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  • Immunocytochemistry  (352)
  • Springer  (352)
  • American Chemical Society
  • 2005-2009
  • 1985-1989  (203)
  • 1980-1984  (149)
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  • 1
    ISSN: 1432-2048
    Keywords: Cotyledons ; Endoplasmic reticulum ; Ferritin labeling ; Immunocytochemistry ; Phaseolus ; Protein (reserve) ; Reserve protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ultrastructure of the storage parenchyma cells of the cotyledons of developing bean (Phaseolus vulgaris L.) seeds was examined in ultrathin frozen sections of specimens fixed in a mixture of glutaraldehyde, formaldehyde and acrolein, infused with 1 M sucrose, and sectioned at-80° C. Ultrastructural preservation was excellent and the various subcellular organelles could readily be identified in sections which had been stained with uranyl acetate and embedded in Carbowax and methylcellulose. The cells contained large protein bodies, numerous long endoplasmic reticulum cisternae, mitochondria, dictyosomes, and electron-dense vesicles ranging in size from 0.2 to 1.0 μm. Indirect immunolabelling using rabbit immunoglobulin G against purified phaseolin (7S reserve protein), and ferritin-conjugated goat immunoglobulin G against rabbit immunoglobulin G was used to localize phaseolin. With a concentration of 0.1 mg/ml of anti-phaseolin immunoglobin G, heavy labeling with ferritin particles was observed ober the protein bodies, the cisternae of the endoplasmic reticulum, and the vesicles. The same structures were lightly labeled when the concentration of the primary antigen was 0.02 mg/ml. Ferritin particles were also found over the Golgi bodies. The absence of ferritin particles from other organelles such as mitochondria and from areas of cytoplasm devoid of organelles indicated the specificity of the staining, especially at the lower concentration of anti-phaseolin immunoglobulin G.
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  • 2
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; (PEP carboxylase) ; PEP carboxylase ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The localization of phosphoenol pyruvate carboxylase (EC 4.1.1.3.1.) in the leaf cells of Sorghum vulgare was investigated by using three techniques: the conventional aqueous and non aqueous methods gave conflicting results; the immunocytochemical techniques clearly showed that the enzyme is predominantly located in the cytoplasm of mesophyll cells.
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  • 3
    ISSN: 1432-2048
    Keywords: Cotyledon ; Golgi complex ; Immunocytochemistry ; Phaseolus (seed proteins) ; Phaseolin ; Phytohemagglutinin ; Protein (seeds)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Development of legume seeds is accompanied by the synthesis of storage proteins and lectins, and the deposition of these proteins in protein-storage vacuoles (protein bodies). We examined the subcellular distribution, in developing seeds of the common bean, Phaseolus vulgaris L., of the major storage protein (phaseolin) and the major lectin (phytohemagglutinin, PHA). The proteins were localized using an indirect immunocytochemical method in which ultrathin frozen sections were immunolabeled with rabbit antibodies specific for either PHA or phaseolin. Bound antibodies were then localized using goat-anti-rabbit immunoglobulin G adsorbed onto 4- to 5-nm colloidal gold particles. The sections were post-fixed with OsO4, dehydrated, and embedded in plastic on the grids. Both PHA and phaseolin exhibited a similar distribution in the storage-parenchyma cells, being found primarily in the developing protein bodies. Endoplasmic reticulum and Golgi complexes (cisternal stacks and associated vesicles) also were specifically labeled for both proteins, whereas the cytosol and other organelles, such as mitochondria, were not. We interpret these observations as supporting the hypothesis that the transport of storage proteins and lectins from their site of synthesis, the rough endoplasmic reticulum, to their site of deposition, the protein bodies, is mediated by the Golgi complex.
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  • 4
    Electronic Resource
    Electronic Resource
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    Planta 165 (1985), S. 522-526 
    ISSN: 1432-2048
    Keywords: Albumin (localisation) ; Cotyledon ; Pisum (albumin protein) ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The major albumin protein in storage parenchyma tissue of developing peas has been localised at an ultrastructural level by immunocytochemistry. Tissue was fixed in buffered aldehyde and embedded in LR White resin which was polymerised by addition of catalyst. Sections were labelled by the indirect method of absorption of Protein A-gold to specifically bound antibodies. This method gives high levels of specific labelling on sections which retain good ultrastructural preservation and have high contrast after conventional staining. The albumin is located throughout the cytoplasm although no labelling was found associated with the endoplasmic reticulum, Golgi apparatus, vacuoles-protein bodies or other organelles.
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  • 5
    ISSN: 1432-2048
    Keywords: Glycine (xanthine dehydrogenase) ; Immunocytochemistry ; Polyclonal antibody ; Root nodule ; Xanthine dehydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Xanthine dehydrogenase (XDH, EC 1.2.1.37) was purified from root nodules of soybean (Glycine max) and used to prepare a polyclonal rabbit antiserum. Monospecificity of this antiserum was ascertained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipate. During root nodule development of soybean, only one form of XDH was detected on an immunological basis. Titration of XDH by immunoelectrophoresis showed that a remarkable increase in the amount of XDH occurred between two and four weeks after inoculation, in parallel with the increase in enzyme activity. Localization of XDH by immunofluorescence indicated that the enzyme was present exclusively in uninfected cells where it appeared to be associated with discrete organellels
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  • 6
    Electronic Resource
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    Cellular and molecular life sciences 41 (1985), S. 1554-1557 
    ISSN: 1420-9071
    Keywords: Immunocytochemistry ; neuropeptide Y ; radioimmunoassay ; rat pancreas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Significant quantities of a newly discovered peptide, neuropeptide Y, were found in the rat pancreas, where they were localized to nerves in the exocrine parenchyma and around arterial and ductal structures. Although unaffected by surgical parasympathectomy, the periarterial and periductal nerves were abolished by chemical sympathectomy, suggesting that NPY is partially costored with sympathetic transmitters in nerve fibers.
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  • 7
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    Cellular and molecular life sciences 41 (1985), S. 1340-1342 
    ISSN: 1420-9071
    Keywords: Immunocytochemistry ; calmodulin ; secretory granules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Calmodulin is a regulator of several calcium-dependent cellular processes. It has been suggested that it plays a role in the mechanism of secretion. Employing an indirect immunoperoxidase technique at the light microscope level, this study demonstrates the presence of calmodulin in several exocytotic cells (mast cells, thyroid follicular cells, neurohypophyseal neurosecretory terminals, pancreaticβ-cells and pancreatic acinus cells) in rat and man. The positive staining reaction for calmodulin was granular and at least in the case of rat mast cells it appeared to be associated with the granule membrane.
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  • 8
    ISSN: 1432-041X
    Keywords: Vitellin ; Yolk granule ; Yolk protein ; Silkworm ; Embryogenesis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Vitellin was purified from eggs of the silkworm,Bombyx mori, by a new method in which vitellin was extracted from isolated yolk granules. The purified vitellin had a molecular weight of 540,000. An antibody against purified vitellin was prepared in rabbits. It reacted with the hemolymph vitellogenin as well as with purified vitellin, but not with other proteins in the hemolymph or in the extract from yolk granules. The anti-vitellin IgG was used to immunocytochemically locate vitellin in theBombyx non-diapause egg during early developmental stages. In the egg, just after oviposition, vitellin was located in internal yolk granules and in small yolk granules of the periplasm. During the early developmental stages studied, vitellin was not metabolized uniformly throughout the egg. The vitellin of the internal yolk granules located at the posterior-dorsal part and of the small peripheral yolk granules was utilized in 16 h and 2 days, respectively, after oviposition. A thin, very vitellin-poor layer was located between the periplasm and the vitellin-rich interior in the newly laid egg. it was always in close contact with the periphery where blastoderm and germ-band cells developed.
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  • 9
    ISSN: 1432-041X
    Keywords: Major haemolymph proteins ; Development ; Cuticle ; Immunocytochemistry ; Ceratitis capitata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The developmental profile of the major haemolymph proteins (ceratitins) inCeratitis capitata was studied. Ceratitin concentration in the haemolymph decreases dramatically during the last days of pupal life, while the amounts of ceratitins in whole organism extracts remain unchanged. By electrophoretic, immunological and immunofluorescence techniques it was revealed that ceratitins are reabsorbed by the fat body and a fraction of them is deposited in the cuticle. The possible role of ceratitins is discussed.
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  • 10
    ISSN: 1432-072X
    Keywords: Extracellular proteins ; Surface fibrils ; Algae-fungi-Chrysochromulina ; Immunocytochemistry ; Agglutination ; Fimoriae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An extensive network of extracellular fibrils was revealed by negative staining in the greenish gold algal flagellate, Chrysochromulina breviturrita. These fibrils were of uniform diameter (4–5 nm), sometimes exceeding 5 μm in length. In addition there were short, narrower fibrils (2–3 nm) on the surface of the flagella. Six protein bands were isolated from spent culture medium by SDS-PAGE and one of 80,000 Da was found to polymerize after dialysis into 4–5 nm fibrils identical to those found on the cell surface. Two other proteins of 58,000 Da and 65,000 Da also formed 4–5 nm fibrils but these were either rare or of a shorter length and different appearance. An antiserum directed against the surface 7 nm fibrils (fimbriae) of fungi agglutinated cells of C. breviturrita and some other Prymnesiophyceae and Chrysophyceae, but did not agglutinate cells of algal species in other groups. Immunofluorescence and protein A gold labelling confirmed that antigens related to fungal fimbriae were present on the surface of cells of C. breviturrita. Only the 80,000 and 58,000 Da proteins labelled heavily following protein A gold labelling. Some individual 4–5 nm fibrils labelled with gold were observed in the material prepared from the 80,000 Da band. These results therefore establish that C. breviturrita produces a surface network of fibrils that are serologically related to the fimbriae of fungi, and suggest a previously unrecognized relationship between members of the Prymnesiophyceae, Chrysophyceae and fungal groups.
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  • 11
    Electronic Resource
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    Archives of microbiology 146 (1987), S. 327-331 
    ISSN: 1432-072X
    Keywords: Actinomycetes ; Nitrogen fixation ; Symbiosis ; Immunocytochemistry ; Ultracryotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunogoldlabelling on ultrathin cryosections of Frankia sp. Cc1.17 showed specific labelling of nitrogenase in the spherical cells called vesicles. No label was found in the hyphae in any cells grown on a medium with combined nitrogen, nor in those to which no specific antiserum was added. Similar results were obtained with cultures grown under high (20%) and low (2%) oxygen tension in the gas phase.
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  • 12
    ISSN: 1432-072X
    Keywords: Hansenula polymorpha ; Peroxisomes ; Methanol ; Dihydroxyacetone synthase ; Cell fractionation ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The subcellular localization of dihydroxyacetone synthase (DHAS) in the methylotrophic yeast Hansenula polymorpha was studied by various biochemical and immunocytochemical methods. After cell fractionation involving differential and sucrose gradient centrifugation of protoplast homogenates prepared from methanol-grown cells, DHAS cosedimented with the peroxisomal enzymes alcohol oxidase and catalase. Electron microscopy of this fraction showed that it contained mainly intact peroxisomes, whereas SDS-polyacrylamide gel electrophoresis revealed two major protein bands (75 and 78 kDa) which were identified as alcohol oxidase and DHAS, respectively. The localization of DHAS in peroxisomes was further established by immunocytochemistry. After immuno-gold staining carried out on ultrathin sections of methanol-grown H. polymorpha using DHAS-specific antibodies, labelling was confined to the peroxisomal matrix.
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  • 13
    ISSN: 1432-072X
    Keywords: Methanobacterium thermoautotrophicum ; Methyl-CoM reductase ; Immunocytochemistry ; Colloidal gold ; Energy conservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Methanobacterium thermoautotrophicum were fixed with glutaraldehyde, sectioned and labeled with antibodies against the β subunit of component C (=methyl-CoM reductase) of methyl-CoM reductase system and with colloidal gold-labeled protein A. It was found that the gold particles were located predominantly in the vicinity of the cytoplasmic membrane, when the cells were grown under conditions where methyl-CoM reductase was not overproduced. This finding confirms the recent data obtained with Methanococcus voltae showing via the same immunocytochemical localization technique that in this organism methyl-CoM reductase is membrane associated.
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  • 14
    ISSN: 1432-072X
    Keywords: Acidaminococcus fermentans ; Glutamate fermentation ; Electron microscopy ; Immunocytochemistry ; Post-embedding labelling ; Antibody-gold complexes ; Protein A-gold complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the in situ location of glutaconyl-CoA decarboxylase and 2-htdroxyglutaryl-CoA dehydratase in Acidaminococcus fermentans using the antibody-gold and protein A-gold techniques carried out as a post-embedding immunoelectron microscopic procedure. Polyclonal antisera were raised in rabbits against homogeneous fractions of the enzymes. Anaerobically grown cells of A. fermentans of the late exponential growth phase were fixed with 0.2% glutaraldehyde and 0.3% formaldehyde (final concentrations) in the growth medium. Dehydration of the cells was achieved with methanol. The cells were embedded in the low temperature embedding resin Lowicryl K4M. The markers indicative for antigenic sites of the two enzymes unequivocally demonstrate that the sodium pump glutaconyl-CoA decarboxylase is located at the cell periphery being a membrane-bound enzyme as expected whereas 2-hydroxyglutaryl-CoA dehydratase is a soluble cytoplasmic enzyme.
