springer
    Cell & tissue research 267 (1992), S. 473-481 
    Details
    ISSN: 1432-0878
    Keywords: Cerebral ganglia ; Neurohormones ; Molluscan insulin-related peptide ; Immunocytochemistry ; Tannic acid ; Planorbarius corneus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two groups of cerebral dorsal cells of the pulmonate snail Planorbarius corneus stain positively with antisera raised against synthetic fragments of the B- and C-chain of the molluscan pro-insulin-related prohormone, proMIP-I, of another pulmonate snail, Lymnaea stagnalis. At the light-microscopic level the somata of the dorsal cells and their axons and neurohemal axon terminals in the periphery of the paired median lip nerves are immunoreactive with both antisera. Furthermore, the canopy cells in the lateral lobes of the cerebral ganglia are positive. In addition, MIPB-immunoreactive neurons are found in most other ganglia of the central nervous system. At the ultrastructural level, pale and dark secretory granules are found in somata and axon terminals of the dorsal cells. Dark granules are about 4 times as immunoreactive to both antisera as pale granules. Release of anti-MIPB- and anti-MIPC-immunopositive contents of the secretory granules by exocytosis is apparent in material treated according to the tannic acid method. It is concluded that the dorsal and canopy cells synthesize a molluscan insulin-related peptide that is packed in the cell body into secretory granules and that is subsequently transported to the neurohemal axon terminals and released into the hemolymph by exocytosis. Thus, MIP seems to act as a neurohormone on peripheral targets. On the basis of the analogy between the dorsal cells and the MIP-producing cells in L. stagnalis, it is proposed that the dorsal cells of P. corneus are involved in the control of body growth and associated processes.
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    URL: http://dx.doi.org/10.1007/BF00319369
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    Culture and characterization of dental follicle cells from rat molars (1992)
    Springer
    Cell & tissue research 267 (1992), S. 483-492 
    Details
    ISSN: 1432-0878
    Keywords: Dental follicle ; Cell culture ; Fibroblasts ; Immunocytochemistry ; Ultrastructure ; Collagen ; Gel-electrophoresis ; Western blotting ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Because the dental follicle is necessary for the eruption of teeth of limited eruption, it was the objective of this study to determine if the cells of the follicle could be cultured in vitro. To achieve this, dental follicles and associated enamel organs were dissected from the first and second mandibular molars of 6–7-day-old rats (secretory stage of amelogenesis), and then cultured in a medium that promotes fibroblast growth — the predominant cell type of the dental follicle. The cultured cells grew to confluency and were kept through 3 passages before experimentation. The cultured cells were fibroblastic in shape, elongate with processes, and transmission electron microscopy revealed that they contained an abundant rough endoplasmic reticulum, but did not form desmosomes. Immunofluorescent staining for anti-vimentin showed that all the cells stained and electron-microscopic immunogold labeling indicated that the antibody was associated with intermediate filaments. As revealed by SDS-polyacrylamide gel electrophoresis and Western blotting, the cultured cells synthesized and secreted the extracellular matrix molecules fibronectin and procollagens. Subsequent immunofluorescence staining of permeabilized and non-permeabilized cells confirmed the presence of fibronectin and type I collagen both intra- and extracellularly. Thus, based on all the above characteristics, the cultured cells appeared to be fibroblasts derived from the dental follicle, although a few of the fibroblasts may be derived from undifferentiated mesenchymal cells interposed between the alveolar bone and follicle. Experiments now can be conducted to determine how these cultured cells respond directly to growth factors that alter the rates of tooth eruption.
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    Synaptic contacts of tyrosine hydroxylase-immunoreactive elements in the inner plexiform layer of the retina of Bufo marinus (1992)
    Springer
    Cell & tissue research 267 (1992), S. 525-534 
    Details
    ISSN: 1432-0878
    Keywords: Retina ; Dopaminergic neurons ; Synapses ; Inner plexiform layer ; Immunocytochemistry ; Electron microscopy ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Tyrosine hydroxylase (TH) immunocytochemistry was utilized to quantify dopaminergic synapses in the inner plexiform layer of the retina of Bufo marinus. Since dopaminergic cells have bistratified dendritic arborisation in the inner plexiform layer, attention was given to the segregation of synapses between the scleral and the vitreal sublaminae. Light-microscopically, a more elaborate dendritic branching was observed in the scleral than in the vitreal sublamina. In contrast, about 55% of synapses occurred in the vitreal one fifth of the inner plexiform layer, 30% in the scleral fifth, and 15% in the intermediate laminae. Input sources and output targets showed only minor quantitative differences between sublaminae 1 and 5. TH-immunoreactive processes were found in presynaptic (62.8%) and postsynaptic (37.2%) positions. Synapses to the stained dendrites derived from bipolar (40.4%) and amacrine (59.6%) cells, whereas outputs from the TH-positive processes were directed to amacrine cells (56.8%) and to small and medium-sized dendrites (35.4%); at least some of these can be considered as ganglion cell dendrites. TH-positive profiles neither formed synapses with each other nor were presynaptic to bipolar cell terminals. Junctional appositions of the immunoreactive profiles were occasionally seen on non-stained amacrine and ganglion cell dendrites in the scleral sublamina of the inner plexiform layer and on optic axons in the optic fibre layer. Although dopaminergic cells are mainly involved in amacrine-amacrine interactions, inputs from bipolar terminals and outputs to ganglion cell dendrites were also substantial, suggestive of a role also in vertical information processing.
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    Identification of different types of serotonin-like immunoreactive olfactory interneurons in four infraorders of decapod crustaceans (1991)
    Springer
    Cell & tissue research 264 (1991), S. 357-362 
    Details
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Olfactory interneurons ; Macrobrachium rosenbergii, Munida sarsi, Pacifastacus leniusculus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum raised against serotonin (5HT) was applied to the brains of representatives of four different infraorders of decapod crustaceans, and revealed two morphological classes of olfactory interneurons. They were classified by the position and size of their cell bodies, and by their connection pattern. One class consisted of giant olfactory interneurons and the other of globuli cells. They were regarded as input and intrinsic interneurons, respectively, because of their morphology. The two classes displayed a similar pattern in two of the infraorders, whereas only one class appeared in the other two infraorders.
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    URL: http://dx.doi.org/10.1007/BF00313974
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    Immunocytochemical localization of muscarinic acetylcholine receptors in the rat endocrine pancreas (1992)
    Springer
    Cell & tissue research 269 (1992), S. 99-106 
    Details
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Immunocytochemistry ; Muscarinic acetylcholine receptor ; Acetylcholine ; Somatostatin ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical application of the antimuscarinic acetylcholine receptor antibody M35 to pancreas tissue revealed the target areas for the parasympathetic nervous system. Immunoreactivity in the endocrine pancreas was much higher than that in the exocrine part. Moreover, the endocrine cells at the periphery of the islets of Langerhans displayed the highest level of immunoreactivity. Based on these findings in the mantle of the islets, two types of islets have been distinguished: type-I islets with intensely stained mantle cells, and type-II islets with a much lower concentration of these cells. On average, type-I islets were larger (244.8 μm±6.1 SEM) than type-II islets (121.5 μm±3.8 SEM). M35-immunoreactivity was present on the majority of D cells, which were characterized by their immunoreactivity to somatostatin [of 446 D cells 356 (79.8%) were M35-immunopositive]. However, only a small proportion of the intensely stained mantle cells belonged to the D cell population. Therefore, it is concluded that the majority of the intensely stained mantle cells represent glucagon-secreting A and/or pancreatic polypeptide-secreting F cells. The intensity of M35-immunoreactivity at the periphery and central core of the islets paralleled the density of cholinergic innervation, suggesting a positive correlation between the intensity of cholinergic transmission and the number of muscarinic acetylcholine receptors at the target structures. The present study further revealed some striking parallels for the muscarinic acetylcholine receptor characteristics between the (endocrine) pancreas and the central nervous system.
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    Immunocytochemical localization of Bombyx-PTTH-like molecules in neurosecretory cells of the brain of the migratory locust, Locusta migratoria (1992)
    Springer
    Cell & tissue research 269 (1992), S. 133-140 
    Details
    ISSN: 1432-0878
    Keywords: Neurosecretory cells ; Brain ; Immunocytochemistry ; Prothoracicotropic hormone ; Insulin-related peptide ; Neuroparsins ; Locusta migratoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a monoclonal antibody directed against a synthetic pentadecapeptide corresponding to the N-terminus of the prothoracicotropic hormone (PTTH) of Bombyx mori, we report the presence of immunoreactive molecules in a large number of median neurosecretory cells of the pars intercerebralis of the migratory locust, Locusta migratoria. These cells correspond to the A1 cell type which we show to contain also neuroparsins, a family of predominant neurohormones of the migratory locust. In contrast, PTTH-like molecules are absent from A2 cells of the pars intercerebralis which contain Locusta insulin-related peptide (LIRP). Developmental studies show the presence of PTTH-related substances in neurosecretory cells of Locusta migratoria from late embryogenesis to adult development, including ageing vitellogenic female adults.
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    Localization of calcitonin gene-related peptide and islet amyloid polypeptide in the rat and mouse pancreas (1992)
    Springer
    Cell & tissue research 269 (1992), S. 315-322 
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    ISSN: 1432-0878
    Keywords: Calcitonin gene-related peptide ; Islet amyloid polypeptide ; Immunocytochemistry ; Pancreas endocrine, exocrine ; Rat (Sprague-Dawley) ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary It was previously demonstrated that the two chemically related peptides calcitonin gene-related peptide (CGRP) and islet amyloid polypeptide (IAPP) both occur in the pancreas. We have now examined the cellular localization of CGRP and IAPP in the rat and the mouse pancreas. We found, in both the rat and the mouse pancreas, CGRP-immunoreactive nerve fibers throughout the parenchyma, including the islets, with particular association with blood vessels. CGRP-immunoreactive nerve fibers were regularly seen within the islets. In contrast, no IAPP-immunoreactive nerve fibers were demonstrated in this location. Furthermore, in rat islets, CGRP immunoreactivity was demonstrated in peripherally located cells, constituting a major subpopulation of the somatostatin cells. Such cells were lacking in the mouse islets. IAPP-like immunoreactivity was demonstrated in rat and mouse islet insulin cells, and, in the rat, also in a few non-insulin cells in the islet periphery. These cells seemed to be identical with somatostatin/CGRP-immunoreactive elements. In summary, the study shows (1) that CGRP, but not IAPP, is a pancreati neuropeptide both in the mouse and the rat; (2) that a subpopulation of rat somatostatin cells contain CGRP; (3) that mouse islet endocrine cells do not contain CGRP; (4) that insulin cells in both the rat and the mouse contain IAPP; and (5) that in the rat, a non-insulin cell population apparently composed of somatostatin cells stores immunoreactive IAPP. We conclude that CGRP is a pancreatic neuropeptide and IAPP is an islet endocrine peptide in both the rat and the mouse, whereas CGRP is an islet endocrine peptide in the rat.
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    5′-Nucleotidase in PC12 cells as revealed by immunocytochemistry (1995)
    Springer
    Cell & tissue research 280 (1995), S. 123-131 
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    ISSN: 1432-0878
    Keywords: Differentiation ; 5′-Nucleotidase ; Immunocytochemistry ; PC12 cells ; Synaptophysin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract 5′-Nucleotidase hydrolyzes 5′-mononucleotides to their nucleosides but is also thought to have a function in neuronal differentiation and synapse formation. The distribution of the enzyme, a glycosyl-phosphatidylinositol-anchored sialoglycoprotein, was investigated in PC12 cells using immunofluorescence microscopy. 5′-Nucleotidase was located both in intracellular compartments and at the cell surface. There was no principal difference in the cellular distribution between undifferentiated cells and after neuritogenic differentiation by nerve growth factor. Intracellularly, 5′-nucleotidase often revealed a sickle-shaped perinuclear distribution and a dotted pattern throughout the cytoplasm, including that of neurites and growth cones. The intracellular distribution was clearly different from that of the synaptic vesicle protein synaptophysin. However, the dotted fluorescence resembled that obtained after uptake of the endosomal marker acridine orange. 5′-Nucleotidase was present on the entire cell surface including all neurites formed after differentiation. There was no increase in 5′-nucleotidase fluorescence at synapse-like contacts between the tips of neurites and other PC12 cells. Surfacelocated 5′-nucleotidase could no longer be detected after the application of glycosyl-phosphatidylinositol-specific phospholipase C to cultured cells. This treatment did not affect PC12 cell differentiation. Our results thus reveal 5′-nucleotidase both at the surface and within organelles and suggest that PC12 cells may be used as a model system for the study of the physiological function of 5′-nucleotidase in neural cells.
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    Origin and destination of globules and irregular masses in the gonadotropin cells from the pituitary of the African catfish, Clarias gariepinus: a morphological study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 113-122 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary ; Gonadotrops ; Crinophagy ; Electron microscopy ; Enzyme cytochemistry ; Immunocytochemistry ; Autoradiography ; Catfish, Clarias gariepinus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The possible function of globules and irregular membrane-bound masses in the gonadotropin cells of the pituitary of Clarias gariepinus was studied. Strong secretory stimulation led to the disappearance of the secretory granules from gonadotropin cells but globules and irregular masses remained present. Acid phosphatase was detected enzyme-cytochemically in both globules and irregular masses. Radiolabelling with tritiated amino acids followed by autoradiography demonstrated that globules received radioactive material after secretory granules. The latter received radioactive material within 75 min of administration of radioactive amino acids but globules and irregular masses did not. Although some globules became radioactively labelled within 24 h of the administration of radioactive amino acids, irregular masses remained unlabelled during this period. Secretory granules reacted positively with antisera against α and β gonadotropin subunits, whereas globules and irregular masses only reacted with the antiserum against the β subunit. A moderate anti-7B2 immunoreactivity was demonstrated in secretory granules and globules, whereas irregular masses labelled strongly. The combined cytological results indicate that globules and irregular masses are degradative, possibly crinophagic structures which develop by fusional events from secretory granules to globules and then to irregular masses.
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    Comparative study of the olfactory epithelium of the three-spined stickleback (Gasterosteus aculeatus) and the nine-spined stickleback (Pungitius pungitius) (1992)
    Springer
    Cell & tissue research 269 (1992), S. 267-273 
    Details
    ISSN: 1432-0878
    Keywords: Olfactory epithelium ; Comparative study ; Histochemistry ; Immunocytochemistry ; Non-specific label ; Microsmatic fish ; Three-spined stickleback, Gasterosteus aculeatus (Teleostei) ; Nine-spined stickleback, Pungitius pungitius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The olfactory epithelium of the three-spined stickleback (Gasterosteus aculeatus) and the nine-spined stickleback (Pungitius pungitius) has been studied with a conventional histochemical and a novel immunological staining technique. In both species, the sensory epithelium is arranged in folds separated by non-sensory epithelial tissue. In the nine-spined stickleback, intrinsic folds consisting of non-sensory cells are found in the apical part of the sensory epithelium where they divide the surface of the sensory epithelium into small islets. These non-sensory cells are non-ciliated, flattened and piled on top of each other; they contain numerous electron-translucent vesicles. The intrinsic folds are absent from the sensory epithelium of the three-spined stickleback. In both species, axons of receptor cells form a layer of fibers in the sensory epithelium immediately above the basal cells. In the three-spined stickleback, thick branches of the olfactory nerve are frequently found in this layer. These branches are only occasionally observed in the sensory epithelium of the nine-spined stickleback. Thus, the three-spined stickleback and the nine-spined stickleback show considerable differences in the organization of the sensory regions of the olfactory epithelium.
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    Development of the diffuse endocrine system in the chicken proventriculus (1993)
    Springer
    Cell & tissue research 271 (1993), S. 107-113 
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    ISSN: 1432-0878
    Keywords: Proventriculus ; Endocrine ontogenesis ; Ultrastructure ; Regulatory peptides ; Immunocytochemistry ; Silver impregnations ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of endocrine cells in the chicken proventriculus has been investigated using light-and electron-microscopy in conjunction with silver and immunocytochemical techniques. The first morphologically detectable endocrine cells were found in 5-day-old embryos by electron microscopy. From the 9th to the 13th day, endocrine cells in contact with the lumen of the organ could be detected both by electron and light (silver impregnation) microscopy. The number of open-type endocrine cells progressively decreased and the number of closed-type increased after this stage. Until the 16th day, endocrine cells were located exclusively in the luminal epithelium, but afterwards they appeared in progressively greater numbers in the compound glands. After hatching, long cytoplasmic processes could be seen in the endocrine cells. Immunoreactivities to regulatory substances appeared in the following order: serotonin (day-14), avian pancreatic polypeptide, glucagon and somatostatin (day-16), bombesin and neurotensin (day-18), and finally, met-enkephalin (day-21).
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    Sexual dimorphism of arg-vasotocin gene expressing neurons in the telencephalon and dorsal diencephalon of the domestic fowl. An immunocytochemical and in situ hybridization study (1996)
    Springer
    Cell & tissue research 287 (1996), S. 69-77 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Vasotocin ; Sex dimorphism ; Bed nucleus of the stria terminalis ; Dehydration ; Immunocytochemistry ; In situ hybridization ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A strong sex dimorphism in the distribution of immunoreactive arginine-vasotocin (AVT) and AVT mRNA was observed in telencephalic and dorsal diencephalic areas of the domestic fowl using immunocytochemistry and in situ hybridization. Two subgroups of immunoreactive parvocellular perikarya surrounded by dense plexus of immunoreactive fibres were found within the bed nucleus of the stria terminalis and the dorsal part of the diencephalic paraventricular region of males. No signs of immunoreactivity were observed within corresponding regions of the female brain. Instead, in females a few scattered weakly stained perikarya were observed rostrally to the level of the anterior commissure, juxtapositioned to the nucleus accumbens and the floor of the lateral ventricle. The distribution of AVT mRNA containing cell profiles fully confirmed the immunocytochemical findings. Osmotic stress induced by water deprivation for 48 h had no influence on the number of immunoreactive or AVT mRNA containing parvocellular cell bodies. However, it resulted in an increase of immunoreactive cell area in the bed nucleus of the stria terminalis and dorsal diencephalon of 5.9 and 11.7%, respectively. We suggest that the sexually dimorphic vasotocinergic circuit may be involved in the co-ordination of behavioural and autonomic functions in response to environmental stress.
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    Morphology and distribution of Müller cells in the retina of the toad Bufo marinus (1993)
    Springer
    Cell & tissue research 272 (1993), S. 183-192 
    Details
    ISSN: 1432-0878
    Keywords: Retina ; Müller cells ; Neuron-specific enolase ; Immunocytochemistry ; Quantitative analysis ; Ultrastructure ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have previously shown that an antibody against neuron-specific enolase (NSE) selectively labels Müller cells (MCs) in the anuran retina (Wilhelm et al. 1992). In the present study the light- and electron-microscopic morphology of MCs and their distribution were described in the retina of the toad, Bufo marinus, using the above antibody. The somata of MCs were located in the proximal part of the inner nuclear layer and were interconnected with each other by their processes. The MCs were uniformly distributed across the retina with an average density of 1500 cells/mm2. Processes of MCs encircled the somata of photoreceptor cells isolating them from each other by glial sheath, except for those of the double cones. Some of the photoreceptor pedicles remained free of glial sheath. Electron-microscopic observations confirmed that MC processes provide an extensive scaffolding across the neural retina. At the outer border of the ganglion cell layer these processes formed a non-continuous sheath. The MC processes traversed through the ganglion cell layer and spread beneath it between the neuronal somata and the underlying optic axons. These processes formed a continuous inner limiting membrane separating the optic fibre layer from the vitreous tissue. Neither astrocytic nor oligodendrocytic elements were found in the optic fibre layer. The significance of the uniform MC distribution and the functional implications of the observed pattern of MC scaffolding are discussed.
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    Immunocytochemical studies of chicken somatotrophs and somatotroph granules before and after hatching (1993)
    Springer
    Cell & tissue research 272 (1993), S. 369-374 
    Details
    ISSN: 1432-0878
    Keywords: Somatotrophs ; Somatotroph granules ; Growth hormone ; Immunocytochemistry ; Developments ontogenic ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemical methods were used to gain information about the embryonic development of chicken somatotrophs before and after hatching. To localize growth hormone, anterior pituitary sections were incubated with growth-hormone antibody, and then an indirect peroxidase method was used for light microscopy and an immunogold method for electron microscopy. The earliest evidence of embryonic somatotrophs was seen at 12 days. At this stage somatotrophs were sparse (0.2% of parenchymal cells) and their granules were pleomorphic with elongated ovoid and lozenge shapes predominating. Few of the immunogold-labeled somatotroph granules of the embryo were spherical until 15 days after fertilization. At 18 days, most of the granules were spherical (their shape in the adult chicken). During the six days between the 15-day-old embryo and the 1-day-old chick, the number of gold particles per granule section approximately doubled suggesting an increase in growth hormone content of the granules. This rise was the result of increases in the size of the granule sections and in the concentration of gold particles in the sections. During the embryonic period of 12–20 days, somatotrophs were not more than 3.6% of the anterior pituitary cell population. During the following two days, between the 20-day-old embryo and the 1-day-old chick, the percentage of somatotrophs in the pituitary parenchymal cell population rose rapidly from 3.6% to 20.7% and then increased slowly to 24.6% during the period of 1–5 days after hatching. Both the sharp percentage rise in somatotrophs (20-day-old embryo to 1-day-old chick) and the rise in growth hormone content of the granules (15-day-old embryo to 1-day-old chick) suggested by gold-particle counts occur close to the time of hatching. These morphological changes may reflect an increased synthesis of growth hormone that is responsible for the rise in plasma growth-hormone concentration that begins about the same time and is especially abrupt two days later (1–3 days after hatching).