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  • 15
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; Lectin (localization) ; Phaseolus (lectin) ; Phytohemagglutinin ; Seed (lectin)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have examined the properties and subcellular localization of phytohemagglutinin (PHA), the major lectin of the common bean (Phaseolus vulgaris.), in the axis cells of nearly mature and imbibed mature seeds. On a protein basis the axis contained about 15% as much PHA as the cotyledons. Localization of PHA was done with an indirect immunolabeling method (rabbit antibodies against PHA, followed by colloidal gold particles coated with goat antibodies against rabbit immunoglobulins) on ultra-thin cryosections which were embedded in plastic on the grids after the immunolabeling procedure. The embedding greatly improved the visualization of the subcellular structures. The small (4 nm) collodial gold particles, localized with the electron microscope, were found exclusively over small vacuoles or protein bodies in all the cell types examined (cortical parenchyma cells, vascular-bundle cells, epidermal cells). The matrix of these vacuoles-protein bodies appears considerably less dense than that of the protein bodies in the cotyledons, but the results confirm that in all parts of the embryo PHA is localized in similar structures.
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  • 16
    ISSN: 1573-5117
    Keywords: Turbellaria ; Cestoda ; Neuropeptides ; Serotonin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nervous systems of the turbellarians Microstomum lineare and Polycelis nigra and of the cestodes Diphyllobothrium dendriticum and Schistocephalus solidus were studied by means of the peroxidase-antiperoxidase (PAP) immunocytochemical method, with the use of antisera to the neuropeptides FMRF-amide, vasotocin, leu-enkephalin, met-enkephalin, neurotensin, somatostatin, and VIP, and to the bioamine serotonin. Anti-FMRF-amide positive perikarya and fibers occurred in all species, while the occurrence of the vertebrate brain-gut peptides and serotonin varied between the species. Anti-somatostatin and anti-VIP gave a negative result. Anti-FMRF-amide and anti-vasotocin positive immunoreactivity was found in the brain and gut of M. lineare, and in the CNS and the peripheral nerve net of the cestodes. We suggest that the brain-gut peptides of free-living flatworms act on the subtegumental region in the cestodes, which lack a gut but absorb their nutrients directly through the tegument.
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  • 17
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    Protoplasma 134 (1986), S. 30-42 
    ISSN: 1615-6102
    Keywords: Acetabularia ; Actin ; Cyst formation ; Immunocytochemistry ; Microtubules ; Plant cytoskeleton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cytoskeleton in the siphonous, marine green algaAcetabularia is visualized by immunocytochemistry using antibodies against plant alfa tubulin and animal smooth muscle actin. In the vegetative phase of the life cycle, when the cell grows a cylindrical stalk and until the reproductive cap is completed, actin forms continuous, parallel bundles that extend through the entire length of the stalk and cap rays respectively. Microtubules (MTs) cannot be detected until the primary nucleus, located in the rhizoid of the giant cell, divides to form thousands of secondary nuclei. MTs can then be seen radiating from each secondary nucleus that is encountered in the stalk on its migration upwards into the cap rays. They are oriented mostly parallel to the long axis of the cell. At arrival in the cap rays up to the “white spot” stage, when nuclei assume equidistant positions in the cap ray cytoplasm, a radiating system of MTs forms around each nucleus and dramatically increases until impressive radial arrays have developed. This phase coincides with a disappearance of actin bundles in the cap rays, but they are retained in the stalk cytoplasm. Shortly after that additional MTs appear around the disk like partitions of cap ray cytoplasm. Concomitantly, bundles of actin reappear colinearly with the circumferrential MTs eventually forming complete rings around each disk of cap ray cytoplasm. During this process the compartments of the future cysts are gradually bulging outwards and simultaneously the rings of actin sink inwards until domes are formed with the nuclei fixed in the top centers of the domes. At this stage the peripheral areas of the radiating MT systems around the nuclei start to break down, whereas the circumferrential MT systems remain intact. Subsequently, the rings of both actin and MTs decrease in diameter, and finally contract to a spot opposite the nucleus, while the cysts continue to develop their oval shape. After the cysts have become separated, they round up and enter several rounds of nuclear divisions. MTs form short radial arrays around each nucleus with minor changes due to a reduction of MTs during division followed by a reappearance after completion of each division. Actin is rearranged in the cysts to a cortical network of randomly oriented, short bundles, that is maintained until gamete formation sets in. These findings accentuate the involvement of Cytoskeletal elements in the key steps of morphogenesis inAcetabularia to an extent that is unknown in higher plants.
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  • 18
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Centrin ; Ca2-modulated contractile protein ; Flagellar apparatus ; Green algae ; Fibrous flagellar roots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The two main types of fibrous flagellar roots present in the flagellar apparatus of green algae (system I and system II fibers) are immunologically distinct as indicated by the localization of a Ca2+-modulated contractile protein (centrin) exclusively in one type (system II fibers) but not in the other type (system I fibers). A polyclonal antibody generated against the major protein of the striated flagellar roots (system II fibers) of the quadriflagellate green algaTetraselmis striata was used to localize centrin by immunofluorescence and pre- and postembedding immunogold electron microscopy in the flagellar apparatus ofSpermatozopsis similis, S. exsultans, Chlamydomonas reinhardtii, Dunaliella bioculata, Polytomella parva and gametes ofMonostroma grevillei andEnteromorpha sp. Whereas the antibody recognizes centrin in connecting fibers and system II fibers, no labeling occurs in system I fibers in all taxa investigated. This study presents the first evidence that system I fibers lack centrin and indicates that the two main types of fibrous flagellar roots in green algae are biochemically distinct.
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  • 19
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    Protoplasma 121 (1984), S. 25-33 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Colloidal gold ; Wheat germ agglutinin ; Lectin ; Cryosections
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chitin-binding lectin wheat germ agglutinin (WGA) is found at the periphery of wheat embryos, and a similar lectin is present at the root tips of older plants (Mishkind et al. 1982). Although a ferritin-conjugated secondary antibody is adequate for localizing WGA in embryos, native electron-opaque particles make the electron microscope identification of added label equivocal in other wheat tissues. As reported here, however, unambiguous ultrastructural localization of WGA-like lectin in adult wheat roots can be obtained with rabbit anti-WGA followed by colloidal gold-labeled goat anti-rabbit (GAR) IgG. Colloidal gold (CG) was prepared by the reduction of gold chloride with citrate, ascorbate or phosphorous. GAR IgG, prepared from serum by antigen affinity chromatograhy, was adsorbed to the gold particles to produce a stabilized suspension of GAR-CG. Localization was performed on 8–12 μM frozen sections of tissue fixed in 4% paraformaldehyde, 0.3% glutaraldehyde, and 0.75% acrolein in phosphate-buffered saline containing 1M sucrose. Localization with GAR-CG was first compared to that ascertained in embryos using other probes and was then extended to the roots of adult plants. An advantage of the GARCG method is that it permits the visualization of antigen at both the light and electron microscope levels in the same section. At the light level, the anti-WGA-GAR-CG complex appears as a red stain that is localized in specific tissues of embryos and in the caps and outer layers of adult roots. Sections in which lectin was detected at the light microscope level were embedded in plastic and sectioned for subcellular examination. Electron dense gold particles indicative of WGA are found at the periphery of protein bodies in wheat embryos and in vacuoles of the roots of adult plants. Sections incubated with control IgG lack reaction product.
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  • 20
    ISSN: 1615-6102
    Keywords: Bauhinia purpurea ; Colloidial gold ; Golgi apparatus ; Immunocytochemistry ; Lectin ; Lowicryl K4M ; Protein body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The seed lectin of the tree legume,Bauhinia purpurea alba, was localized by electron microscopic immunocytochemistry. The pattern of lectin deposition and site of intracellular localization was examined in mid- to late-maturation seeds. The seed tissue was embedded in Lowicryl K4M, the use of which with seed tissues is discussed. Immunocytochemical labeling was accomplished with colloidal gold coupled to a second antibody. The immunocytochemical reaction was specific and sensitive. Protein bodies, Golgi apparatus and Golgi secretion vesicles were densely labeled. Golgi apparatus was oriented such that Golgi secretion vesicles were in close proximity to the protein bodies. The entire Golgi apparatus was labeled with no concentration gradient across the Golgi stack. These observations indicate that the final site of lectin deposition is the protein body, and that the Golgi apparatus plays an essential role in the deposition process.
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  • 21
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    Protoplasma 144 (1988), S. 25-33 
    ISSN: 1615-6102
    Keywords: Golgi complex ; Haynaldia villosa ; Immunocytochemistry ; Prolamines ; Protein A-gold ; Protein body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Haynaldia villosa is a wild grass belonging to the tribe Triticeae, which includes important crops such as wheat, barley, and rye. The alcohol-soluble proteins ofH. villosa have extensive immunological relatedness with wheat prolamines as visualized by Western blot analysis. Amorphous protein inclusions surrounded by a limiting membrane are commonly found in the vacuoles of endosperm and subaleurone layers ofH. villosa seeds. A layer of cells just beneath the aleurone layer is rich in ER. Unlike that in other cell types, the ER in these cells is highly dilated and contains materials at its swollen distal ends. These materials are structurally similar to substances found in the protein bodies. Protein A-gold immunocytochemical localization studies employing antibodies against wheat prolamine confirmed that the inclusions found in the lumen of the ER do not contain prolamines. This observation indicates that the ER does not act as the site of prolamine accumulation inH. villosa. Protein bodies found in the vacuoles and the vesicles associated with the Golgi complexes were specifically labeled. This suggests that Golgi complexes mediate the transport of prolamines into vacuoles ofH. villosa endosperm cells, in a fashion analogous to that of other vacuolar proteins of dicotyledonous plants.
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  • 22
    ISSN: 1432-0878
    Keywords: Hypothalamo-hypophysial system ; Vasotocin neurons ; Mesotocin neurons ; Avian hypothalamus ; Immunocytochemistry ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothalamo-hypophysial system of the adult chicken has been studied with a monoclonal antibody that cross-reacts with arginine vasotocin and mesotocin. We have used this antibody on thick (100 μm) sections in conjunction with a peroxidase-conjugated rabbit antimouse antibody that permits the visualization not only of entire perikarya, but also of long portions of their axons and dendrites. Our results confirm older concepts based on classical methods, but the more sensitive immunocytochemical method reveals that the system is more extensive than previously recognized. Immunostained neurons in the chicken are widely scattered in the hypothalamus. In the rostral preoptic region, there are three immunostained neuronal cell groups: 1) a prominent closely packed group that extends along the ventromedial surface, 2) a diffusely distributed lateral group, and 3) an external group that surrounds the lateral aspect of the septomesencephalic tract. Caudally in the preoptic area and in the anterior hypothalamus, the same groups are present; but there are also conspicuous periventricular perikarya. Many of them have processes that project to the lumen of the third ventricle, as well as parallel axons that arch lateroventrally in the hypothalamus. In the midhypothalamic area, the periventricular perikarya and processes are particularly numerous at the level of the palliai commissure. The dorsal periventricular group located at the level of the dorsomedial anterior nucleus of the thalamus are the most caudal perikarya. They extend laterally in a wing-like formation. The immunostained axons from all of these perikarya form a compact hypothalamo-hypophysial tract as they run from the midhypothalamus to the median eminence and converge beneath the third ventricle. Axons branching from this tract innervate the zone externa of the anterior median eminence; another group of axons running in the fibrous layer of the zona interna proceeds to the neural lobe.
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  • 23
    ISSN: 1432-0878
    Keywords: Urophysis ; Caudal neurosecretory system ; Urotensin II ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antiserum generated against synthetic urotensin II of the goby, Gillichthys mirabilis, was used to localize urotensin II in the caudal neurosecretory system in six species of freshwater teleosts; Cyprinus carpio, Carassius auratus, Oreochromis mossambicus, Oreochromis niloticus, Salmo gairdneri and Plecoglossus altivelis, and six species of seawater teleosts: Acanthogobius flavimanus, Pagrus major, Paapristipoma trilineatum, Trachurus japonicus, Seriola dumerili and Seriola quinqueradiata. In the carp, urotensin II-immunoreactive perikarya were classified into three groups according to their size and shape. Small cells were located in the spinal cord dorsal to the urophysis, medium-sized cells immediately anterior to the urophysis, and large cells anterior to the medium-sized cells. In each group, a small number of nonreactive cells was found. Urotensin II-immunoreactive nerve fibers extended toward the urophysis and terminated around the blood vessels. Other species of teleosts showed a similar immunoreaction to that observed in the carp. The immunoreaction of the urophysis was stronger in seawater fish than freshwater fish. Urotensin II-immunoreactive elements could not be detected in the brains of the carp, goldfish and goby.