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    Irregular distribution of mammary-derived growth inhibitor in the bovine mammary epithelium (1993)
    Springer
    Cell & tissue research 272 (1993), S. 383-389 
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    ISSN: 1432-0878
    Keywords: Mammary gland ; Growth inhibitor ; Epithelium ; Cell types ; Differentiation ; Immunocytochemistry ; Immunohistochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Localization of a mammary-derived growth inhibitor (MDGI) in the bovine mammary gland was verified by light-and electron-microscopic methods. Expression of MDGI, which is known to inhibit the growth of mammary epithelial cell lines in vitro, was found to be highest in the late pregnant and in the lactating state. A combination of immunohistochemical and immunocytochemical methods with semi- and ultrathin resin sections revealed marked variations in MDGI staining. High MDGI levels were predominantly detectable in epithelial cells with large milk fat droplets. Distinct cell types that were almost free of label could be identified among bovine mammary epithelial cells that always exhibited high MDGI levels. Similar results were obtained when using a serum-free organ culture system in which MDGI was hormonally induced in cell types of comparable differentiation state. The specific occurrence of the growth inhibitor in developing alveoli and certain cell types points to the association between MDGI expression and functional differentiation in the normal mammary gland.
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    Ultrastructural distribution of endogenous immunoglobulin-G in human term amniochorion (1995)
    Springer
    Cell & tissue research 281 (1995), S. 367-374 
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    ISSN: 1432-0878
    Keywords: Placenta ; Amniochorion ; Cytotrophoblast cells ; Immunoglobulin G ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Maternal immunoglobulin-G (IgG) is known to be transported across the placental syncytiotrophoblast during the period when the human fetus is incapable of manufacturing these defensive molecules. In this study we investigated the possible role of the amniochorion, that surrounds the amniotic cavity in which the fetus lies, in the transfer of immunoglobulin. Endogenous IgG was localised in the amniochorion by confocal immunofluorescence microscopy and by ultrastructural labelling of ultrathin frozen tissue sections using the protein A-gold technique. Immunoreactivity was identified in the extracellular matrix tissues and necrotic amniotic epithelial cells. Healthy amniotic epithelial cells and cytotrophoblast cells of the chorion laeve were devoid o endogenous IgG. These results suggest a possible non-specific paracellular transport pathway between cytotrophoblast cells, which may conceivably contribute to the acquisition of passive immunity by the fetus, and offer a rational explanation for the presence of small quantities of maternal IgG in the amniotic fluid.
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    Distribution of synapses on two local auditory interneurones, ON1 and ON2, in the prothoracic ganglion of the cricket: relationships with GABA-immunoreactive neurones (1996)
    Springer
    Cell & tissue research 283 (1996), S. 231-246 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Auditory system ; Nervous system ; insect ; Interneurons ; Immunocytochemistry ; Inhibitory synapses ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In the prothoracic ganglia of the cricket Gryllus bimaculatus two local auditory interneurones, ON1 and ON2, were labelled for electron microscopy by intracellular injection of horseradish peroxidase following physiological characterisation. The neurones branch in the median ventral association centre and the root of nerve 5 on both sides of the ganglion. As they are very similar in shape and position they may share a common embryological origin. Differences are found in the details of the fine branching pattern and in their physiology as ON1 is tuned particularly to low sound frequencies of 4–5 kHz whereas ON2 is more sensitive to frequencies above 8 kHz. Although the ON1 neurones inhibit each other and are involved in the inhibition of other auditory neurones they were not labelled by antibodies against the inhibitory transmitter GABA and their vesicles differ significantly from those in neurones that are. The same is true of the ON2 neurones whose vesicles also differ significantly from those in ON1 supporting light-microscope evidence that they may use different transmitters. The distribution of input and output synapses on the ipsilateral and contralateral branches of ON1 and ON2, and the proportion of the synapses made from and onto neuropilar processes immunoreactive for GABA was determined. In ON1 94% of the input synapses were received on the ipsilateral branches and 62% of the outputs made from the contralateral branches. This confirms previous physiological evidence that input is received ipsilaterally and output made contralaterally but the presence of some contralateral input and a significant ipsilateral output was unsuspected. Thirty percent of the input synapses on the ipsilateral side and 75% on the contralateral side were made from GABA-immunoreactive processes but processes postsynaptic to ON1 were rarely immunoreactive. The distribution of input synapses on ON2 was similar with 90% received on ipsilateral branches but a higher proportion of outputs (83%) was made from the contralateral side than in ON1. Thirty one percent of ipsilateral inputs were GABA-immunoreactive but only 14% on the contralateral side.
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    Neuropeptide Y in the forebrain and retina of the killifish, Fundulus heteroclitus (1996)
    Springer
    Cell & tissue research 283 (1996), S. 313-323 
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    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Brain ; Pituitary ; Pineal organ ; Retina ; CSF-contacting neurons ; Immunocytochemistry ; Killifish ; Fundulus heteroclitus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The organization of the neuropeptide Y (NPY)-immunoreactive system in the forebrain, pineal organ and retina of a biweekly spawning fish (Fundulus heteroclitus) was investigated. Immunoreactivity was encountered in neurons of the nucleus olfactoretinalis, in the large population of neurons in the floor of the telencephalon, and in the nucleus entopeduncularis. Isolated somata were encountered in the hypophysiotropic hypothalamic nuclei, viz., the nucleus preopticus periventricularis, nucleus preopticus, and nucleus lateralis tuberis. Immunoreactive somata were also seen in the nucleus dorsomedialis thalami. The olfactory bulb was abundantly innervated by NPY fibers. The telencephalon showed thick radiating processes basally and terminal fields with modest to high densities in the dorsal and lateral regions. NPY-immunoreactive fibers were also conspicuous in the preoptic area, suprachiasmatic nucleus, tuberal hypothalamus, pituitary gland, and paraventricular thalamic regions; discrete CSF-contacting sites were also encountered. Of special interest was the occurrence of NPY immunoreactivity in fibers of the pineal stalk and organ. In the retina, some amacrine cells displayed immunoreactivity, while the inner plexiform layer revealed a well-developed pattern of NPY fibers different from that previously described for the goldfish.
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    Immunolocalization of Na+, K+-ATPase in the gill epithelium of rainbow trout, Oncorhynchus mykiss (1996)
    Springer
    Cell & tissue research 283 (1996), S. 461-468 
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    ISSN: 1432-0878
    Keywords: Key words: Gills ; Na+ ; K+-ATPase ; Immunocytochemistry ; DASPMI ; Cell culture ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The Na+,K+-ATPase (the sodium pump) plays a crucial role in ion transport in the fish gill. An immunocytochemical method has been optimized, using the mouse monoclonal antibody IgG α5, raised against the αsubunit of the avian sodium pump, to localize Na+,K+-ATPase in fish gill cells. The method appears to be successful for the immunolocalization of Na+,K+-ATPase in both paraffin-embedded gill tissue sections and primary cultures of fish gill epithelial cells. The immunostaining has demonstrated that Na+,K+-ATPase-positive cells are mainly localized on the primary lamellae, in the interlamellar region, which is in agreement with the distribution of ion-transporting cells, also called chloride cells, as shown by electron microscopy. Na+,K+-ATPase-positive cells have been demonstrated for the first time in primary cultures of gill epithelial cells. Comparative labeling studies of primary cultures have shown that sites of Na+,K+-ATPase-positive cells correspond to sites of cells labeled with dimethylaminostyrylmethyl-pyridiniumiodine, a fluorescent mitochondrial probe for ion-transporting cells. The immunocytochemical detection method for Na+,K+-ATPase in cells is proposed as an easy and specific Na+-transport-related method to characterize and localize ion-transporting cells in primary cultures and in tissue sections of fish gill epithelium.
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    Localization of corticotropin-releasing factor immunoreactivity in the brain of the teleost Sparus aurata (1995)
    Springer
    Cell & tissue research 281 (1995), S. 569-572 
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    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor ; Immunocytochemistry ; Gilthead sea bream ; Sparus aurata (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of perikarya and fibers containing corticotropin-releasing factor (CRF) was studied in the brain of the teleost Sparus aurata by immunocytochemistry using the peroxidase-antiperoxidase method. Antisera against rat CRF, arginine vasotocin, and human adrenocorticotropin (ACTH) were used. Most CRF-immunoreactive neurons were located in the nucleus lateralis tuberis, but they were absent from the nucleus preopticus, which only contained arginine vasotocin neurons. Few CRF perikarya were identified in the nucleus preopticus periventricularis and in the mesencephalic tegmentum. A conspicuous bundle of immunoreactive fibers ran along the diencephalic floor and pituitary stalk to end near the cells of the hypophysial pars intermedia. No CRF was seen near the adenohypophysial rostral pars distalis. Our results suggest that, in Sparus aurata, CRF is a releasing factor for melanotropic cells. Its role as a releasing factor for ACTH is discussed.
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    Occurrence of enzymes of free radical metabolism suggests the possible cytotoxic capacity of the transitional epithelium of the human ureter (1997)
    Springer
    Cell & tissue research 287 (1997), S. 351-356 
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    ISSN: 1432-0878
    Keywords: Key words: Urothelium ; Tartrate-resistant acid phosphatase ; Nitric oxide synthase I ; Superoxide dismutase ; Immunocytochemistry ; Free radicals ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three enzymes, viz., tartrate-resistant acid phosphatase (TRAP), nitric oxide synthase I (NOS-I), and superoxide dismutase (SOD), involved in the production and metabolism of free radicals or radical equivalents, were demonstrated by immunocytochemistry in the urothelium of the ureters of six patients of various ages. Two of these enzymes (TRAP and NOS-I) were colocalized in the most apical and lateral border of the superficial cells of the urothelium. In contrast, SOD showed a patchy or granular distribution within the supranuclear region of these cells. Intra- and subepithelial macrophages exhibited a weak TRAP, but no NOS-I or SOD, immune reaction. On the basis of the immunocytochemical findings, arguments in favor of a cytotoxic function of the superficial cells of the human urothelium are presented.