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  • 24
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Insect nervous system ; Neural development ; Calliphora erythrocephala ; Sarcophaga bullata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin-immunoreactive (5-HTi) neurons were mapped in the larval central nervous system (CNS) of the dipterous flies Calliphora erythrocephala and Sarcophaga bullata. Immunocytochemistry was performed on cryostat sections, paraffin sections, and on the entire CNS (whole mounts). The CNS of larvae displays 96–98 5-HTi cell bodies. The location of the cell bodies within the segmental cerebral and ventral ganglia is consistent among individuals. The pattern of immunoreactive fibers in tracts and within neuropil regions of the CNS was resolved in detail. Some 5-HTi neurons in the CNS possess axons that run through peripheral nerves (antenno-labro-frontal nerves). The suboesophagealand thoracico-abdominal ganglia of the adult blowflies were studied for a comparison with the larval ventral ganglia. In the thoracico-abdominal ganglia of adults the same number of 5-HTi cell bodies was found as in the larvae except in the metathoracic ganglion, which in the adult contains two cell bodies less than in the larva. The immunoreactive processes within the neuropil of the adult thoracico-abdominal ganglia form more elaborate patterns than those of the larvae, but the basic organization of major fiber tracts was similar in larval and adult ganglia. Some aspects of postembryonic development are discussed in relation to the transformation of the distribution of 5-HTi neurons and their processes into the adult pattern.
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  • 25
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Insect nervous system ; Protocerebrum ; Suboesophageal ganglion ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin-immunoreactive neurons in the median protocerebrum and suboesophageal ganglion of the sphinx moth Manduca sexta were individually reconstructed. Serotonin immunoreactivity was detected in 19–20 bilaterally symmetrical pairs of interneurons in the midbrain and 10 pairs in the suboesophageal ganglion. These neurons were also immunoreactive with antisera against DOPA decarboxylase. All major neuropil regions except the protocerebral bridge are innervated by these neurons. In addition, efferent cells are serotonin-immunoreactive in the frontal ganglion (5 neurons) and the suboesophageal ganglion (2 pairs of neurons). The latter cells probably give rise to an extensive network of immunoreactive terminals on the surface of the suboesophageal ganglion and suboesophageal nerves. Most of the serotonin-immunoreactive neurons show a gradient in the intensity of immunoreactive staining, suggesting low levels of serotonin in cell bodies and dendritic arbors and highest concentrations in axonal terminals. Serotonin-immunoreactive cells often occur in pairs with similar morphological features. With one exception, all serotonin-immunoreactive neurons have bilateral projections with at least some arborizations in identical neuropil areas in both hemispheres. The morphology of several neurons suggests that they are part of neuronal feedback circuits. The similarity in the arborization patterns of serotonin-immunoreactive neurons raises the possibility that their outgrowing neurites experienced similar forces during embryonic development. The morphological similarities further suggest that serotonin-immunoreactive interneurons in the midbrain and suboesophageal ganglion share physiological characteristics.
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  • 26
    ISSN: 1432-0878
    Keywords: Met-enkephalin-Arg6-Phe7 ; Met-enkephalin-Arg6-Gly7-Leu8 ; Immunocytochemistry ; Neuropeptides ; Co-existence of peptides ; Neurosecretory cells, insects ; Blowfly, Calliphora vomitoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal pathways immunoreactive to antisera against the extended-enkephalins, Met-enkephalin-Arg6-Phe7 (Met-7) and Met-enkephalin-Arg6-Gly7-Leu8 (Met-8), have been identified in the brain of the blowfly Calliphora vomitoria. Co-localisation with other enkephalins in certain neurons suggests that a precursor similar to preproenkephalin A exists in insects and that differential enzymatic processing occurs as in vertebrates. Co-localisations of the extended-enkephalin-like peptides with other vertebrate-type peptides, including cholecystokinin and pancreatic polypeptide, also occur. The enkephalinergic pathways are specific, comprising a few groups of highly characteristic neurons and areas of neuropil. Of special interest is the finding that parts of the antennal chemosensory and the optic lobe visual systems contain Met-8 immunoreactive neurons. Within the median neurosecretory cell groups, some of the giant neurons show immunoreactivity to Met-8 and others to both Met-8 and Met-7. Fibres from these cells project to the corpus cardiacum and also to the suboesophageal ganglion, where arborisations occur in the tritocerebral neuropil. Co-localisation studies of these cells have shown that at certain terminals, one particular type of peptide is the dominant neuroregulator, whilst at other terminals, within the same cell, a different co-synthesised peptide predominates. Several groups of lateral neurosecretory cells show clearly defined enkephalinergic pathways, most of which have connections with the central body. The complex patterns of immunoreactivity seen in terminals in the different parts of the central body, suggest an important role for the enkephalin-like peptides in the integration of multimodal sensory inputs. The physiological functions of the extended-enkephalin-like peptides in the brain of Calliphora is still unknown, but the anatomical evidence suggests they may have a role similar to that in mammals, where they are thought to control aspects of feeding behaviour.
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  • 27
    ISSN: 1432-0878
    Keywords: Gills ; Indoleamines ; Immunocytochemistry ; Autonomic innervation ; Salmo gairdneri R. ; Perca fluviatilis L. ; Micropterus dolomieui (Lacépède) ; Anguilla anguilla L. ; Ictalurus melas Rafinesque (Teleostei) ; Acipenser baeri L. (Chondrostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies against serotonin and 5-methoxytryptamine reveal indolaminergic neurons innervating the proximal part of the efferent arterial vasculature, the filament epithelia, the central venous sinus, and certain other serotonergic cells of the teleost gill filament. In the same area, acetylcholinesterase-positive and indoleaminergic neurons have already been described. We propose that these populations of neurons belong to a single neuronal type but express different agents. Our current results support this idea; in particular, they point to the presence of a single type of serotonin-containing nerve terminal, impinging on vascular smooth muscle. These results are in agreement with physiological data showing (i) the existence of non-cholinergic (atropine-resistant) vasoconstriction of the gill vasculature after nerve stimulation, and (ii) a potent vasoconstrictory action of infused serotonin. In addition, the above-mentioned serotonergic neurons have synaptic contacts with catecholaminergic nerve fibers, suggesting the existence of a modulatory relationship between the sympathetic and the cranial autonomic nerves supplying the teleost gill. Finally, these neurons show morphological relationships with a previously undescribed type of branchialserotonergic cell. The role of the parasympathetic nerve plexus of the teleost gill filament in the control of respiration and ionoregulation is discussed.
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  • 28
    ISSN: 1432-0878
    Keywords: Neurohypophysis ; Neurophysins ; Intra-axonal transport ; Immunocytochemistry ; Colchicine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Small doses (3.5 μg and 7 μg) of colchicine injected intracisternally caused an interruption of transport of secretory material from the supraoptic and paraventricular nuclei of the hypothalamus to the neural lobe of the pituitary gland. Transport was assessed by direct measurement of the incorporation of [35S] cysteine into neurophysins, by radioimmunoassay of accumulated material in discrete areas of the system and by immunocytochemistry. The larger dose (7 μg) switched off transport completely during the first 24 h but the system began to recover within three to four days. Colchicine had little, if any, effect on synthesis; comparison of the relationships of the apparent amounts of immunoreactive neurophysins and immunoreactive hormones in the arrested product led to the conclusion that processing of the hormone precursors continues within the secretory granules which accumulated in the perikarya.
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  • 29
    ISSN: 1432-0878
    Keywords: Magnocellular neurosecretory system ; Cholecystokinin ; Oxytocin-neurophysin neurons ; Immunocytochemistry ; Neuropeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The existence of cholecystokinin in the posterior hypophysis and its hypothalamic origin have been unequivocally demonstrated. Immunocytochemical evidence is presented for the coexistence of gastrin-cholecystokinin and oxytocin-neurophysin I immunoreactivities in some magnocellular neurons of the supraoptic and paraventricular nuclei both in rat and bovine hypothalamus.
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  • 30
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    Cell & tissue research 221 (1981), S. 257-267 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Human pituitary hormone antisera ; Platyfish ; Pituitary gland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this communication we describe the immunocytochemical cross-reactivity between antisera to various human pituitary hormones and specific hormone producing cell types in the pituitary gland of sexually mature male platyfish (Xiphophorus maculatus). Antisera to human pituitary hormones cross-reacted either with cells known to produce corresponding hormones (or hormone subunits) in the platyfish (e.g., ACTH, prolactin, TSH β, LH α, FSH α, TSH α) or with no pituitary cells at all (e.g., LH β, FSH β). The one exception was antiserum to human growth hormone which cross-reacted with MSH and ACTH producing cells. The platyfish pituitary is proposed as a test system for immunocytochemically screening antisera for purity and specificity in order to determine their applicability in particular studies.
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  • 31
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    Cell & tissue research 222 (1982), S. 81-100 
    ISSN: 1432-0878
    Keywords: Turtle ; Immunocytochemistry ; Pars tuberalis ; Pituitary ; Hormones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Examination of pituitaries from young and adult turtles representing four families, reveals that in addition to the abundant juxtaneural pars tuberalis (JuxPT) found in this class of reptiles, there is generally a substantial amount of pars tuberalis (PT) tissue closely associated with the pars distalis (PD). The PT forms a cortical layer especially conspicuous around the anterior tip of the PD in some species (Trionyx, Kinosternon, Sternotherus), or it forms a thick dorsal layer of tissue irregularly extending onto the sides of the PD in others (Pseudemys, Chrysemys, Lepidochelys, Chelonia). Immunocytochemical studies using unlabelled second antibody and peroxidase-antiperoxidase reveal that in turtles of all ages, the PT tissue allied with the PD (the PTinterna) is composed primarily of cells containing glycoprotein hormones (FSH, LH and TSH), especially the gonadotropins. The juxPT, however, consists mainly of secretory cells unstained by the antisera tested and includes only a small number of gonadotropes and thyrotropes. Although usually widely distributed in the testudinate adenohypophysis, the great majority of gonadotropes and thyrotropes present in the hatchling are in the PTinterna. It is probable that a concentration of these cells in the PTinterna is widespread among vertebrates. In all turtles examined, lactotropes occur principally in the anterior and ventral part of the PD proper; somatotropes are posterior and dorsal. Corticotropes are concentrated as the lactotropes in the anterior PD, but some are also scattered throughout the posterior half of the gland. Lactotropes, corticotropes, and with a few exceptions, somatotropes, do not occur in PT tissue in the turtle.
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  • 32
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    Cell & tissue research 223 (1982), S. 287-303 
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Avian brain ; Endocrine glands ; Avian embryos ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In embryos of the domestic mallard, domestic fowl, and Japanese quail vasotocin-, mesotocin-, luliberin (LHRH)-, met-enkephalin-, cortico- tropin-, and somatostatin-immunoreactive perikarya and fiber formations were visualized at different incubation stages by means of the PAP technique (Sternberger 1979). The most striking results were: (1) Vasotocin-, mesotocin-, and luliberin-immunoreactive systems display, up to the late embryonic period, morphological features most probably related to a neurohormonal function. (2) Met-enkephalin immunoreactivity appears very late during embryonic life; it is restricted to fiber networks and not found in perikarya. (3) Corticotropin immunoreactivity is observed in the tuberal region temporarily at the end of the second and the beginning of the last third of the incubation period. (4) Somatostatin-immunoreactive material is present (i) at the end of the first third of incubation, in association with the olfactory system; (ii) during the same period, adjacent to thin-layered portions of the roof of the brain; (iii) shortly thereafter, in cells of both pancreatic primordia and thyroid gland; and (iv) onward from the middle of the incubation period, in a mesencephalic cell group. The striking difference, in the early embryo, between the mature somatostatin system and the immature character of the surrounding tissues may indicate that somatostatin plays a role in the development of the brain, as well as the pancreas, and the thyroid gland.
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  • 33
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    Cell & tissue research 223 (1982), S. 695-698 
    ISSN: 1432-0878
    Keywords: Pineal gland ; Luteinizing hormone-releasing hormone (LHRH) ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactive luteinizing hormone-releasing hormone (LHRH)-like material has been demonstrated in the pineal gland of the adult rat. The objective of the present study was to examine the ontogenetic development of this LHRH-like substance in the rat pineal with the peroxidase-antiperoxidase (PAP) method of Sternberger. LHRH-like immunoreactive material was not observed in pineal glands of newborn rats. The amount of material increased progressively from the 6th–12th day of postnatal development. On day 12, the amount of LHRH-like immunoreactivity was consistent and comparable in all pineal glands of male and female animals examined.
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  • 34
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    Cell & tissue research 224 (1982), S. 25-31 
    ISSN: 1432-0878
    Keywords: Spleen ; Ig-containing cells ; Immunocytochemistry ; Germinal center ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphology and localisation of IgM- and IgG-containing cells in the spleen of rats immunized with sheep red-blood cells (SRBC) were studied by combining immunohistochemical reactions with routine histological and histochemical methods. It was shown that IgM cells occur only in the outer periarteriolar lymphocyte sheath (PALS), whereas IgG cells are present throughout the whole PALS. It has been concluded that these cell types have different routes of migration. Both IgM- and IgG-containing cells were found in the germinal center in close relation with the extracellular immune complexes. The significance of this localisation is discussed.