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    Presence of embryonic myosin in normal postural muscles of the adult rat (1995)
    Springer
    Cell & tissue research 280 (1995), S. 541-548 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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    Sources of inputs to longitudinal muscle motor neurons and ascending interneurons in the guinea-pig small intestine (1995)
    Springer
    Cell & tissue research 280 (1995), S. 549-560 
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    ISSN: 1432-0878
    Keywords: Key words: Enteric nervous system ; Immunocytochemistry ; Calretinin ; Calbindin ; Bombesin ; Small intestine ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guinea-pig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin- and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.
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    Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves associated with coronary blood vessels of rat and guinea-pig (1995)
    Springer
    Cell & tissue research 280 (1995), S. 575-582 
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    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase ; Coronary vasculature ; Electron microscopy ; Immunocytochemistry ; Rat (Sprague Dawley) ; Guinea-pig (Dunkin Hartley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ultrastructural investigation of nitric oxide synthase-immunoreactive nerves closely associated with blood vessels in rat and guinea-pig hearts revealed many labelled nerve fibres in the walls of the main branches of the coronary arteries, and in arterioles, capillaries and post-capillary venules. The number of nitric oxide synthase-containing nerve fibres associated with different vessels, even those of the same calibre, varied. Terminal regions of nitric oxide synthase-immunoreactive fibres were observed in the endocardium and myocardium. Nitric oxide synthase-labelled fibres displayed electron-dense immunoproduct in both varicose and intervaricose regions. Immunoreactive axonal varicosities contained both small and large synaptic vesicles. The characteristics of the nitric oxide synthase-immunoreactive nerve fibres observed in the heart and the possibility that these fibres represent the processes of intracardiac neurones and/or sensory neurones of extrinsic origin are discussed.
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    The distribution of neurones immunoreactive for β-tyrosine hydroxylase, dopamine and serotonin in the ventral nerve cord of the cricket, Gryllus bimaculatus (1995)
    Springer
    Cell & tissue research 280 (1995), S. 583-604 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Dopamine ; Serotonin ; Tyrosine hydroxylase ; Immunocytochemistry ; Nervous system ; insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine- (n≤70) and serotonin-immunoreactive (n≤120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine- and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.
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    Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium (1995)
    Springer
    Cell & tissue research 281 (1995), S. 77-83 
    Details
    ISSN: 1432-0878
    Keywords: Esophagus ; Epithelial cells ; Intestinal lectin, L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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    Specific immunocytochemical localization of cathepsin E at the ruffled border membrane of active osteoclasts (1995)
    Springer
    Cell & tissue research 281 (1995), S. 85-91 
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    ISSN: 1432-0878
    Keywords: Cathepsin E ; Aspartic proteinase ; Osteoclasts ; Immunocytochemistry ; Rat (WKA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunocytochemical localization of cathepsin E, a non-lysosomal aspartic proteinase, was investigated in rat osteoclasts using the monospecific antibody to this protein. At the light-microscopic level, the preferential immunoreactivity for cathepsin E was found at high levels in active osteoclasts in the physiological bone modeling process. Neighboring osteoblastic cells were devoid of its immunoreactivity. At the electron-microscopic level, cathepsin E was exclusively confined to the apical plasma membrane at the ruffled border of active osteoclasts and the eroded bone surface. Cathepsin E was also concentrated in some endocytotic vacuoles of various sizes in the vicinity of the ruffled border membrane, some of which appeared to be secondary lysosomes containing the phagocytosed materials. These results strongly suggest that this enzyme is involved both in the extracellular degradation of the bone organic matrix and in the intracellular breakdown of the ingested substances in osteoclasts.
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    Localization of integrin subunits α6 and β1 during somitogenesis in the long-tailed macaque (M. fascicularis) (1995)
    Springer
    Cell & tissue research 281 (1995), S. 101-108 
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    ISSN: 1432-0878
    Keywords: Somite ; Intergin ; Extracellular matrix, structures ; Embryo ; Laminin ; Immunocytochemistry ; Macaca fascicularis (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of integrin subunits α6 and β1, and the α6β1 integrin ligand, laminin, was examined during somitogenesis in developmental stages 11, 13, and 16 in the long-tailed macaque, using peroxidase immunocytochemistry. Within differentiating somites in stage 11, α6 expression was observed in the sclerotome, basal surface of dermamyotomal cells adjacent to the basal lamina and on scattered cells throughout the dermamyotome. In further advanced somites in stages 13 and 16, α6 immunoreactivity become restricted to the myotome, α6 was expressed on mesenchymal core cells within the myocele of undifferentiated epitheliod somites and the ventromedial wall of somites commencing differentiation at each stage. β1 distribution resembled that of α6 in stage 11 somitic tissue, however, it remained present on myotome and sclerotome cells in the later stages, and was also expressed on dermatomal cells in stage 16. Laminin immunoreactivity, while more intense and prevalent than α6 and β1 in each stage examined, occurred on the same somite cell populations as the 2 integrin subunits. These results show a defined distribution of α6 on somitic tissue, and suggest this integrin is involved in somite differentiation. They also support a possible role for α6 in myoblast formation and migration. Overlapping of β1 and laminin immunoreactivity with that of α6 further suggests that α6 paris with β1 as a functional heterodimer for laminin in defined somitic regions.
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    Immunocytochemical, electrophoresis, and immunoblot analysis of Heliothis virescens gap junctions isolated in the presence and absence of protease inhibitors (1995)
    Springer
    Cell & tissue research 281 (1995), S. 179-186 
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    ISSN: 1432-0878
    Keywords: Gap junction ; Intercellular junction ; Insect ; Arthropod ; Immunocytochemistry ; Hellothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the ∼10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an ∼40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an ∼30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.
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    Immunocytochemical analysis of the transport of arginine analogues into nitrergic neurons and other cells in the retina and pituitary (1997)
    Springer
    Cell & tissue research 290 (1997), S. 501-514 
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    ISSN: 1432-0878
    Keywords: Key words: Arginine ; Analogues ; Immunocytochemistry ; Nitric oxide ; Pituitary ; Retina ; Transport ; Rat (Wistar) ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Nitric oxide is formed by the action of nitric oxide synthase upon l-arginine. The efficacy of some exogenously applied arginine analogues in inhibiting nitric oxide synthase and thus nitrergic transmission indicates that neurons producing nitric oxide may possess an arginine transport system. To investigate whether arginine analogues are preferentially transported into nitric oxide-utilising cells or into cells making other neurochemicals, we have raised highly specific antisera against a number of arginine analogues including NG-methyl arginine, d-arginine, NGnitro-l-arginine, NG-nitro-l-arginine methyl ester and canavanine. Retinae were incubated in physiological media containing these analogues and rats were given intraperitoneal injections of the analogues to study the pituitary. Immunocytochemistry and NADPH-diaphorase histochemistry revealed that many of these analogues could be transported preferentially, but not exclusively, into nitric oxide-generating cells. However, some nitric oxide-producing cells apparently lacked the ability to take up some arginine analogues. We conclude that nitric oxide-generating cells in the retina and pituitary possess one or more arginine transporters. Other subsets of neurons that use GABA or glutamate as a neurotransmitter may also accumulate arginine analogues, possibly as a substrate for formation of these neurochemicals.
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    URL: http://dx.doi.org/10.1007/s004410050957
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    A comparative immunocytochemical study using an antiserum against a synthetic analogue of the corpora cardiaca peptide Pea-CAH-I (MI, neurohormone D) of Periplaneta americana (1996)
    Springer
    Cell & tissue research 284 (1996), S. 401-413 
    Details
    ISSN: 1432-0878
    Keywords: Key words: AKH/RPCH peptide family ; Insect brain ; Corpora cardiaca ; Immunocytochemistry ; Enzyme immunoassay ; Periplaneta americana ; Leucophaea maderae (Insecta) ; Tegenaria atrica (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. An antiserum against the octapeptide Pea-CAH-I, a member of the adipokinetic hormone/red pigment-concentrating hormone family, has been produced for immunocytochemical staining in insects and various other invertebrate species. The anti-Pea-CAH-I serum stains the glandular corpora cardiaca cells of those insect species that synthesize identical or structurally similar peptides. In the corpora cardiaca of species producing peptides with a different C-terminus, these cells remain unstained. Pea-CAH-I-like immunoreactivity has also been found in neurons of the central nervous system of all invertebrate orders studied. The antiserum recognizes the C-terminal sequence Pro-Asn-Trp-NH2 of the Pea-CAH-I molecule as established by enzyme immunoassay. The widespread Pea-CAH-I-like immunoreactivity in all nervous systems of the studied animals probably does not reflect the presence of Pea-CAH-I but the occurrence of peptides carrying similar epitopes.
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    URL: http://dx.doi.org/10.1007/s004410050601
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    Immunocytochemical characterization of the accessory medulla in the cockroach Leucophaea maderae (1995)
    Springer
    Cell & tissue research 282 (1995), S. 3-19 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS) ; invertebrate ; Optic lobe ; Pigment-dispersing hormone ; insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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    Ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the dogfish (1995)
    Springer
    Cell & tissue research 282 (1995), S. 33-40 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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    Caveolin and its cellular and subcellular immunolocalisation in lung alveolar epithelium: implications for alveolar epithelial type I cell function (1999)
    Springer
    Cell & tissue research 295 (1999), S. 111-120 
    Details
    ISSN: 1432-0878
    Keywords: Key words Caveolin ; Caveolae ; Lung ; Alveolar epithelial type I cell ; Immunocytochemistry ; Electron microscopy ; Confocal laser scanning microscopy ; Rat (CD)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Caveolae are flask-shaped invaginations of the plasmalemma which pinch off to form discrete vesicles within the cell cytoplasm. Biochemically, caveolae may be distinguished by the presence of a protein, caveolin, that is the principal component of filaments constituting their striated cytoplasmic coat. Squamous alveolar epithelial type I (ATI) cells, comprising approximately 95% of the surface area of lung alveolar epithelium, possess numerous plasmalemmal invaginations and cytoplasmic vesicles ultrastructurally indicative of caveolae. However, an ultrastructural appearance does not universally imply the biochemical presence of caveolin. This immunocytochemical study has utilised a novel application of confocal laser scanning and electron microscopy unequivocally to localise caveolin-1 to ATI cells. Further, cytoplasmic vesicles and flask-shaped membrane invaginations in the ATI cell were morphologically identified whose membranes were decorated with anti-caveolin-1 immunogold label. Coexistent with this, however, in both ATI and capillary endothelial cells could be seen membrane invaginations morphologically characteristic of caveolae, but which lacked associated caveolin immunogold label. This could reflect a true biochemical heterogeneity in populations of morphologically similar plasmalemmal invaginations or an antigen threshold requirement for labelling. The cuboidal alveolar epithelial type II cell (ATII) also displayed specific label for caveolin-1 but with no ultrastructural evidence for the formation of caveolae. The biochemical association of caveolin with ATI cell vesicles has broad implications for the assignment and further study of ATI cell function.
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    URL: http://dx.doi.org/10.1007/s004410051217
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    Periviscerokinin-like immunoreactivity in the nervous system of the American cockroach (1999)
    Springer
    Cell & tissue research 295 (1999), S. 159-170 
    Details
    ISSN: 1432-0878
    Keywords: Key words Neuropeptides ; Perisympathetic organ ; Myotropin ; Visceral muscles ; Immunocytochemistry ; Insect nervous system ; Periplaneta americana (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A highly specific polyclonal antiserum has been raised against periviscerokinin, the first neuropeptide isolated from the perisympathetic organs of insects (Predel et al. 1995). In this study, two different neuronal systems with periviscerokinin-like immunoreactivity were distinguished in the central nervous system of the American cockroach: (1) An intrinsic neuronal network, restricted to the head-thoracic region, was formed by intersegmental projecting neurons of the brain, suboesophageal ganglion and metathoracic ganglion. In addition, groups of local interneurons occurred in the proto- and tritocerebrum. (2) A typical neurohormonal system was stained exclusively in the abdomen; it was represented by abdominal perisympathetic organs which were supplied by three cell clusters located in each unfused abdominal ganglion. As revealed by nickel backfills, most neurons with axons entering the perisympathetic organs contained a periviscerokinin-like peptide. Immunoreactive fibres left the perisympathetic organs peripherally, innervated the hyperneural muscle and ran via the link nerves/segmental nerves to the heart and segmental vessels. All visceral muscles innervated by periviscerokinin-immunoreactive fibres were shown to be sensitive to periviscerokinin, whereas the hindgut gave no specific response to this peptide.
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    Localization of choline acetyltransferase-expresing neurons in the larval vissual system of Drosophila melanogaster (1995)
    Springer
    Cell & tissue research 282 (1995), S. 193-202 
    Details
    ISSN: 1432-0878
    Keywords: Choline acetyltransferase ; Cholinergic neuron ; Visual system ; Bolwig's organ ; Immunocytochemistry ; In situ hybridization ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Choline acetyltransferease (ChAT) is the enzyme catalyzing the biosynthesis of acetylcholine and is considered to be a phenotypically specific marker for cholinergic neurons. We have examined the distribution of ChAT-expressing neurons in the larval nervous system of Drosophila melanogaster by three different but complementary techniques: in situ hybridization with a cRNA probe to ChAT messenger RNA, immunocytochemistry using a monoclonal anti-ChAT antibody, and X-gal staining of transformed animals carrying a reporter gene composed of 7.4 kb of 5′ flanking DNA from the ChAT gene fused to a lacZ reporter gene. All three techniques demonstrated ChAT-expressing neurons in the larval visual system. In embryos, the photoreceptor organ (Bolwig's organ) exhibited strong cRNA hybridization signals. The optic lobe of late third-instar larvae displayed ChAT immunoreactivity in Bolwig's nerve and a neuron close to the insertion site of the optic stalk. This neuron's axon ran in parallel with Bolwig's nerve to the larval optic neuropil. This neuron is likely to be a first-order interneuron of the larval visual system. Expression of the lacZ reporter gene was also detected in Bolwig's organ and the neuron stained by anti-ChAT antibody. Our observations indicate that acetylcholine may be a neurotransmitter in the larval photoreceptor cells as well as in a first-order interneuron in the larval visual system of Drosophila melanogaster.
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    Ultrastructural, morphometrical and immunocytochemical analyses of the exocrine pancreas in a hibernating dormouse (1998)
    Springer
    Cell & tissue research 292 (1998), S. 531-541 
    Details
    ISSN: 1432-0878
    Keywords: Key words Pancreas ; exocrine ; Hibernation ; Amylase ; Immunocytochemistry ; Muscardinus avellanarius (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Pancreatic acinar cells of euthermic, hibernating and arousing individuals of the hazel dormouse Muscardinus avellanarius (Gliridae) have been observed at the electron-microscopic level and analysed by means of ultrastructural morphometry and immunocytochemistry in order to investigate possible fine structural changes of cellular components during periods of strikingly different degrees of metabolic activity. During hibernation, the cisternae of the rough endoplasmic reticulum (RER) flatten assuming a parallel pattern, the Golgi apparatus is extremely reduced and the mitochondria contain many electron-dense particles. The cell nuclei appear irregularly shaped, with deep indentations containing small zymogen granules. They also contain abundant coiled bodies and unusual constituents, such as amorphous bodies and dense granular bodies. Large numbers of zymogen granules occur in all animals. However, the acinar lumina are open and filled with zymogen only in euthermic animals, whereas, in hibernating and arousing individuals, they appear to be closed. Morphometrical analyses indicate that, in pancreatic acinar cells, nuclei and zymogen granules significantly decrease in size from euthermia to hibernation, probably reflecting a drastic decrease of metabolic activities, mainly protein synthesis and processing. In all the studied animals, immunocytochemistry with specific antibodies has revealed an increasing gradient in α-amylase content along the RER-Golgi-zymogen granule pathway, reflecting the protein concentration along the secretory pathway. Moreover, during deep hibernation, significantly larger amounts of α-amylase accumulate in RER and zymogen granules in comparison to the other seasonal phases analysed. Upon arousal, all cytoplasmic and nuclear constituents restore their euthermic aspect and all morphometrical and immunocytochemical parameters exhibit the euthermic values, thereby indicating a rapid resumption of metabolic activities.
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    Expression pattern for adrenomedullin during pancreatic development in the rat reveals a common precursor with other endocrine cell types (1998)
    Springer
    Cell & tissue research 293 (1998), S. 95-100 
    Details
    ISSN: 1432-0878
    Keywords: Key words Adrenomedullin ; Pancreas ; Development ; Immunocytochemistry ; Colocalizations ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Adrenomedullin is an α-amidated 52-amino acid peptide involved in many physiological actions, among others the regulation of insulin secretion. Using immunohistochemical methods, we found that adrenomedullin immunoreactivity first appears at day 11.5 of embryonic development in the rat, coinciding with the appearance of pancreatic glucagon. The early appearance of adrenomedullin in the developing pancreas may indicate an active involvement in either the morphogenesis of the organ or its endocrine/paracrine/autocrine hormone regulation during intrauterine life. We also investigated the pattern of colocalizations of adrenomedullin with the other pancreatic hormones. At some point during development all the cell types express adrenomedullin, progressively evolving towards the adult pattern where only the pancreatic polypeptide cells contain a strong immunoreactivity for adrenomedullin. At this point the remaining cells of the islet are, in general, weakly stained. This sequential and time-dependent expression of adrenomedullin suggests a tight regulation similar to that observed for other modulatory substances responsible for embryonic morphogenesis.
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    Immunocytochemistry and in situ hybridization studies of pepsinogen C-producing cells in developing rat fundic glands (1998)
    Springer
    Cell & tissue research 293 (1998), S. 121-131 
    Details
    ISSN: 1432-0878
    Keywords: Key words Pepsinogen C ; Ontogeny ; Mucous neck cell ; Chief cell ; Intermediate mucopeptic cell ; Immunocytochemistry ; In situ hybridization ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The ontogeny of pepsinogen C-producing cells in rat fundic glands was studied by means of light and electron microscopy using an antiserum raised against a synthetic peptide based on rat pepsinogen C. To confirm the immunocytochemistry results, the expression of rat pepsinogen C messenger RNA (mRNA) in the fundic gland was also examined by in situ hybridization using a digoxigenin-labeled RNA probe. In adult rats, pepsinogen C was produced by chief cells, mucous neck cells, and intermediate mucopeptic cells. Pepsinogen C-producing cells appeared in embryos as early as 18.5 days’ gestation. The development of these cells could be classified into four stages: (1) 18.5 days’ gestation to 0.5 days after birth; (2) 0.5 days to 2 weeks after birth; (3) 3–4 weeks after birth; (4) 4–8 weeks after birth. In embryos and young animals, pepsinogen C-producing cells were mucopeptic cells. By 4 weeks after birth, mucous neck cells could be distinguished morphologically. The maturation stages of the chief cells could be traced by electron microscopy along the longitudinal axis of the rat fundic gland by double-staining with anti-pepsinogen C antibody and periodic acid-thiocarbohydrazide-silver proteinate. Positive reactions for pepsinogen C and pepsinogen C mRNA expression were detected in mucous neck cells. Therefore, we conclude that mucous neck cells are precursor cells of chief cells. Mucous neck cells, intermediate cells, and chief cells are in the same differentiating cell lineage.
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    Quantitative analysis of inputs to somatostatin-immunoreactive descending interneurons in the myenteric plexus of the guinea-pig small intestine (1998)
    Springer
    Cell & tissue research 294 (1998), S. 219-226 
    Details
    ISSN: 1432-0878
    Keywords: Key words Enteric nervous system ; Somatostatin ; Immunocytochemistry ; Intestinal motility ; Synaptic connections ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Somatostatin immunoreactivity occurs in a specific subgroup of cholinergic descending interneurons in the myenteric plexus of the guinea-pig small intestine. In the present work, we made light- and electron-microscopic investigations of chemically defined inputs to these neurons, in order that the origins of the connections of other neurons with them could be deduced. Somatostatin-immunoreactive synapses and close contacts were found on the cell bodies and filamentous processes of somatostatin neurons; these were 84% of all inputs. It is thus confirmed that this class of interneuron forms chains that project anally. Descending interneurons with immunoreactivity for nitric oxide synthase provided 14% of inputs to somatostatin-immunoreactive descending interneurons. An antiserum against a calcium-binding protein, calbindin, was used as marker for the majority of intrinsic primary afferent neurons, AH/Dogiel type II neurons; this class of neurons provided only 2.5% of the inputs to somatostatin-immunoreactive descending interneurons. We conclude that somatostatin-immunoreactive descending interneurons are involved in the conduction of impulses distally along the full length of the small intestine, but receive only a minor input from calbindin-immunoreactive primary afferent neurons.