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  • 35
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    Cell & tissue research 224 (1982), S. 361-368 
    ISSN: 1432-0878
    Keywords: Steroids ; Pituitary ; Gonadotrops ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical reactions with anti-h(uman) TSHβ and anti-c(arp) GTHβ indicate that in juvenile rainbow trout, the dorsal basophils of the proximal pars distalis (ppd) are TSH cells, and that the ventral ones are GTH cells. Small GTH cells first appear in some fish at D(ay) 50 following fertilization (= D17 from hatching), when gonads are still undifferentiated. GTH cells increase greatly in number and size after D100. Contrary to treatment with 11β(OH)androstenedione, addition of methyl testosterone, progesterone and 17α(OH)progesterone stimulate the development of granular GTH cells in rainbow trout, shortly after hatching. It is suggested that steroid receptors in the brain or pituitary discriminate between 11-steroids and other steroids rather than between aromatizable androgens and other steroids.
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  • 36
    ISSN: 1432-0878
    Keywords: Vertebrate-type neuropeptides ; Neuroendocrine system of insects ; Neurosecretion ; Peptidergic neurons ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and differential distribution of substances antigenically related to known vertebrate neuropeptides demonstrated within the corpus cardiacum of the insect Leucophaea are as follows: Of ten mammalian antisera tested, six yielded substantial immunoreactive deposits resembling oxytocin, somatostatin, Substance P, met-enkephalin, bombesin, and neurotensin, respectively. In the remaining four, the reaction was moderate (vasopressin, β-endorphin) or marginal (LH-RF, calcitonin). With regard to their regional distribution, these biochemically distinct reaction products seem to fall into two groups: (1) Materials resembling oxytocin, vasopressin, met-enkephalin, β-endorphin (and presumably also neurotensin and LH-RF) predominate in the central release area of the organ and are considered to be of extrinsic (cerebral) origin. (2) Substances localized primarily in areas rich in intrinsic glandular cells of the corpus cardiacum, and revealed by antisera raised against somatostatin, Substance P, and bombesin, are judged to be synthesized and stored within this organ. In peptidergic fibers entering the adjacent corpora allata, thus far Substance P-, β-endorphin-, and LH-RF-like immunoreactivities have been demonstrated. Some of these “new” neuropeptides may be contained in classical neurosecretory neurons, formerly identified by less specific methods, others must be assigned to additional peptidergic neurons heretofore unknown.
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  • 37
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Cardioactive peptide ; Lymnaea stagnalis ; Neurohormone ; Neurotransmitter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With an antiserum to the molluscan cardioactive tetrapeptide FMRF-amide immunoreactive perikarya and nerve fibers were identified in the central and peripheral nervous system of the pond snail Lymnaea stagnalis. Their localization is described. The same antiserum yielded reactive product in particular cells of the epithelium of the alimentary tract. The use of two different fixatives, glutaraldehyde, and a mixture of glutaraldehyde, picric acid, and acetic acid (GPA) showed that certain nerve cells can be identified only in material fixed with either the one or the other of these two fixatives, a result which indicates that in Lymnaea more than one FMRF-amide-like substance may occur. “Positive” axon endings were found in the periphery of various nerves, i.e., in places where neurohormones are released into the blood. Other fibers were found to end, probably synaptically, on other neurons, on epithelial cells in the stomach, and between muscle cells in various parts of the body, e.g., in the heart. In these cases the FMRF-amide-like substance may function as a neurotransmitter or a neuromodulator.
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  • 38
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide (PP)-like material ; Immunocytochemistry ; Nervous system ; Periplaneta americana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The central and visceral nervous systems of the cockroach Periplaneta americana were studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibody to bovine pancreatic polypeptide (PP). PP-like immunoreactive neuron somata are most numerous in the brain; at least 6 pairs of cell groups occur in clearly defined regions. Three pairs of cells each are also present in the suboesophageal ganglion and the thoracic ganglia, one pair of a single cell each in the first abdominal and the frontal ganglia, and 4 to 6 pairs of single cells in the terminal ganglion. No reactive cells were found in the retrocerebral complex and the second to the fifth abdominal ganglia. The axons containing PP-like immunoreactivity issue many branches that are distributed in the entire brain-retrocerebral complex, ventral cord, and visceral nervous system. PP-like immunoreactive material produced in the brain seems to be transported by three routes: protocerebrum to corpora cardiaca (-allata) through the nervi corporis cardiaci, tritocerebrum to visceral nervous system through frontal commissures, and to ventral cord through circumoesophageal connectives. A possible homology between the mammalian brain-GEP (gastro-enteropancreatic) system and the brain-midgut system of this insect is discussed.
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  • 39
    ISSN: 1432-0878
    Keywords: Pituitary ; Adenohypophysis ; Luteinizing hormone (LH) ; Immunocytochemistry ; Lamprey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A mammalian-like immunoreactive luteinizing hormone (LH) has been localized in the cytoplasm of cells in the meso-adenohypophysis of the sea lamprey, Petromyzon marinus L. Specific staining for LH was absent in other regions of the lamprey pituitary.
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  • 40
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    Cell & tissue research 229 (1983), S. 541-550 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Hagfish ; Somatostatin neurons ; Vasotocin neurons ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The brain of the Pacific hagfish, Eptatretus stouti, was studied immunocytochemically using antisera against somatostatin (SRIH), arginine vasopressin (AVP), and adrenocorticotropic hormone (ACTH). SRIH-immunoreactive perikarya were distributed bilaterally in the postoptic nucleus and in the hypothalamic nucleus. Although several short, stained fibers were observed in the vicinity of the perikarya, SRIH-immunoreactivity was not found in the neurohypophysis, nor in other parts of the brain. On the other hand, presumed arginine vasotocin (AVT) perikarya were distributed in an arc-shaped region extending from the posterior part of the preoptic nucleus to the anterior-most end of the hypothalamic nucleus and projected their fibers to the neurohypophysis. Most presumptive AVT perikarya were located close to the paired prehypophysial arteries near the anterior end of the postoptic nucleus. In the neurohypophysis, abundant presumptive AVT-fibers terminated in the posterior dorsal wall, although some fibers terminated in the anterior dorsal wall and only a few fiber endings were found in the ventral wall. No ACTH-positive cells were detected in the hagfish brain or in the pituitary gland.
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  • 41
    ISSN: 1432-0878
    Keywords: Somatotrops ; Oncorhynchus ; Immunocytochemistry ; Immunodiffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary somatotrops were localized in four species from the genus Oncorhynchus using an antiserum to chum salmon (Oncorhynchus keta) growth hormone. The antiserum cross-reacted specifically with the acidophils in the caudal pars distalis of all species tested. Corroborative immunodiffusion studies were also conducted with pituitary homogenates from the same species. Immunological identity of the GH molecule was observed for all species tested.
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  • 42
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    Cell & tissue research 233 (1983), S. 99-111 
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Vasopression ; Osmotic stress ; RIA ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A correlative radioimmunoassay (RIA) and immunocytochemical (ICC) study was carried out on vasopressin (VP) distribution and content in brains of normal and 3-day water-deprived rats. By RIA there were statistically significant differences in brain VP per pg/mg between normal and osmotically stressed specimens in hypothalamus (338.4 versus 134.4), thalamus (4.8 versus 0.9), septum (18.0 versus 3.4), striatum (1.6 versus 0.7) and amygdala (17.3 versus 1.3), but not in other brain regions measured. Pituitary VP decreased from 71.1 to 8.7 ng/mg, and plasma VP rose from 3.6 to 19.3 pg/ml during water deprivation. Application of the peroxidase-anti-peroxidase ICC method of Sternberger to vibratome sections showed that VP-immunoreactivity in dehydrated specimens decreased in perikarya of paraventricular nucleus and suprachiasmatic nucleus, while intrahypothalamic immunoreactive magnocellular fibers appeared more conspicuous due to proliferation of large Herring bodies. In extrahypothalamic sites VP-immunoreactivity in water-deprived rats was visibly reduced in periventricular thalamus and septum. Thus it is apparent that both intra- and extrahypothalamic VP are affected by osmotic stress, and these results are discussed within the context of current ideas relating to co-activation of neurosecretory cells that project to different sites.
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  • 43
    ISSN: 1432-0878
    Keywords: Lung development ; Fibronectin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of the rat lung is a process of continuing morphological change. Indications from work in other mammalian systems suggest that fibronectin may be important in the control of this process. The present study has examined embryonic, neonatal, and adult lung tissue of the rat by means of the peroxidase-antiperoxidase (PAP) technique to demonstrate fibronectin at the light-microscopic level. Positive reaction was observed with anti-fibronectin serum in all stages examined. Control sections treated with pre-immune serum or no primary serum gave negative results in each case. Fibronectin in adult tissue was localized to the alveolar surface and alveolar basal lamina. Neonatal tissue showed fibronectin on pulmonary tubule walls and in basal lamina while embryonic tissue revealed localization of the protein in the basal lamina and in association with small groups of cells at the base of septal buds. These findings suggest a role for fibronectin in the control of rat lung development. The results are discussed in terms of the known functions of fibronectin as a preliminary matrix for the subsequent deposition of collagenous connective tissue, as a cellular adhesion protein, and as surface-bound material for cellular migration.
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  • 44
    ISSN: 1432-0878
    Keywords: Mesotocin neurons ; Vasotocin neurons ; Avian hypothalamus ; Preoptic area ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The neurosecretory systems producing mesotocin (MT) and vasotocin (VT) (the avian homologues of oxytocin and vasopressin, respectively) were characterized in the brains of the domestic mallard and Japanese quail by means of indirect immunofluorescence techniques using specific antisera. In the anterior preoptic region, including the organum vasculosum of the lamina terminalis, and at different levels of the supraoptic and paraventricular nuclei, separate mesotocin- and vasotocin-producing neurons were identified. Mesotocinergic and vasotocinergic neurons were also located in the tuberomammillary area, among the ectomammillary tract fibers. The supraoptico-neurohypophysial tract, formed by vasotocin- and mesotocincontaining axons, enters the internal zone of the median eminence and ends in the posterior lobe of the pituitary. The external zone of the rostral median eminence appears to contain vasotocin and mesotocin fibers, which terminate in close contact with the capillaries of the hypophysial portal system.
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  • 45
    ISSN: 1432-0878
    Keywords: Teleost ; Pars intermedia ; Immunocytochemistry ; PAP-technique ; MSH ; ACTH ; Endorphin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pars intermedia of S. mossambicus contains two different endocrine-cell types. The predominant cell type is lead-haematoxyline-positive and assumed to synthesize MSH and related peptides. The second cell type is PAS positive and its function and product(s) are unknown. Staining of light-microscopic and ultrathin sections with antisera against α-MSH, ACTH 1–24 and human β-endorphin revealed that only the lead-haematoxyline-positive cells of the pars intermedia react with these antisera, and that the secretory granules of these cells contain compounds that were immunoreactive to all three antisera. These findings are in line with the hypothesis that α-MSH, ACTH and endorphins are derived from the same precursor molecule. No specific reaction with one of the antisera could be detected in the PAS positive cells.
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  • 46
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    Cell & tissue research 233 (1983), S. 389-402 
    ISSN: 1432-0878
    Keywords: Calcium-binding protein (CaBP) ; Immunocytochemistry ; Rat cerebellum ; Development ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific antibodies raised against a human 28 000 dalton cerebellar calcium-binding protein (CaBP) were used in an immunocytochemical study during development of the rat cerebellum. Both light and electron microscopy showed (1) that labelling was entirely restricted to the Purkinje cells, (2) that it appeared very early in Purkinje cell development, (3) that the entire cell was labelled from the tip of the smallest dendrites to the axonal terminals, and (4) that with increasing age, the immunoreaction appeared to be progressively restricted to the cell and organelle membranes.
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  • 47
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biologically active peptides ; Lymnaea stagnalis ; Neurohormone ; Neurotransmitter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Perikarya and nerve fibers were immunocytochemically identified in the central nervous system of the pond snail Lymnaea stagnalis by means of the unlabelled antibody enzyme method with antisera to 15 biologically active peptides of vertebrates: vasopressin, vasotocin, oxytocin, α-melanocyte stimulating hormone (α-MSH), met-enkephalin, somatostatin, glucagon, insulin, glucose-dependent insulinotropic peptide (GIP), vaso-active intestinal polypeptide (VIP), gastrin, secretin, pancreatic polypeptide (PP), Substance P, calcitonin. No immunostaining was obtained with antisera to β-endorphin, cholecystokinin (CCK), neurophysin I and II. Particular neurons could be identified with two antisera (anti-vasopressin/vasotocin, anti-α-MSH/metenkephalin, anti-substance P/PP, anti-PP/gastrin). Apparently this indicates that populations of cells identified with a given antiserum may consist of more than one cell type. Only a few of the new peptidergic cells appeared to be identical with classical neurosecretory cells. Thus the growth hormone producing Light Green Cells stained with anti-somatostatin and the axon terminals of the ovulation hormone producing Caudo-Dorsal Cells with anti-met-enkephalin. Whether this indicates structural identity of the growth hormone with somatostatin and of the ovulation hormone with met-enkephalin remains to be investigated. Just like the classical neurosecretory cells a number of the new peptidergic cells (anti-glucagon, -insulin, -met-enkephalin, -somatostatin, and -PP positive cells) send their axons to the peripheries of commissures, connectives or nerves. Thus these cells can be considered as probably neuroendocrine. The classical neurosecretory cells release their products into the haemolymph from these sites. Other new peptidergic cells (e.g., anti-vasopressin, -vasotocin, -oxytocin and -GIP positive cells) have axons that terminate, probably synaptically, on other neurons, indicating that they are “more conventional” neurons, their products being neurotransmitters/neuromodulators. It can also not be excluded that some cells of a population containing a given peptide are neuroendocrine and others make contact with other neurons.