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    Morphology and synaptic connectivity of nitric oxide synthase-immunoreactive neurons in the guinea pig retina (1999)
    Springer
    Cell & tissue research 297 (1999), S. 397-408 
    Details
    ISSN: 1432-0878
    Keywords: Key words Retina ; NOS ; Immunocytochemistry ; Synaptic connectivity ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Immunocytochemical methods with an antiserum against neuronal nitric oxide synthase (NOS) were applied to identify the morphology and synaptic connectivity of NOS-like immunoreactive neurons in the guinea pig retina. In the present study, two types of amacrine cells were labeled with anti-NOS antisera. Type 1 cells had large somata located in the inner nuclear layer (INL) with long, sparsely branched processes ramifying mainly in stratum 3 of the inner plexiform layer (IPL). The somata of type 2 cells (smaller diameters) were located in the INL. Some displaced amacrine cells in the ganglion cell layer were labeled. The soma size of the displaced amacrine cells was similar to that of the type 2 amacrine cells. However, processes originating from type 2 amacrine cells and displaced amacrine cells stratified mainly in strata 1 and 5, respectively. Some cone bipolar cells were weakly NOS-immunoreactive. The synaptic connectivity of NOS-like immunoreactive amacrine cells was identified in the IPL by electron microscopy. NOS-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in all strata of the IPL. The most frequent postsynaptic targets of NOS-immunoreactive amacrine cells were other amacrine cell processes. Cone bipolar cells were postsynaptic to NOS-labeled amacrine cells in all strata of the IPL. Labeled amacrine cells synapsing onto ganglion cells were found only in sublamina b. A few synaptic contacts were observed between labeled cell processes. In the outer plexiform layer, dendrites of labeled bipolar cells made basal contact with cone pedicles or formed a synaptic triad opposed to a synaptic ribbon of cone pedicles.
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    Pineal-retinal relationships: rhythmic biosynthesis and immunocytochemical localization of melatonin in the retina of the pike (Esox lucius) (1991)
    Springer
    Cell & tissue research 265 (1991), S. 601-609 
    Details
    ISSN: 1432-0878
    Keywords: Retina ; Melatonin ; Rhythmic biosynthesis ; Immunocytochemistry ; Pike, Esox lucius (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The levels of melatonin and the activities of two enzymes of the melatonin biosynthetic pathway, serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), were measured throughout the light-dark cycle in the retina of a teleost fish, the pike. HIOMT activity did not display significant variations, whereas NAT activity and melatonin content showed a daily rhythm, high levels occurring during the night. The profiles of the latter two rhythms did not closely match one another and differed from those previously described in the pineal organ of the same species. These results are discussed with respect to a possible paracrine role of retinal melatonin. Melatonin-like immunoreactivity was found in the photoreceptor cell layer and in the Müller cells of the inner nuclear layer. The intensity of the melatonin-like immunoreactivity varied throughout the 24 h light-dark cycle, in good correlation with the variations in the melatonin level as measured by radioimmunoassay.
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    Stereological study of gonadotropes in the frog, Rana pipiens, after GnRH stimulation in vitro (1990)
    Springer
    Cell & tissue research 262 (1990), S. 171-176 
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    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Gonadotropes ; Morphometry ; Stereology ; Rana pipiens (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or “storage” pool of gonadotropin is associated mainly with the small and medium secretory granules.
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    Immunohistochemical distribution of simple-epithelial-type keratins and other intermediate filament proteins in the developing human pituitary gland (1991)
    Springer
    Cell & tissue research 266 (1991), S. 59-64 
    Details
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Rathke's pouch ; Intermediate filaments ; Cytokeratins ; Development, ontogenetic ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunohistochemical study of the production of the intermediate filaments [vimentin, cytokeratin, and glial filament acidic protein (GFAP)] during development of the pituitary gland was made by use of fetal and adult human pituitary tissue. Among these intermediate filament proteins in the anterior and intermediate lobes of the pituitary, cytokeratin is the first to appear, followed by GFAP and vimentin. However, only cytokeratin is seen during the period of morphogenesis of the pituitary gland, with the type-II subfamily cytokeratin 8 being the earliest to appear. Among the simple-epithelial-type cytokeratins, cytokeratins 8 and 19 were observed within the pituitary primordium during morphogenesis. Cells immunoreactive for cytokeratins 8 and 19 showed a heterogeneous three-dimensional distribution pattern in Rathke's pouch. Both cytokeratins 8 and 19 tended to be strongly positive at sites in the pituitary primordium where cells had become more loosely arranged (i.e., areas far from the diencephalon) but were only weakly positive in areas in which the epithelial cells were densely packed (i.e., areas closely associated with the diencephalon). It is concluded that, during the period of morphogenesis, Rathke's pouch has the intermediate filaments characteristic of simple epithelium and shows different immunoreactivity for simple-epithelial-type cytokeratins from place to place according to the extent of cellular differentiation.
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    Morphology of the granular hemocytes of the Japanese horseshoe crabTachypleus tridentatus and immunocytochemical localization of clotting factors and antimicrobial substances (1991)
    Springer
    Cell & tissue research 266 (1991), S. 137-147 
    Details
    ISSN: 1432-0878
    Keywords: Clotting factors ; Antimicrobial substances ; Hemocytes ; Immunocytochemistry ; Tachypleus tridentatus (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The structure of hemocytes in the normal state and during blood coagulation, and the intracellular localization of three clotting factors and two antimicrobial factors were examined in the Japanese horseshoe crabTachypleus tridentatus. Two types of hemocytes were found in the circulating blood: non-granular and granular hemocytes. The latter contained numerous dense granules classed into two major types: L- and D-granules. The L-granules were larger (up to 1.5 μm in diameter) and less electron-dense than the D-granules (less than 0.6 μm in diameter). The L-granules contained three clotting factors and one antimicrobial factor, whereas the D-granules exclusively contained the other antimicrobial factor. After treatment with endotoxin, the L-granules were released more rapidly than the D-granules, although almost all granules were finally exocytosed. The granular hemocyte possessed a single Golgi complex; possible precursor granules of L-granules and D-granules contained tubular and condensed dense material, respectively. These data are discussed in relation to the self-defense mechanisms of the horseshoe crab.
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    URL: http://dx.doi.org/10.1007/BF00678720
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    Distribution of galanin-like peptide in various tissues of Necturus maculosus (1990)
    Springer
    Cell & tissue research 262 (1990), S. 461-466 
    Details
    ISSN: 1432-0878
    Keywords: Galanin ; Immunocytochemistry ; Necturus maculosus (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Galanin is a biologically active peptide which has a wide pattern of distribution in the mammalian central and peripheral nervous systems. However, the distribution of galanin-like immunoreactivity in amphibian species has not been well elucidated. In the present study, biochemical and immunohistochemical techniques were used to determine the relative concentrations, biochemical nature, and cellular localization of galanin-like immunoreactivity in the brain, heart, urinary bladder, and small intestine of Necturus maculosus (common name: mudpuppy). The results of this study indicate that each of these types of tissue contain a galanin-like peptide which is similar to porcine galanin. Brain and heart concentrations of galanin-like immuno-reactivity were particularly high, although substantial amounts were also present in the small intestine and urinary bladder. Galanin immunoreactivity was observed in ascending fiber tracts of the brainstem and in fibers in the hypothalamus. In addition, galanin immunoreactivity was observed in autonomic neurons and processes in the heart, bladder, and small intestine. The pattern of distribution of galanin-like immunoreactivity in many tissues of this amphibian species is similar to the previously described mammalian pattern; however, galanin-immunoreactive innervation of cardiac tissue has not been reported in mammals. We suggest that galanin-like immunoreactivity in the heart may be more extensive in amphibian species than in mammals.
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    URL: http://dx.doi.org/10.1007/BF00305242
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    Direct photosensitivity of chick pinealocytes as demonstrated by visinin immunoreactivity (1990)
    Springer
    Cell & tissue research 262 (1990), S. 501-505 
    Details
    ISSN: 1432-0878
    Keywords: Pinealocytes ; Visinin ; Calcium-binding protein ; Light, constant ; Photosensitization ; Immunocytochemistry ; Domestic fowl (Gallus domesticus)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Visinin, a calcium-binding protein isolated from the soluble fraction of homogenized chick retinae, has been recognized immunocytochemically in the pinealocytes of various submammals. In the chick pineal organ, continuous environmental light induced an increase in population density of visinin-immunoreactive pinealocytes. In semi-quantitative, dot-immunoblotting analysis, the amount of visinin in the pineal organs of chicks kept under continuous light for 3 days was 4–8 fold more abundant than that under continuous darkness for the same duration. Eye-enucleation and organ culture experiments clarified that this lighting effect was exerted directly on the pineal organ through the skull, and not via the neural pathway including the retinohypothalamic projection. These data suggest the existence of direct photosensitivity in the chick pinealocyte itself and the possible involvement of visinin in photoreception of the pineal organ as well as the retina of chicks.
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    URL: http://dx.doi.org/10.1007/BF00305245
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    The complex-type glycoprotein secreted by the bovine subcommissural organ: an immunological study using C1B8A8 monoclonal antibody (1991)
    Springer
    Cell & tissue research 266 (1991), S. 483-490 
    Details
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Secretory pathway ; C1B8A8 monoclonal antibody ; Immunocytochemistry ; Immunoaffinity chromatography ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The secretory pathway of the complex-type glycoprotein specific to the subcommissural organ (SCO) was examined using the monoclonal antibody (Mab) C1B8A8. Immunoreactive material was revealed in various compartments of the secretory ependymocyte, i.e., the endoplasmic reticulum, the Golgi area and the secretory vacuoles. In addition, immunoreactive material was also observed in the ventricular cavity. Evidence of a release both at the apical lining and at the basal process of the SCO ependymocytes suggests that the same protein could be secreted into the cerebrospinal fluid and the perivascular spaces. After immunoaffinity chromatography of soluble extracts of the SCO on Mab C1B8A8 immunoadsorbent columns, three glycopeptides were identified on Western blots; they were concanavalin A (Con A)-positive (88, 54 and 34 kDa) and wheat-germ agglutinin (WGA)-positive (54 and 34 kDa). The Con A-positive glycopeptide (88 kDa) is probably related to the high-mannose-type glycoprotein, the precursor form of the secreted compound, whereas the 54 kDa-glycopeptide that is both Con A- and WGA-positive could represent an intermediate form. The 34 kDa-glycopeptide that is strongly WGA-positive could be related to the monomeric form of the secreted compound. These three glycopeptides were not revealed in eluted fractions of soluble extracts of the ependyma that served as control.
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    URL: http://dx.doi.org/10.1007/BF00318589
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    Regeneration and post-metamorphic development of the central nervous system in the protochordate Ciona intestinalis: a study with monoclonal antibodies (1995)
    Springer
    Cell & tissue research 279 (1995), S. 421-432 
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    ISSN: 1432-0878
    Keywords: Key words: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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    URL: http://dx.doi.org/10.1007/s004410050299
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    Immunocytological localization of 1,25-dihydroxyvitamin D3-like molecules and their receptors in a calcium-transporting epithelium of a crustacean (1991)
    Springer
    Cell & tissue research 263 (1991), S. 345-351 
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    ISSN: 1432-0878
    Keywords: Vitamin D-1,25(OH)2D3-like immunoreactivity ; 1,25(OH)2D3 receptor-like immunoreactivity ; Immunocytochemistry ; Calcium ions ; Orchestia cavimana (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary For the first time, immunoreactivity to 1,25(OH)2D3-like molecules and their receptors has been investigated in a calcium transporting epithelium of a crustacean, Orchestia, using vertebrate antisera on ultrathin cryosections of posterior caeca previously fixed in glutaraldehyde, then postfixed in osmium tetroxide. Both immunoreactivities were located mainly in the nuclei of epithelial cells. Quantitative differences in 1,25(OH)2D3-like immunoreactivity were noticed from one stage of the molt cycle to another. These results, together with other data, contribute to evidence that immunoreactive 1,25(OH)2D3-like molecules may be involved in the regulatory processes of calcium metabolism in this terrestrial crustacean and suggest an involvement of these substances in the regulation of calcium movements in the posterior caeca.
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    URL: http://dx.doi.org/10.1007/BF00318776
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    Sulfhydryl oxidase immunoreactivity in seminiferous tubules of infertile men (1992)
    Springer
    Cell & tissue research 267 (1992), S. 209-214 
    Details
    ISSN: 1432-0878
    Keywords: Testis ; Sulfhydryl oxidase ; Hypospermatogenesis ; Sertoli cell integrity ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Sulfhydryl oxidase (SOx) immunoreactivity was investigated in the seminiferous epithelium of human biopsy material from the testes of 33 adult men with disturbed fertility. SOx immunoreactivity was expressed in normal seminiferous epithelium in type-A spermatogonia (27±4% of all spermatogonia) (n=4), in spermatocytes and round spermatids. Mature spermatozoa as well as Sertoli cells were unlabelled. within the interstitium, Leydig cells were immunopositive. In biopsies of oligozoospermic men showing hypospermatogenesis (n=24), an increase in labelled spermatogonia up to more than 90% was observed in biopsies, where seminiferous epithelia revealed only spermatogonia and Sertoli cells. Within the group of oligozoospermic patients there was a significant increase of labelled spermatogonia from 43±13% (〉20 mill/ejaculate) (n=7) to 55±16% ( 20 and 〉20 mill/ejaculate) (n=6) to 68±8% (〈5 mill/ejaculate) (n=11) and a significant (P=0.01) decrease of score count from 7.0±2.7 to 2.0±1.8. In this group the increase of labelled spermatogonia was correlated with sperm concentrations in the ajaculate (correlation coefficient: r=-0.6). In biopsies of azoospermic patients showing maturation arrest at the level of spermatocytes or spermatids (n=5) the percentage of labelled spermatogonia was within the range of 24% to 59%. Immunoreactivity in Sertoli cells was only found in single degenerating cells and in tubules showing Sertoli Cell Only Syndrome (SCO) without lumen formation. Sertoli cells within immature seminiferous cords were immunonegative, indicating that Sertoli cell SOx immunoreactivity is rather a sign of physiological alterations in degenerating cells than dependent on the stage of differentiation. Leydig cells did not show changes of immunoreactivity in any biopsy. It is concluded that SOx expression in spermatogonia may serve as a marker for spermatogenic efficiency.
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    Distribution and development of the serotonin-and RFamide-like immunoreactive neurons in the arrowworm, Paraspadella gotoi (Chaetognatha) (1992)
    Springer
    Cell & tissue research 267 (1992), S. 215-222 
    Details
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Serotonin ; RFamide ; Development ; Paraspadella gotoi (Chaetognatha)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution and development of serotonin-and RFamide-like immunoreactivities in the nervous system of Chaetognatha, Paraspadella gotoi, were examined in whole-mount preparations. In adults, a single serotonin-like immunoreactive (5HTLI) neuron and numerous RFamide-like immunoreactive (RFaLI) neurons were found in the central nervous system. Based on the structure of the fins, hooks, and eyes, seven postembryonic developmental stages were recognized. The most obvious features of the stages are: stage 1, newly hatched young; stage 2, elongation of a continuous lateral tail fin; stage 3, separation of the lateral and tail fins; stage 4, appearance of hooks; stage 5, pigmentation of eyes, stage 6, attachment by tail adhesive fins; stage 7, prey capture. Stage 1 did not show any immunoreactivity. The 5HTLI neuron first appeared at stage 4 and its axonal pathway became similar to the adult at stage 6. On the other hand, the RFaLI neurons appeared at stage 3 in the ventral ganglion. Some of their somata disappeared at stage 5 and the neuronal architecture resembled the adult at stage 7 although the RFaLI neurons in the cerebral ganglion were complete at the juvenile stage.
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    The alpha-subunit of glycoprotein hormones exists in the prolactin secretory granules of the bullforg (Rana catesbeiana) pituitary gland (1992)
    Springer
    Cell & tissue research 267 (1992), S. 223-231 
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    ISSN: 1432-0878
    Keywords: α-Subunit ; Pituitary glycoprotein hormone ; PRL cell ; Pars distalis ; Colocalization ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Our recent finding that the number of immunoreactive α-subunit cells was invariably greater than the total number of immunoreactive gonadotropin (GTH) and thyrotropin (TSH) cells in the bullfrog (Rana catesbeiana) pituitary gland raises the possibility that the α-subunit also exists in pituitary cells other than GTH and TSH cells. The present study demonstrates that there are a considerable number of immunoreactive prolactin (PRL) cells that are also stained with antibody against the α-subunit when adjacent sections are immunocytochemically examined. Neither immunoreactive growth hormone nor adrenocorticotropin cells are stained with the antibody against the α-subunit. The specificity of the antibody against the α-subunit and of that against PRL was demonstrated by preabsorption test, non-competitive binding test, and immunoblot analysis. Double-immunolabeling with gold particles of different sizes for the α-subunit and PRL revealed that most of the immunolabeled PRL-secretory granules are also labeled with the α-subunit antibody. The gold particles indicating the presence of the α-subunit were mostly found in the peripheral zone of the secretory granules.
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    Phagocytosis of lectin-opsonized fungal cells and endocytosis of the ligand by insect Spodoptera exigua granular hemocytes: an ultrastructural and immunocytochemical study (1996)
    Springer
    Cell & tissue research 285 (1996), S. 57-67 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Phagocytosis ; Insect hemocytes ; Lectins ; Fungal entomopathogens ; Ultrastructure ; Immunocytochemistry ; Cytoskeleton ; Spodoptera exigua (Insecta) ; Paecilomyces farinosus (Fungi-Deuteromycotina)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Phagocytosis of blastospores of the fungal entomopathogen Paecilomyces farinosus by granular hemocytes from larvae of Spodoptera exigua (beet armyworm) was studied. Blastospores were opsonized with a galactose-specific lectin purified from S. exigua hemolymph or with peanut agglutinin prior to incubation with hemocytes. Observations of thin sections revealed that pseudopodia extending from granulocytes attached to ligands (lectins, lectin conjugates) on the blastospores, and that the ligands became detached from the fungal surfaces and were endocytosed by granulocytes via coated pits on the plasma membrane. Coated vesicles bearing the endocytosed molecules appeared to be transported to the hemocytic granules. In other cases, ligand still coated the blastospores after phagocytosis and may have later concentrated within the phagosome along with digested fungal cell wall components. Phagocytosis of blastospores and clustering of a biotinylated lectin conjugate on or within the granulocytes were inhibited by drugs targeting cytoskeletal elements. Actin was concentrated in the pseudopodia of phagocytic granulocytes and may be directly associated with lectin receptor(s). Microtubules were abundant in the granulocytes, sometimes in specific regions.
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    Renewal of the ciliary axoneme in cone outer segments of the retina of Xenopus laevis (1996)
    Springer
    Cell & tissue research 285 (1996), S. 165-169 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Ciliary axoneme ; Cones ; Cytoskeleton ; Immunocytochemistry ; Microtubules ; Outer segments ; Photoreceptor cells ; Xenopus laevis (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The ciliary axoneme in photoreceptors from the retina of Xenopus laevis was examined by immunofluorescent staining of tubulin throughout the light/dark cycle. The immunofluorescent axoneme extended along only part of the length of rod outer segments but the entire length of cone outer segments. Both the cone axoneme and outer segment elongated during the day and shortened (presumably by shedding) during the night. Fragments of immunofluorescent axonemes were found within packets of outer segment material breaking off from cone tips. These findings show that the ciliary axoneme in the Xenopus retina is replaced during the renewal of outer segments in cones. No evidence has been found for renewal of the axoneme in rods, and thus the stability of the ciliary axoneme may differ in rod and cone photoreceptors.
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    Enkephalin-immunoreactive cells in the mesencephalic tegmentum project to the optic tectum of the teleosts Salmo gairdneri and Salmo salar (1991)
    Springer
    Cell & tissue research 264 (1991), S. 133-137 
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    ISSN: 1432-0878
    Keywords: Enkephalins ; Tegmentum mesencephali ; Tectum opticum ; Immunocytochemistry ; Retrograde labeling ; Cobalt-lysine ; Salmo gairdneri, Salmo salar (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemistry using antibodies against Met-enkephalin and Leu-enkephalin has demonstrated a group of large enkephalin-immunoreactive neurons in the nucleus of the rostral mesencephalic tegmentum (mRMT) of two teleost fish, Salmo gairdneri and Salmo salar. Injections of cobalt-lysine in the medial optic tectum retrogradely labeled the above group of tegmental neurons. Tegmental neurons were labeled only ipsilaterally to the injection site. This indicates that enkephalinergic neurons in the nRMT project to the optic tectum, and that at least some of the enkephalinergic axons observed in the optic tectum belong to a tegmento-tectal pathway. Comparable enkephalinergic pathways have been described in reptiles and birds, where pretectal-mesencephalic nuclei contribute to the enkephalin-containing fibers that project to the optic tectum.
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    Immunocytochemical colocalization of adhesive proteins with clathrin in human blood platelets: further evidence for coated vesicle-mediated transport of von Willebrand factor, fibrinogen and fibronectin (1995)
    Springer
    Cell & tissue research 279 (1995), S. 453-457 
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    ISSN: 1432-0878
    Keywords: Blood platelets ; Immunocytochemistry ; Electron microscopy ; Coated vesicles ; Clathrin ; Adhesive proteins ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Coated membranes and vesicles play an important role in receptor-mediated endocytosis and intracellular trafficking in various cell types, and are also present in blood platelets. Platelets take up certain proteins from the blood plasma, such as von Willebrand factor and fibrinogen, and these substances are transferred to storage granules. The receptors for these plasma proteins on the platelet plasma membrane have been well characterized, but morphological evidence for their transport to the storage granules is not yet available. In an attempt to clarify this aspect, we employed postembedding immunocytochemistry on platelets embedded in the acrylic resin LR White. Clathrin as the major coat component of coated vesicles was localized in the cytoplasm, on the plasmic faces of α-granules and the open canalicular system, and on the plasmic face of the plasma membrane. Colocalizations of the adhesive proteins, von Willebrand factor, fibrinogen and fibronectin, with clathrin could be observed at the same typical locations as coated vesicles were seen in Araldite-embedded material. These colocalizations have not been reported to date and furnish further evidence for a coated vesicle-mediated transport of blood plasma-derived adhesive proteins from their receptors on the outer plasma membrane to the α-granules.