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  • 48
    ISSN: 1432-0878
    Keywords: Digestive tract ; Fish ; Gastrin-immunoreactive cells ; Pancreatic polypeptide-immunoreactive cells ; Somatostatin-immunoreactive cells ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Gastrin, pancreatic polypeptide and somatostatin immunoreactive cells in the gut of two fish with stomachs (perch and catfish) and a stomachless fish (carp) were studied by immunocytochemistry. In the gastric mucosa of perch and catfish, cells showing gastrin and somatostatin-like immunoreactivity are found, scattered among the surface mucous cells and mucous neck cells. No pancreatic polypeptide (P.P.) immunoreactive cells are detected in the gastric mucosa. Cells showing gastrin and P.P.-like immunoreactivity are observed in the intestinal mucosa of perch, catfish and carp. In this location no somatostatin immunoreactive cells are found.
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  • 49
    ISSN: 1432-0878
    Keywords: Urotensin ; Caudal neurosecretory system ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of urotensin I has been investigated in the caudal neurosecretory system of the white sucker (Catostomus commersoni). The peptide is present in all the cells of the system both large and small, in the large axons passing to the urophysis, and in fine beaded fibres not only within the urophysis but also in a fine plexus lateral to the large cells in the spinal cord proper. The possibility that the caudal neurosecretory system is not a functionally uniform system but rather a collection of dissimilar cells of different synaptic inputs with a common entity, urotensin I, is discussed. Moreover, the feasibility of a urotensin I feedback loop is described.
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  • 50
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    Cell & tissue research 235 (1984), S. 211-214 
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Pars distalis ; Immunocytochemistry ; Amphibia ; Gymnophiona
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunofluorescence method was used to identify and locate LTH-, STH-, LH-, TSH-, ACTH- and MSH-immunoreactive cells in the pituitary of Typhlonectes compressicaudus (Gymnophiona). The present study defines the histological and histochemical staining properties of each cell type identified.
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  • 51
    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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  • 52
    ISSN: 1432-0878
    Keywords: FMRFamide ; Bovine pancreatic polypeptide ; Immunocytochemistry ; Peptidergic neurons ; Leptinotarsa decemlineata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Particular neurons in the nervous system of the Colorado potato beetle, Leptinotarsa decemlineata, are recognized by antisera against bovine pancreatic polypeptide and FMRFamide. Both antisera react with the same neurons. Solid phase absorptions showed that antiserum against bovine pancreatic polypeptide cross-reacts with FMRFamide, whereas antiserum against FMRFamide cross-reacts with bovine pancreatic polypeptide. Some of the immunoreactive neurons have axons branching extensively within the neuropile, which suggests that the peptide is used as transmitter. In the corpus cardiacum, a neurohaemal organ in insects, numerous immunoreactive axon terminals are present. Here, the peptide material is presumably released as a hormone.
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  • 53
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    Cell & tissue research 235 (1984), S. 427-432 
    ISSN: 1432-0878
    Keywords: DPP IV ; Salivary glands ; Ultrastructural localization ; Immunocytochemistry ; PAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of dipeptidyl peptidase IV (DPP IV) (EC 3.4.14.5) in rat submandibular and parotid glands was studied immunocytochemically by the peroxidase-antiperoxidase (PAP) method, using a monospecific antiserum against rat kidney DPP IV. There were no differences in the immunocytochemical localization of DPP IV between submandibular and parotid glands. In these glands, DPP IV was primarily found to be associated with the luminal and intercellular canalicular plasma membranes of acinar cells and with the luminal plasma membranes of intercalated and striated duct cells. Occasionally, immunoreaction of DPP IV was detected in cytoplasmic vesicles (vacuoles), lysosomes, and multivesicular bodies in some acinar cells as well as in ductal epithelial cells. Furthermore, the reaction product was also found within the lumina of peri-acinar and peri-ductal capillaries and in the cytoplasm of some fibroblasts in the interstitial connective tissue. These data suggest that DPP IV in the submandibular and parotid glands may play some role in the secretion or reabsorption processes of secretory proteins and peptides in these glands.
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  • 54
    ISSN: 1432-0878
    Keywords: S-100 protein ; Thymus ; Interdigitating cells ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study deals with the localization and ultrastructure of S-100-immunoreactive cells in the human thymus. These immunoreactive cells are distributed mainly in the medulla with some scattered elements in the cortex. Electron-microscopic observation revealed that the cells are characterized by an irregularly shaped nucleus, tubulovesicular structures in the cytoplasm and characteristic interdigitations of the plasma membrane. The cells often embrace lymphocytes with their branched processes. On the basis of these morphological features, the immunostained elements were identified as interdigitating cells (IDCs). The immunocytochemistry for S-100 visualizes the precise distribution and extension of the IDCs under the light microscope and indicates that the IDCs form no structural networks such as those established by the thymic epithelial cells. Since the IDCs in human lymph nodes have also been reported to contain S-100-like immunoreactivity, S-100 protein can be regarded as a useful marker for identifying the IDCs in the human thymus and other lymphoid organs.
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  • 55
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    Cell & tissue research 246 (1986), S. 205-210 
    ISSN: 1432-0878
    Keywords: 1,2-Dimethylhydrazine ; Peptide YY ; Glucagon ; 5-Hydroxytryptamine ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Colorectal adenocarcinomas were induced in male Wistar rats, by weekly subcutaneous administration of 1,2-dimethylhydrazine, classified according to the degree of differentiation and submitted to immunocytochemistry for the peptides cholecystokinin (CCK), gastrin, gastric inhibitory polypeptide (GIP), glucagon, neurotensin, pancreatic polypeptide (PP), peptide YY (PYY), somatostatin and vasoactive intestinal polypeptide (VIP) and the biogenic monoamine 5-hydroxytryptamine. Well- or moderately well-differentiated adenocarcinomas comprised 46% of the tumour population, only 4% were poorly-differentiated adenocarcinomas, and the remaining 50% possessed a mixture of these two morphologies. Glucagon, PYY and 5-hydroxytryptamine immunoreactive cells were frequently observed within well- or moderately well-differentiated tumours and within such regions of tumours possessing a mixed morphological pattern. The tumours contained no cells immunoreactive for any of the peptides not normally located within the colorectum, nor did they contain cells immunoreactive for somatostatin and VIP, although known positive controls did stain. Poorly-differentiated tumours and portions of tumours of mixed type, were consistently negative. 5-hydroxytryptamine was the most frequently located of the three antigens, being detected in 87% of the moderately well-differentiated tumours and 32% of the tumours with mixed morphologies. 11% of moderately well-differentiated tumours possessed 5-hydroxytryptamine positive cells in such profusion that they contributed significantly to the tumour mass. The distribution of glucagon-and PYY-immunoreactive cells was similar, although they occurred with a lower frequency, presumably corresponding to their lower numbers within the normal colorectal mucosa. Additionally, these two peptide immunoreactivities were colocalized in the majority of cells, although some cells contained only one antigen. The immense numbers of cells immunoreactive for peptides and monoamine in a significant proportion of colorectal adenocarcinomas suggests that they have arisen from multipotential endodermal stem cells within the tumours and are not part of the normal epithelial population being engulfed as the tumour grows.
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  • 56
    ISSN: 1432-0878
    Keywords: Parathyroid ; Storage granule ; Hypercalcemia ; Immunocytochemistry ; Acid phosphatase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Short-term effects of CaCl2-treatment on parathyroid cells of the rat, especially on their storage granules, were studied at the ultrastructural level. After an injection of 4% CaCl2, serum calcium levels (SCL) rapidly increased from 9.1 mg/dl (controls) to a maximum of 14.9 mg/dl at 20 min. At 5 min after the injection, the number of type-I storage granules (large core) [NSG-I] and that of type-II storage granules (small core) [NSG-II] remained unchanged, in spite of elevated SCL (12.4 mg/dl). As soon as SCL rose to 13.2 mg/dl at 7.5 min, NSG-I gradually decreased to a minimum at 30 min; in contrast, NSG-II gradually increased to a maximum at 30 min. Vacuolar bodies also increased together with the augmentation of type-II storage granules. The average diameter of the core of the storage granules decreased significantly after the injection. Protein A-gold method for immunocytochemistry showed that the cores of these granules contain parathormone. Acid-phosphatase activity was occasionally found in storage granules of both types, especially in those of type II. It is concluded (i) that type-I storage granules may be transformed into vacuolar bodies via type-II granules as a result of hydrolysis, and (ii) that these processes may be accelerated during hypercalcemia.
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  • 57
    ISSN: 1432-0878
    Keywords: Denervation ; Cholinergic synaptic vesicles ; Proteoglycan ; Immunocytochemistry ; Immuno-electron microscopy ; Torpedo marmorata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A proteoglycan-specific antiserum has been used to monitor the effects of denervation in the electric organ of Torpedo marmorata. The antiserum was produced by injecting a highly purified synaptic vesicle fraction prepared from the electric organs of Torpedo marmorata. Following absorption the serum appears to be specific towards synaptic vesicles. The ultrastructural localization of the antigen determined by immuno-electron microscopy confirmed the specificity of the antiserum and showed that it did not crossreact with the proteoglycans of the basal lamina. The rate of disappearance of the vesicle proteoglycans following denervation was evaluated by means of the antiserum and was compared to the rate of disappearance of other vesicular and nerve terminal-associated markers. The results suggest that degeneration affects the vesicular constituents at varying rates resulting in a progressive disappearance of the entire functional capacity of the synaptic vesicles.
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  • 58
    ISSN: 1432-0878
    Keywords: Fish skin ; Sacciform glandular cells ; Immunocytochemistry ; 5-Hydroxytryptamine ; Lepadogaster candollei
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin has been demonstrated in the epidermal sacciform glandular cells of the clingfish Lepadogaster candollei by use of immunocytochemistry. Serotonin immunoreactivity is found both in the peripheral cytoplasm of the glandular cells and their luminal secretion. The presence of serotonin in the sacciform glandular cells parallels that located by both biochemical and immunocytochemical procedures in the cutaneous glands of many amphibian species.
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  • 59
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biologically active peptides ; Peptidergic neurons ; Crayfish ; Astacus leptodactylus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The peptidergic system in the optic ganglia of Astacus leptodactylus is characterized by the immunocytochemical application of 15 antisera raised against biologically active peptides of vertebrates and invertebrates. Positive reactions were found with anti-FMRFamide, antiαMSH, anti-vasotocin, anti-gastrin, anti-CCK, anti-oxytocin, anti-secretin, anti-glucagon and anti-GIP. Based on immunochemical reaction and localization it is possible to distinguish 30 cell groups. Only part of these cell groups is found in the known classical neurosecretory cell regions. This observation demonstrates a more extensive peptidergic system than formerly recognized. The morphology of this peptidergic system suggests that one part is neurohormonal and the other part neurotransmitter-like or neuromodulatory.
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  • 60
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone (MCH) ; Nucleus lateralis tuberis ; Hypothalamo-hypophysial relationship ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Melanin-concentrating hormone (MCH) has been purified from the chum salmon pituitary. Its complete amino acid sequence has recently been established. To identify the precise site of origin of MCH, immunostaining was performed in the brain and pituitary gland of the chum salmon and the rainbow trout using a highly sensitive and specific antiserum raised against synthetic MCH. In these two salmonid species immunoreactivity for MCH was detected in neurons and neuronal processes in the pars lateralis of the nucleus lateralis tuberis (NLT) in the basal hypothalamus. Numerous positive-staining processes of these MCH-neurons project to the pituitary gland, extending into neurohypophysial tissues within the pars intermedia and, to a lesser extent, into the pars distalis. No pituitary cells showed cross-reactivity. These results suggest that MCH is biosynthesized in the neurons of the NLT/pars lateralis and released in the neurohypophysis. On the other hand, prominent but less numerous MCH-positive processes could be traced to the pretectal area in which projection of both optic and pineal fibers has been detected using tracers. This observation suggests that the synthesis and/or release of MCH might be under the influence of either of these photosensory neurons. Moreover, the existence of an extrahypothalamic projection from MCH-positive neurons suggests that, in addition to melanin-concentration, MCH might be involved in other neuronal functions, perhaps serving as neuromodulator in the brain.