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    Distribution of dopamine-like immunoreactivity suggests a role for dopamine in the courtship display behavior of the blue crab, Callinectes sapidus (1996)
    Springer
    Cell & tissue research 285 (1996), S. 321-330 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Dopamine ; Invertebrate CNS ; Immunocytochemistry ; Biogenic amine ; Callinectes sapidus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Injection of dopamine initiates a posture in the blue crab, Callinectes sapidus, identical to courtship display behavior of the male crab. The threshold for proctolin-induced rhythmic components of courtship display is lowered in preparations when dopamine is co-applied with proctolin. To elucidate the anatomical substrate of this behavior, immunocytochemistry was used to map dopamine-immunoreactive neurons. Courtship display is sex-specific, and dependent on the hormonal, developmental, and seasonal state of the animal. We compared the distribution of dopamine-like immunoreactivity between adults and juveniles of both sexes across seasons, with hormonal alteration, and with the distribution of proctolin-like immunoreactivity. Dopamine-like immunoreactivity was found throughout the nervous system in identical patterns between the sexes and hormonal states. Differences were found between juveniles and adults that are not obviously correlated with the development of behavior. Two areas of staining were of interest: neurites that longitudinally traverse and terminate in the posterior ventral nerve cord, and a neuron in the esophageal ganglion that has projections to the pericardial organ. The results do not suggest that proctolin-like and dopamine-like immunoreactivity co-localize, but in the subesophageal ganglion there was a region of close proximity.
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    GABA-immunoreactive neurons in the central and peripheral nervous system of the earthworm, Lumbricus terrestris (Oligochaeta, Annelida) (1996)
    Springer
    Cell & tissue research 285 (1996), S. 463-475 
    Details
    ISSN: 1432-0878
    Keywords: Key words: GABA (gamma-aminobutyric acid) ; Nervous system ; central ; Nervous system ; peripheral ; Immunocytochemistry ; Lumbricus terrestris (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of neurons immunoreactive for γ-aminobutyric acid was studied in the nervous system of Lumbricus terrestris (Oligochaeta). In the cerebral ganglion, the 86 cells immunoreactive for γ-aminobutyric acid represented 4.0% of the nerve cells in the brain, had a diameter of 12–50 μm, and were arranged in seven groups. Small-sized (18–30 μm) immunoreactive neurons occurred in the circumpharyngeal connectives. The axons of most immunoreactive neurons of the cerebral ganglion richly arborized in the ventral part of the neuropil and some could also be traced in the circumpharyngeal connectives. The subesophageal ganglion contained 94 immunoreactive cells (6.7% of the cells of this ganglion), also divided into seven groups, and with a diameter of 8–55 μm. The axons of the labeled neurons ran to the central neuropil giving both contra- and ipsilateral processes. Altogether 108 neurons in each ganglion (8.0% of their cells) of the ventral cord were immunopositive. Four labeled cell groups were present in the rostral and caudal part of each ganglion. Axons of these immunoreactive cells arborized in the central neuropil and projected to the segmental nerves. The stomatogastric ganglia and the enteric plexus also contained immunoreactive neurons. Many small elongated immunoreactive cells occurred in the gut epithelium. Postembedding immunogold electron microscopy revealed that immunoreactive varicosities mainly contained small pleomorphic (24 nm) agranular synaptic vesicles and some small granular (50 nm) vesicles.
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    Cellular localization of growth hormone receptors/binding proteins in immune tissues (1996)
    Springer
    Cell & tissue research 286 (1996), S. 69-80 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Growth hormone ; Growth hormone receptors ; Growth hormone-binding proteins ; Immune system ; Immunocytochemistry ; Domestic fowl (Aves ; Phasianiformes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. It is well established that the activity and proliferation of lymphoid cells and lymphoid organs are stimulated by growth hormone. These actions on lymphoid cells may be direct or mediated by actions on the epithelial and non-immune tissue cells that regulate immune function. The occurrence and cellular localization of growth hormone receptors in immune tissues has therefore been investigated to determine the target-sites of growth hormone action. Growth hormone receptor mRNA was first detected by Northern blotting in the spleen, bursa of Fabricius, and thymus of domestic fowl. In addition to the 4.4-kb transcript thought to encode the full-length growth hormone receptor, smaller transcripts of 2.8 kb and 1.0 kb, which may encode growth hormone-binding proteins, were also occasionally observed. Further analysis using the polymerase chain reaction revealed that mRNA sequences encoding the extracellular and intracellular domains of the growth hormone receptor were present in all tissues and highly homologous with hepatic transcripts. Translation of these transcripts also occurs in immune tissues, since immunoreactive growth hormone-binding proteins or growth hormone receptors of approximately 56 kDa were detected in hepatic, splenic, thymic, and bursal extracts. Immunocytochemistry of these tissues subsequently revealed that macrophages probably contain the bulk of this immunoreactivity, although some thymic medullary epithelial cells (including Hassall’s corpuscles) and splenic ellipsoids and interdigitating cells were also immunoreactive. This immunoreactivity is present in immune tissues of newly hatched and adult chickens. Importantly, B-lymphocytes were rarely, if ever, immunoreactive, and T-lymphocytes containing growth hormone receptors or binding proteins were not observed. These results suggest that a number of primary (thymus and bursa) and secondary (spleen) lymphoid tissues in the chicken contain growth hormone receptors and are thus target-sites for growth hormone action. The distribution of growth hormone receptor/growth hormone-binding protein immunoreactivity in these tissues would further suggest that growth hormone plays a major role in macrophage proliferation and/or activity and may indirectly affect lymphocyte maturation and storage via effects on thymic and splenic stromal cells.
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    GDNF triggers fiber outgrowth of fetal ventral mesencephalic grafts from nigra to striatum in 6-OHDA-lesioned rats (1996)
    Springer
    Cell & tissue research 286 (1996), S. 225-233 
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    ISSN: 1432-0878
    Keywords: Key words: Transplantation ; Fetal ventral mesencephalic grafts ; 6-Hydroxydopamine ; Glial-cell-line-derived neurotrophic factor ; Tyrosine hydroxylase ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Previous reports have indicated that grafting of fetal ventral mesencephalic tissue to the nigra region of animals unilaterally lesioned with 6-hydroxydopamine (6-OHDA), in conjunction with kainate injection between the nigra and striatum, restores nigrostriatal tyrosine hydroxylase immunoreactivity. Glial-cell-line-derived neurotrophic factor (GDNF), a potent trophic factor for dopaminergic neurons, has been found to be upregulated by kainate. We have investigated the bridging effect of GDNF injection on intra-nigral transplants. Adult Sprague-Dawley rats were anesthetized and unilaterally injected with 6-OHDA into the medial forebrain bundle. The completeness of lesions was tested by measuring methamphetamine-induced rotations. Some 1–2 months after 6-OHDA administration, fetal ventral mesencephalic tissues were grafted into the lesioned nigral area followed by injection of 100 μg GDNF, along a tract from the nigra to striatum. Animals receiving transplantation and GDNF injection showed a significant decrease in rotation 1–3 months after grafting. Immunocytochemical studies indicated that tyrosine-hydroxylase-positive neurons and fibers were present in the nigra and striatum, respectively, after grafting. No effects of similarly injected brain-derived neurotrophic factor were seen. These results indicate that fetal nigral transplantation and GDNF injection restore the nigrostriatal dopaminergic pathway in Parkinsonian animals and support the hypothesis of trophic activity of GDNF on midbrain dopaminergic neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050691
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  • 99
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    Expression and localization of GDNF in developing and adult adrenal chromaffin cells (1996)
    Springer
    Cell & tissue research 286 (1996), S. 263-268 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Chromaffin cells ; Neurotrophic factor ; RT-PCR ; Immunocytochemistry ; Western blotting ; Rat (Hannover-Wistar) ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Glial cell line-derived neurotrophic factor (GDNF) is a widely distributed member of the transforming growth factor-β superfamily and a potent neurotrophic molecule for several neuron populations in the peripheral and central nervous system. We show here that adrenal medullary chromaffin cells synthesize GDNF mRNA and contain immunoreactive GDNF protein. GDNF immunoreactivity can be found as early as embryonic day 16 in chromaffin progenitor cells of the rat adrenal gland and becomes more prominent with age. Most of the chromaffin cells within the adult rat adrenal medulla are GDNF immunoreactive, including both the noradrenergic and adrenergic subpopulations. The functions of adrenal medullary GDNF are still enigmatic but may include both auto/paracrine roles and retrograde trophic support of preganglionic neurons in the spinal cord or of sensory neurons that innervate chromaffin cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050696
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  • 100
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    Electronic Resource
    Differential expression of the EF-hand calcium-binding protein calsensin in the central nervous system of hirudinid leeches (1996)
    Springer
    Cell & tissue research 286 (1996), S. 357-364 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Calsensin ; Calcium-binding protein ; Immunocytochemistry ; Neurons ; Leeches ; Hirudo medicinalis ; Haemopis marmorata ; Macrobdella decora (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. By immunocytochemistry the distribution and developmental expression of the small EF-hand calcium-binding protein calsensin in the peripheral (PNS) and central nervous system (CNS) of the three hirudinid leech species Haemopis, Hirudo, and Macrobdella was compared. Labeling with calsensin-specific antibodies demonstrated that there was a pronounced difference in the distribution of calsensin immunoreactivity in the CNS of these leeches. In Haemopis more than 70 neurons were labeled, whereas the number in Hirudo was 51 and in Macrobdella only 8. Furthermore, the expression of calsensin in identified cells common to all three leech species also differed. Immunoblot analysis indicated that this variability was not likely to be due to multiple proteins or isoforms being recognized by the calsensin antibody. Labeling of embryos in various stages of development shows that the ontogeny of calsensin expression in the CNS is a gradual process with some neurons expressing calsensin immediately after completion of neurogenesis, about one-third of the way through embryogenesis, and others expressing calsensin only postembryonically. In contrast to the variability in the pattern and temporal expression by CNS neurons, the early embryonic calsensin expression in a small subgroup of sensillar PNS neurons was a shared feature by all three leech species. These findings suggest that calsensin may have different functional properties in CNS and PNS neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050705
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