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  • 61
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    Cell & tissue research 240 (1985), S. 497-500 
    ISSN: 1432-0878
    Keywords: α-MSH-like peptides ; Immunocytochemistry ; Neural gland ; Ascidian (Styela plicata) ; Pituitary-like role
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An α-MSH-like immunoreactivity has been localized in the neural gland of the ascidian Styela plicata. In particular, immunoreactive cells occur in some lobules and are weakly lead-haematoxylin positive. On the basis of the results, the homology of the ascidian neural gland with the vertebrate adenohyphysis is suggested and discussed. Furthermore, some hypotheses are presented about the possible functions of α-MSH-like material in ascidians.
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  • 62
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    Cell & tissue research 247 (1987), S. 249-258 
    ISSN: 1432-0878
    Keywords: Lactoferrin ; Electron microscope ; Immunocytochemistry ; Neutrophils ; Morphometry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcellular localization of lactoferrin in human neutrophils was studied by an electron-microscopic immunoperoxidase method. This molecule was detected in small granules of blood polymorphonuclear leukocytes. A morphometrical analysis showed that there was no significant difference in the mean size between lactoferrin-positive and myeloperoxidase-negative granules. In contrast, the mean size of myeloperoxidase-positive granules was significantly larger than that of lactoferrin-positive granules. This indicates that lactoferrin is contained in the myeloperoxidase-negative, secondary, granules of human neutrophils. In immature bone marrow mononuclear neutrophils, lactoferrin was present in cytoplasmic granules of somewhat larger size than lactoferrin-positive granules of polymorphonuclear leucocytes. A morphometrical study showed that the mean size of lactoferrin-positive granules was significantly greater in immature bone marrow cells than in polymorphonuclear leucocytes. This indicates that lactoferrin-positive granules decrease in size as the cells mature. Besides cytoplasmic granules, lactoferrin was demonstrated in the Golgi complex and a part of the rough endoplasmic reticulum of immature bone marrow neutrophils, probably myelocytes and early metamyelocytes. These results show that lactoferrin is synthesized and packed into secondary granules in immature bone marrow neutrophils and therefore that the secondary granules are a type of secretory granule.
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  • 63
    ISSN: 1432-0878
    Keywords: Nucleus preopticus ; Nucleus lateralis tuberis ; Corticotropin-releasing factor (CRF) ; Arginine vasotocin (AVT) ; Immunocytochemistry ; Catostomus commersoni (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The peroxidase-antiperoxidase immunocytochemical procedure was used to study the distribution of ovine corticotropin-releasing factor (CRF) and arginine vasotocin (AVT) immunoreactivities sequentially in the same sections or in adjacent sections of the brain and pituitary of Catostomus commersoni. It was found that all CRF-immunoreactive (IR) neurons in the nucleus preopticus (NPO) also contained AVT immunoreactivity. Co-localization of both immunoreactivities was also observed in fibres forming the preoptic-pituitary tract and in the neurohypophyseal digitations, the IR-CRF and IR-AVT fibres projecting mainly to the neurointermediate lobe (NIL) of the pituitary. An additional population of exclusively IR-AVT neurons and fibres in the NPO, preoptic-pituitary tract and NIL was also observed. Exclusive CRF-immunostaining was found in neurons of the nucleus lateralis tuberis (NLT), in fibres distributed in some diencephalic nuclei and in the neurohypophyseal digitations in the region of the rostral pars distalis (RPD). These results suggest (i) that CRF- and AVT-like substances, present in NIL fibres (probably originating in the NPO), may have an integrated role in the release of the cell products from the pars intermedia, and (ii) that the control of corticotrops in the rostral pars distalis, innervated exclusively by IR-CRF fibres (probably originating in the NLT), does not require a simultaneous presence of CRF- and AVT-like substances.
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  • 64
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide ; FMRFamide ; Retina ; Immunocytochemistry ; Carassius auratus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical studies were conducted on goldfish to determine whether a retinal efferent fiber system, immunoreactive to the tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRFamide), might contain instead a substance similar to one of the 36-amino acid pancreatic polypeptides, the C-terminus of which is similar to FMRFamide. Our results demonstrate the presence of two separate peptidergic systems, one containing FMRFamide-like, and the other pancreatic polypeptide-like peptides. Antisera to FMRFamide reveal the efferent fibers, whose axons exit the optic nerve and terminate in layer 1 of the inner plexiform layer, as previously described. Antisera to porcine neuropeptide Y, and to avian and bovine pancreatic polypeptides label a sparse population of putative amacrine cell bodies and a dense fiber plexus in layers 1, 3, and 5 of the inner plexiform layer. Based on intensity of staining, this amacrine cell peptide appears to be most similar to neuropeptide-Y. Radioimmunoassay and immunocytochemical staining of retinas in which the efferent fiber peptide was depleted by optic nerve crush confirm in large part the observation that the two peptide systems are distinct. However, there is some cross-recognition of the FMRFamide-like tissue antigen by pancreatic polypeptide antibodies. Double-label studies with antisera to tyrosine hydroxylase and neuropeptide-Y indicate that the pancreatic polypeptide antigen is not co-localized with catecholamines.
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  • 65
    ISSN: 1432-0878
    Keywords: Reissner's fiber ; Massa caudalis ; Subcommissural organ ; Spinal cord ; Central canal ; Immunocytochemistry ; Lectin histochemistry ; Lamprey ; Geotria australis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO), Reissner's fiber (RF) and its massa caudalis of lamprey larvae (Geotria australis) were investigated immunocytochemically by use of an antiserum raised against bovine RF as primary antibody. The affinities of RF and massa caudalis for Ricinus communis agglutinin I (RCA) with and without previous acid hydrolysis, concanavalin A (Con A), wheat-germ agglutinin (WGA), aldehyde fuchsin, and PAS reaction were also studied. SCO and massa caudalis were strongly immunoreactive, whereas RF proper was distinctly negative. RF did not react with Con A and RCA. Only the periphery of RF was WGA-positive. RCA showed affinity for RF only after acid hydrolysis. RF was homogeneously stained by the aldehyde-fuchsin and PAS-methods. At variance with RF proper, the periphery of the massa caudalis reacted with RCA without previous acid hydrolysis, but its core was WGA-positive and reacted with RCA only after hydrolysis. It is suggested that (i) RF has a coat of glycoproteins containing sialic acid as terminal residue, whereas the massa caudalis possesses a coat with galactose as terminal residue; (ii) in RF proper and the massa caudalis the spatial arrangement of glycoproteins might be different. Routine transmission electron-microscopic observations indicate that in larvae of Geotria australis an open communication exists between the ampulla caudalis and blood capillaries via large cavities or lacunae.
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  • 66
    ISSN: 1432-0878
    Keywords: Nasal mucosa ; Odorant-binding protein ; 2-isobutyl-3-methoxypyrazine ; Immunocytochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Polyclonal antibodies have been raised against purified bovine pyrazine-binding protein, a protein that binds the odorant 2-isobutyl-3-methoxypyrazine. These antibodies have been utilized in immunocytochemical experiments to localize the pyrazine-binding protein in bovine nasal mucosa. Tissue fragments, macroscopically identified as olfactory and respiratory mucosa, were fixed in Bouin's fluid and embedded in paraffin. Consecutive serial sections were processed for immunofluorescence studies and restained either with haematoxylin-eosin or with periodic acid Schiff-Alcian Blue. In both olfactory and respiratory mucosa, only seromucous tubulo-acinar glands were specifically labelled. These glands are located in the lamina propria underlying typical respiratory epithelium, even in those tissues that are macroscopically defined as olfactory mucosa.
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  • 67
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Retina ; Immunocytochemistry ; Radioimmunoassay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of neuropeptide Y (NPY)-like immunoreactivity in rat, rabbit, chick, frog and goldfish retinas was investigated by immunohistochemistry. Positive results were observed only in the frog and goldfish retinas. NPY immunoreactivity was associated with a small population of amacrine cell bodies in the inner nuclear layer and cell processes in the inner plexiform layer of both retinas. In the frog retina, three distinct layers containing immunoreactivity were observed in the inner plexiform layer. In contrast, the immunoreactivity in the same area of the goldfish retina was more or less separated into two layers. Convincing evidence could not be found for the co-existence of NPY-like material with other putative transmitter-like substances in the two retinas. Radioimmunoassay revealed the presence of small amounts of NPY-like immunoreactivity in the rabbit retina; the goldfish and frog retinas contained significantly more immunoreactive material. High performance liquid chromatography of the immunoreactive material in frog and goldfish retinas showed each retina containing different molecular forms of NPY-like proteins, neither of which resembled porcine NPY or PYY. The endogenous NPY-like material of the frog retina can be released by potassium depolarisation in a calciumdependent way. In view of all these data an NPY-like protein must now be considered a potential retinal transmitter.
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  • 68
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    Cell & tissue research 248 (1987), S. 393-398 
    ISSN: 1432-0878
    Keywords: Adrenaline ; Phenylethanolamine-N-methyl transferase ; Immunocytochemistry ; Electron microscopy ; Central nervous system ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of adrenergic axon terminals was examined in the paraventricular nucleus of the thalamus (PNT) and in the hypothalamic arcuate nucleus-median eminence (ARC-ME) complex by use of phenylethanolamine-N-methyl transferase (PNMT) immunocytochemistry. In the PNT, immunoreactive terminals formed a dense and well-circumscribed plexus. In the ARC, labeled varicosities were less numerous and more evenly distributed. In the ME, they were scarce and confined to the inner zone. In all these areas, the diameter of immunoreactive varicosities ranged between 0.2 and 1.3 μm; in the ME and in the transitional zone between the ARC and the ME, a population of larger boutons (〉2 μm) was also visible. All immunoreactive varicosities exhibited densely packed small, clear vesicles associated with a few large granular vesicles. In the PNT and the ARC, but not in the ME, they formed synaptic contacts with dendritic elements and were occasionally apposed to neuronal cell bodies. These axo-somatic appositions showed no junctional specializations. In the ME and transitional zone, immunoreactive terminals were frequently juxtaposed to, and occasionally established differentiated synaptic contacts with, tanycytes. These data support a transmitter role for adrenaline in the diencephalon and suggest that adrenaline plays a role in hypothalamo-hypophysiotropic regulation through interactions with neural and glial elements.
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  • 69
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    Cell & tissue research 248 (1987), S. 409-415 
    ISSN: 1432-0878
    Keywords: Bone matrix ; Osteonectin ; Osteoblasts ; Immunocytochemistry ; Differentiation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Bone matrix consists of type-I collagen and noncollagenous proteins. The latter represent only 10% of its total protein content. Since type-I collagen is also present in various other connective tissue sites (e.g., skin) it cannot be considered as bone specific. Among the non-collagenous components osteonectin — a 32 kilodalton (KD) glycoprotein linking mineral to collagen fibrils — is thought to be bone specific due to its biochemical properties. In the present study various skeletal and non-skeletal tissues were investigated for the presence of osteonectin by means of immunocytochemical methods. Two polyclonal antibodies against human and bovine osteonectin were applied. Immunocytochemically, osteonectin could be demonstrated in active osteoblasts and osteoprogenitor cells as well as in young osteocytes, while aged, quiescent osteocytes did not contain the protein, suggesting that the protein is a marker of the osteoblastic functional differentiation of bone cells. Osteonectin was absent in all non-skeletal tissues with the exception of chondrocytes in so-called mineralizing chondroid bone.
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  • 70
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Serotonin ; Diphyllobothrium dendriticum ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the nervous system of the obligatory endoparasite Diphyllobothrium dendriticum immunoreactivity (IR) to growth hormone-releasing factor (GRF), peptide histidine isoleucine (PHI), bovine pancreatic polypeptide (BPP), gastrin, gastrin-releasing peptide (GRP), oxytocin, FMRF-amide (FMRF) and serotonin (5HT) was demonstrated by immunocytochemical methods. A very strong GRF-IR was observed in the CNS and PNS of larvae and of the constantly growing adult worms. GRF-IR axon terminals occur beneath the basal lamina of the tegument along the inside of the bothridia, the holdfast organ of the worm. GRF-IR fibres surround the yolk producing vitelline glands and occur in the wall of the vagina. PHI-IR was observed in the CNS and PNS of larvae and adult worms. PHI-IR terminals occur beneath the basal lamina of the tegument along the strobila, the nutrient absorbing surface of the worm. PHI-IR fibres seem to innervate the testicular follicles. FMRF-IR fibres and perikarya occur close to the vitelline glands and the uterine pore and in the male copulatory organ. Numerous large 5HT-IR perikarya with long varicose fibres were observed in the nervous system of the worm. 5HT-IR perikarya occur close to the genital atrium. D. dendriticum is the phylogenetically lowest organism in which IR to PHI has been demonstrated.
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  • 71
    ISSN: 1432-0878
    Keywords: Adipokinetic hormone ; Corpus cardiacum ; Locusta migratoria ; Periplaneta americana ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunocytochemical method was used to differentiate between immunoreactive substances in glandular cells in the corpora cardiaca (CC) and in certain cerebral neurons in 2 insect species, Locusta migratoria migratorioides and Periplaneta americana. The staining properties of antisera raised to different parts of the decapeptide adipokinetic hormone (AKH) were compared and their specificity was determined by preabsorption with AKH and related peptides. Antibodies raised to the N-terminal part of AKH (serum 433) and the central and C-terminal part (serum 241) were found to have different staining properties. In the CC of the locust both antisera show a strong immunoreactivity with glandular cells, we therefore suggest that at least one of the compounds revealed is AKH. Some of the glandular cells in the locust and large numbers of glandular cells in the CC of the cockroach are revealed by the N-terminal specific antiserum. On the other hand, neurons in the central nervous system are revealed only by the C-terminal specific antiserum. The possible identity of the various substances revealed by these two antisera is discussed.
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  • 72
    ISSN: 1432-0878
    Keywords: Embryonic hypothalamo-hypophysial system ; Vasotocin neurons ; Mesotocin neurons ; Chick embryo hypothalamus ; Immunocytochemistry ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothalamo-hypophysial system of the chick embryo has been studied with a monoclonal antibody which cross-reacts with arginine vasotocin and mesotocin, using thick (100 μm) sections in conjunction with a peroxidase-conjugated rabbit anti-mouse antibody. Although weakly stained perikarya occur occasionally in the tuberal region on embryonic days 6 and 7, the most consistent immunostaining of perikarya is found in the periventricular region of the caudal midhypothalamus at the level of the optic chiasm after embryonic day 8 1/2. Synthesis of peptides, therefore, takes place while the cells are close to their site of origin. Between embryonic days 9 and 10, beaded axons run along the anterior median eminence closely apposed to the adenohypophysis, thereby forming the anlage of the zona externa. The axons of the hypothalamo-neurohypophysial tract surround the neural lobe between embryonic days 11 and 12. The caudal to rostral wave of neuronal maturation that occurs during development appears to be due to a progressive differentiation of the periventricular zone, as well as the migration of perikarya. The early periventricular perikarya at embryonic day 8 1/2 send processes rostrally in a wing-shaped formation that extends both dorso- and ventrolaterally. From embryonic days 10 to 12, perikarya can be observed in the wing-like extensions, apparently migrating to rostral levels. The dorsolateral pathway gives rise at its midportion to the lateral cell group, whereas those perikarya migrating more laterally form the anlage of the external supraoptic nucleus. The ventrolateral wing-shaped extension of perikarya appears to be directed toward the ventral group and those lateral perikarya continuous with it. The location of mature neuronal cell groups is well established by embryonic day 17.
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  • 73
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    Cell & tissue research 243 (1986), S. 33-40 
    ISSN: 1432-0878
    Keywords: Ornithine decarboxylase ; Macrophage ; Immunocytochemistry ; Murine cell culture ; Antibody specificity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antiserum elicited to ornithine decarboxylase (ODC) purified from murine RAW 264 macrophage-like cells has been employed to localize ODC in cultured murine cells. The antiserum immunoprecipitated 100% of the ODC activity from the cultured cells. The specificity of the antiserum was demonstrated by the immunoprecipitation from 35S-methionine metabolically-labeled cell extracts of a single protein which migrated upon SDS-gel electrophoresis coincident with authentic ODC. Indirect immunofluorescence experiments were performed on paraformaldehyde-fixed RAW 264 cells and JB6 epidermal cells using the rabbit anti-ODC antiserum and FITC-conjugated goat anti-rabbit IgG. Little immunofluorescence was apparent in non-stimulated cells. Intense immunofluorescence was detectable in stimulated cells at times of peak cellular ODC activity. Antigenically-reactive ODC was localized diffusely in the cytoplasm and was absent in the nuclei of RAW 264 cells, whereas in the JB6 cells the immunodetectable enzyme protein was localized in a punctate pattern in both the cytoplasm and nucleoplasm and was absent in the nucleolus. The appearance and disappearance of immunoreactive ODC in both cell types after stimulation was consistent with the alterations in ODC activity.
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  • 74
    ISSN: 1432-0878
    Keywords: Insect visual system ; Photoreceptors ; Neurotransmitter ; Histamine ; Immunocytochemistry ; Calliphora erythrocephala ; Musca domestica (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies to histamine were used for immunocytochemical studies of the visual system in the flies Calliphora erythrocephala and Musca domestica. Specific immunolabeling of photoreceptors was found both in the compound eyes and ocelli of both species. In the compound eyes histamine-like immunoreactivity (HA-IR) was found in all the short visual fibers (photoreceptors R1–6) and one type of long visual fiber (photoreceptor R8). In addition, the ocellar photoreceptors also show HA-IR. In view of earlier biochemical and pharmacological/physiological findings by Elias and Evans (1983) and Hardie (1987) it thus seems likely that histamine is a neurotransmitter in insect photoreceptors. Interestingly, the second type of long visual fiber (photoreceptor R7) has recently been found to be GABA-immunoreactive (Datum et al. 1986). The two types of long visual fibers may hence use different transmitters which act on different receptors of the postsynaptic neurons in the second visual neuropil, the medulla. In addition to the photoreceptors in the retina and ocelli, we found processes of HA-IR neurons in one of the optic lobe neuropils, the lobula. This finding indicates that histamine may also be a transmitter in certain interneurons in the visual system.
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  • 75
    ISSN: 1432-0878
    Keywords: Vasopressin precursor ; Endoplasmic reticulum ; Gold/silver intensification ; Immunocytochemistry ; Brattleboro rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This ultrastructural study demonstrates that the vasopressin immunoreactivity found in the occasional, densely stained cells in the hypothalamus of the homozygous Brattleboro rat is localized in the rough endoplasmic reticulum. 50-μm Vibratome sections were stained with anti-vasopressin serum by use of a peroxidase method with 3,3-diaminobenzidine as chromogen. The diaminobenzidine end-product has a specific capability to bind gold particles from a chloroauric acid solution and the bound gold was used to precipitate silver grains from a silver developer. The stained sections were flat embedded in resin and ultrathin sections were cut of areas containing the immuno-identified occasional cells. In these densely stained, vasopressin-immunoreactive cells of homozygous Brattleboro rats the rough endoplasmic reticulum was dilated. The lumen of the reticulum contained both end-products of diaminobenzidine and gold/silver grains, but some parts of the reticulum appeared unstained. No other cell organelles were immunostained and no secretory granules were found. In control rats, gold/silver deposits were found throughout the cytoplasm of vasopressin-immunoreactive cells. In these immunostained cells secretory granules were seen.
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  • 76
    ISSN: 1432-0878
    Keywords: Peripheral neurosecretory structures ; Immunocytochemistry ; Insulin ; Glucagon ; AKH ; Insects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Insulin, glucagon and adipokinetic hormone antisera were applied to the corpora cardiaca, perisympathetic organs, neurohemal areas and peripheral neurosecretory cells of three insect species, the locust Locusta migratoria, the cockroach Periplaneta americana, and the stick insect Carausius morosus. The neurohemal part of the corpora cardiaca was shown to be immunoreactive to both insulin and glucagon antisera while the glandular cells reacted to adipokinetic hormone antisera. The perisympathetic organs seem to be devoid of these three substances, but certain peripheral neurohemal areas contained AKH and glucagon immunoreactive products. The latter were found to originate in the peripheral neurosecretory cells.
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  • 77
    ISSN: 1432-0878
    Keywords: Inner ear ; Cochlea ; Actin ; Immunocytochemistry ; Chinchilla
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cochleas from chinchilla inner ears were processed in the cold through Lowicryl K4M, and cured by UV light. Thick (2 μm) sections were reacted with primary antibodies raised against actin, and anti-actin antibodies localized by FITC epifluorescence. On thin sections from the same blocks anti-actin antibodies were localized ultrastructurally with secondary antibodies coupled to colloidal gold. In the hair cells, actin was present in the stereocilia and cuticular plate, regions where thin filaments were observed by electron microscopy. Colloidal gold was uniformly distributed over these regions and over the stereocilia rootlets demonstrating that actin was present in this region although previously in permeabilized cells, the rootlet was not decorated with myosin subfragment S-1. Actin was present in the pillar and Deiters supporting cells at the reticular lamina and at the basilar membrane, where a meshwork of thin filaments was seen by electron microscopy. Colloidal gold particles were also localized over the thin processes of the pillar and Deiters cells, and over the region of the Deiters cell which envelops the base of the outer hair cell. In these regions actin co-localized with microtubules along the entire length of the supporting cells.
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  • 78
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Glycoproteins ; Secretory process ; Immunocytochemistry ; Lectin histochemistry ; Rat ; Vertebrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO) of the rat was investigated by use of histochemical and immunocytochemical methods at the light and electron-microscopic levels. Consecutive thin methacrylate sections were stained with the pseudoisocyanin (Psi), immunoperoxidase (IMC; employing an antiserum against Reissner's fiber, AFRU), periodic acid-Schiff (PAS) and periodic acid-silver methenamine (SM) techniques, and reacted with six types of lectins. Psi, SM, concanavalin A (Con A) and IMC were also used for double and triple sequential staining of the same section. Increasing dilutions of AFRU (from 1∶1000 to 1∶200 000) were used for immunostaining of serial paraffin sections. In addition, ultrastructural localization of (i) Con A-binding sites and (ii) immunoreactive secretory material was performed. Some of these procedures were also applied to the ophidian and canine SCO. Con A-positive, Psi-positive and immunoreactive materials coexisted within the same cisternae of the rough endoplasmic reticulum. The Golgi apparatus lacked Con A-positive and immunoreactive substances. Apical secretory granules and secreted material lying on the surface of the SCO showed (i) the highest affinity for AFRU, but were (ii) Con A-negative, and (iii) wheat-germ agglutinin-, PAS and SM-positive. Reissner's fiber displayed a low affinity for AFRU. It is suggested that the SCO secretes N-linked glycoproteins, the carbohydrate and protein moeities of which undergo (i) a maturation process before being released, and (ii) some kind of modification(s) after their release into the ventricle. The perivascular secretory cells of the dog SCO might secrete a material different from that secreted by the ependymal cells.
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  • 79
    ISSN: 1432-0878
    Keywords: Crustacean cardioactive peptide (CCAP) ; Proctolin ; FMRFamide ; Leu-enkephalin ; Immunocytochemistry ; Ultrastructural immunogold-labeling ; Pericardial organs ; Neurosecretion ; Carcinus maenas (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Polyclonal antibodies were raised in rabbits against synthetic crustacean cardioactive peptide (CCAP) conjugated to bovine thyroglobulin, and were used to map CCAP-immunoreactive structures in the central nervous system of Carcinus maenas. As expected, the neurohemal pericardial organs (PO) displayed abundant immunoreactivity in nerve fibers and terminals. In addition, immunoreactive neurons were demonstrated in other parts of the nervous system. At least some of them do not appear to terminate in neurohemal structures and may have a non-endocrine, as yet unknown function. Immunoreactive perikarya with a diameter of 25–30 μm occur in the brain. They project into the optic and antennary neuropil, and into the eyestalk. One cell was found in the medulla terminalis of the eyestalk and in the connective ganglion, respectively. From the latter, axonal branches could be traced into the brain and the thoracic ganglia (TG). In the TG, small-diameter perikarya give rise to extensive networks of varicose fibers. Some of the perikarya occur in a characteristic paired arrangement with larger CCAP-immunoreactive somata (diameter 40–50 μm). These pairs of one small and one large cell occur in all mouthpart and leg segments of the TG, except the abdominal ganglia (AG), where only large cells were found. The main projections of the large neurons comprise one or more fibers in each of the seven segmental nerves (SN), leading to neurosecretory terminals in the PO. The fibers in the SN are joined by branches of an ascending axonal tract from the large perikarya in the AG. The large-type perikarya are considered to be the principal source of CCAP in the PO. The optic ganglia in the eyestalk, except the medulla terminalis, the neurohemal sinus gland and the stomatogastric nervous system are devoid of CCAP-immunoreactivity. In axon terminals of the PO, CCAP is not colocalized with other PO-neuropeptides, i.e. proctolin-, FMRFamide-like, and Leu-enkephalin-like immunoreactive materials. Electron-microscopic immunocytochemistry revealed a distinct CCAP-containing granule type in specific axon profiles and terminals in the PO. The architecture of CCAP-immunoreactive neurons is discussed with respect to previous morphological studies on the origin and pathways of fibers terminating in the PO.
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  • 80
    ISSN: 1432-0878
    Keywords: Growth hormone ; Prolactin ; Somatostatin ; Immunocytochemistry ; Lamprey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Growth hormone, prolactin and somatostatinlike immunoreactivities were demonstrated in the brains of larval, young adult (parasitic) and upstream migrant adult sea lampreys, Petromyzon marinus, by means of immunoperoxidase techniques. Growth hormone (GH) and prolactin (PRL) were observed within separate perikarya in the nucleus praeopticus, within fibers in the commissura praeinfundibularis, and in nerve endings within the neurohypophysis of larval and adult-stage lampreys. Cell bodies demonstrating immunoreactive growth hormone were more numerous than those reactive for prolactin. Unlike in the upstream migrant adult lamprey, no GH or PRL was demonstrated in the adenohypophysis of larval or parasitic lamprey. Somatostatin (SRIF)-like immunoreactive neurons were demonstrated in the nucleus commissurae praeinfundibularis, anterior and posterior pars ventralis hypothalami, pars dorsalis thalami, and the tegmentum motorium rhombencephali of larval, parasitic and upstream migrant adult lampreys. Many of the SRIF containing neurons within the hypothalamus were cerebrospinal fluid (CSF)-contacting cells. SRIF fibers were found throughout most of the brain predominating within the nucleus praeopticus, pars ventralis hypothalami, and the nucleus interpeduncularis. No SRIF immunoreactivity was found within the neurophyophysis. The possible functions of these peptides within the brain of the lamprey are discussed.
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  • 81
    ISSN: 1432-0878
    Keywords: Insect brain ; Neurotransmitters ; Immunocytochemistry ; Drosophila melanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a monoclonal antibody selective for the acetylcholine (ACh)-synthesizing enzyme choline acetyltransferase (ChAT) of Drosophila melanogaster we find ChAT-like immunoreactivity in specific synaptic regions throughout the brain of Drosophila melanogaster apart from the lobes and the peduncle of the mushroom body and most of the first visual neuropile (lamina). Several anatomically well-defined central brain structures exhibit particularly strong binding. Characteristic differential staining patterns are observed for each of the four neuromeres of the optic lobes. Cell bodies appear not to bind this antibody. The prominent features of the distribution of ChAT-like immunoreactivity are paralleled by the distribution of acetylcholine hydrolyzing enzymatic activity as revealed by histochemical staining for acetylcholine esterase (AChE). These results are discussed in comparison with published data on enzyme distribution, choline uptake and ACh receptor binding in the nervous system of Drosophila melanogaster.
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  • 82
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    Cell & tissue research 244 (1986), S. 549-555 
    ISSN: 1432-0878
    Keywords: Fibronectin ; Lamprey ; Hagfish ; Immunocytochemistry ; Connective tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactive fibronectin-like material was localized within tissues of agnathans (hagfishes and lampreys) by an immunoperoxidase technique. Fibronectin was detected in basement membranes and in loose and dense connective tissues throughout the agnathan body. A fibronectin-like component was also identified in the plasma of both lampreys and hagfishes. The results indicate that fibronectin or a fibronectin-like material is a major component of agnathan connective tissues. Although there were some variations in the localization of fibronectin both between the lamprey and the hagfish and between agnathan and other vertebrate tissues, the generalized pattern of distribution of fibronectin in the agnathans supports the view that this protein, like that in higher vertebrates, plays a role in cellmatrix adhesion and tissue organization.
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  • 83
    ISSN: 1432-0878
    Keywords: Human growth hormone-releasing factor (hGRF) ; Paraventricular nucleus ; Immunocytochemistry ; Electron microscopy ; Synapses ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of a preembedding immuno-electronmicroscopic technique, a large number of nerve endings containing a substance related to human growth hormonereleasing factor (hGRF) have been demonstrated in the paraventricular nucleus of the guinea pig. They made synaptic contacts primarily with dendritic shafts: 80% of these contacts were symmetrical. The immunoprecipitate was located mainly in large granules and around small clear vesicles. These findings suggest that a peptide related to hGRF may play a role in neural communication in the paraventricular nucleus.
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  • 84
    ISSN: 1432-0878
    Keywords: Thyrotropin-releasing hormone (TRH) ; Development, ontogenetic ; Anterior hypothalamus ; Immunocytochemistry ; Domestic mallard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Developmental changes of thyrotropin-releasing hormone (TRH)-immunoreactive structures in the brain of mallard embryos were studied by means of immunocytochemistry (PAP technique). The primary antibody was generated against synthetic TRH. Immunoreactive neurons were first detected in the hypothalamus of 14-day-old embryos. By day 20, increasing numbers of immunoreactive perikarya were observed in the paraventricular nucleus, anterior preoptic region and supraoptic region. Immunoreactive fiber projections were seen in the median eminence as early as embryonic day 20; they occurred also in some extrahypothalamic regions (lateral septum, accumbens nucleus). The number and staining intensity of the cell bodies increased up to hatching, and continued to increase during the first week after hatching.
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  • 85
    ISSN: 1432-0878
    Keywords: Pancreas ; Exocrine glands ; Islets of Langerhans ; Differentiation ; Immunocytochemistry ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The islet cells of the mammalian pancreas are comprised of four different endocrine cell types, each containing a specific hormone. Islet cells also contain two enzymes of the catecholamine biosynthetic pathway: tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC). The cell lineage relationships of these different cell types have not been examined and it is not known whether, during development, they originate from the same or from different precursor populations. In this study we used immunocytochemical procedures to determine whether developing pancreatic cells express markers common to endocrine and exocrine cell types. We found that acinar cell precursors express AADC prior to the appearance of an exocrine marker and that the expression of AADC in acinar cells persists throughout embryogenesis to the first month of postnatal life. At this time, acinar cells do not contain AADC. We also found that exocrine cells containing AADC never express other islet-cell markers. These findings suggest that while acinar and islet cells both arise from precursor cells containing AADC, these progenitor cells do not express a combined endocrine-exocrine phenotype.
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  • 86
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Alkaline phosphatase ; Small intestine ; Colchicine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The purpose of this study was to investigate the effect of colchicine and vinblastine on the localization of alkaline phosphatase (AlPase) in rat duodenum in relation to structural changes. AlPase was localized on the membranes of rough endoplasmic reticulum, Golgi stacks, cytoplasmic vesicles, microvilli, on lateral plasma membranes, and in some lysosomes of the duodenal epithelial cells of rats treated with either lumicolchicine or 0.9% NaCl alone. Microvilli were most intensely stained, and AlPase-positive Golgi stacks were regularly distributed in the supranuclear regions. After colchicine treatment, microvilli were shortened and the staining intensity became weaker, whereas basal as well as lateral plasma membranes showed stronger staining. The AlPase-positive microvilli appeared not only on the luminal surfaces, but also on the baso-lateral plasma membranes and even on the surfaces of characteristic intracytoplasmic cysts. Golgi stacks became smaller and their distribution became less localized, and the staining intensity of the Golgi stacks became weaker. AlPase localization in rats treated with vinblastine was almost identical with that of rats treated with colchicine. Thus, colchicine and vinblastine appeared to have elicited a disorientation of intracellular transport of intestinal AlPase by inhibiting microtubule organization.
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  • 87
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    Cell & tissue research 245 (1986), S. 457-460 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Somatotroph ; Lactotroph ; Ultrastructure ; Immunocytochemistry ; Fetal and neonatal rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of immunoreactive somatotrophs and lactotrophs in pituitaries of fetal rats at 19, 20 and 21 days of gestation and on the day of birth was studied. Somatotrophs, first detectable at 19 days of gestation, undergo only minor modifications before reaching the structure described for adults. In particular there is an increase in the endoplasmic reticulum and Golgi apparatus. Lactotrophs, first identifiable in newborn rats, are very different in ultrastructure from adult cells, because the secretory granules are generally small, but variable in shape and size, and the Golgi complex is prominent.
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  • 88
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone ; Ultrastructural histochemistry ; Immunocytochemistry ; Neurosecretion ; Crustaceans (Carcinus maenas, Cancer pagurus, Uca pugilator, Orconectes limosus)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A high-liter antiserum has been obtained from two rabbits immunized with a glutaraldehyde conjugate of synthetic pigment-dispersing hormone (PDH) from Uca pugilator and bovine thyroglobulin. The antiserum blocked melanophore-dispersing activity of the peptide in vivo. In sinus glands (SG) of Carcinus maenas, Cancer pagurus, Uca pugilator and Orconectes limosus, electron-microscopic immunocytochemistry revealed sparsely distributed axon endings containing a distinct PDH-immunoreactive type of neurosecretory granules (diameter 90–130 nm). Exocytotic figures indicating release of the content of these granules into hemolymph lacunae were occasionally observed. Preservation of fine structure and antigenicity of the PDH granules were markedly dependent on the fixation procedure used. A preliminary experiment with C. maenas showed that preterminal axon dilatations near the basal lamina seemed to accumulate PDH-granules when animals were kept in complete darkness for three days. Immunodot blotting of fractions after high pressure liquid chromatography (HPLC) of extracts from SGs of C. maenas and O. limosus revealed a strongly immunoreactive substance at a retention time very similar to those of synthetic PDHs of Uca pugilator and Pandalus borealis. It is also coincident with a zone of biological activity. Thus, the antigen demonstrated by immunocytochemistry is identical or very similar to one of the known PDHs.
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  • 89
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone (crustaceans) ; FMRFamide ; Immunocytochemistry ; Neurosecretion ; Crustaceans: Carcinus maenas, Orconectes limosus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of a new antiserum, raised against synthetic pigment-dispersing hormone (PDH) from Uca pugilator, immunoreactive structures were studied at the light-microscopic level in the eyestalk ganglia of Carcinus maenas and Orconectes limosus. PDH-reactivity was mainly found in two types of neurons that were located between the medulla interna (MI) and the medulla terminalis (MT) in both species. Several additional perikarya were located in the distal part of the MI in O. limosus. In C. maenas, two to three PDH-positive perikarya were found in the region of the X-organ (XO) in the MT. Processes from single and clustered cells could be traced into all medullae of the eyestalk. Axons from the immunoreactive perikarya running between MI and MT form a larger tract that traverses the MT. Fibers from this tract give rise to extensive arborizations and plexuses throughout the proximal MT. A plexus containing very fine fibers is located at the surface of the MT in a position distal to the XO-area of C. maenas only. The proximal plexus also receives PDH-positive fibers through the optic nerve. PDH-perikarya in the cerebral ganglion may also project into the more distal regions of the eyestalk. Distal projections of the perikarya between the MI and MT consist of several branches. Most of these are directed toward the MI and ME (medulla externa) wherein they form highly organized, layered plexuses. One branch was traced into the principal neurohemal organ, the sinus gland (SG). In the SG, the tract gives off arborizations and neurosecretory terminals. It then proceeds in a proximal direction out of the SG, adjacent to the MT. Its further course could not be elucidated. The lamina ganglionaris (LG) receives PDH-fibers from the ME and fine processes from small perikarya located in close association with the LG in the distal part of the first optic chiasma. The architecture of PDH-positive elements was similar in both C. maenas and O. limosus. The distribution of these structures suggests that PDH is not only a neurohormone but may, in addition, have a role as a neurotransmitter or modulator. Immunostaining of successive sections with an FMRF-amide antiserum revealed co-localization of FMRFamideand PDH-immunoreactivities in most, but not all PDH-containing perikarya and fibers. The axonal branch leading to the SG and the SG proper were devoid of FMRFamide immunoreactivity.
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  • 90
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    Cell & tissue research 251 (1988), S. 233-236 
    ISSN: 1432-0878
    Keywords: Prolactin ; Corpus luteum ; Immunocytochemistry ; Luteal cells ; Luteotropic function ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Six human corpora lutea (day 17–25) of the menstrual cycle and 4 ovarian stromal tissues from 7 cycling women were examined for the presence of the hormone, prolactin, by immunohistochemistry using the indirect peroxidase-antiperoxidase method. After mounting tissue sections of 4 μm, endogenous peroxidases were removed with hydrogen peroxide and the sections were incubated for l h at room temperature followed by 16 h at 4° C with a highly specific antisera for human prolactin, nonimmunized normal rabbit serum for a control reaction, or antiserum preadsorbed with excess human prolactin for specificity determination. Following the reaction with the second antibody (goat antirabbit IgG) for l h at room temperature, prolactin was localized using peroxidase anti-peroxidase and 3.3′-diaminobenzidine as the chromogen. Prolactin was present and could be localized in the luteal cells of all 6 corpora lutea, but not in any of the ovarian stroma studied. Human adenohypophysis served as a positive tissue control for prolactin immunopositive staining. The localization of immunoreactive prolactin in the corpus luteum demonstrates directly the presence of this hormone in the human ovary, adding further evidence for its role in luteal function.
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  • 91
    ISSN: 1432-0878
    Keywords: Growth ; Enkephalin ; Endogenous opioids ; Immunocytochemistry ; Differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of enkephalin, an endogenous opioid, in tissues and cells of the developing and adult rat was determined by immunocytochemistry with antibodies to met- and leu-enkephalin. Met- and leu-enkephalin were found in all developing cells investigated, with staining generally located throughout the cytoplasm; cell nuclei were not immunoreactive. In comparison to developing cells, immunoreactive analogues to met-enkephalin were usually difficult to detect in the adult. Some notable exceptions were reaction products in leukocytes in blood, lung, and cortex of thymus, fibroblasts in the skin, and seminiferous tubules. These results, in concert with earlier reports that opioid receptors are found largely in developing, but not adult, tissues, indicate that endogenous opioids are specifically involved in biological development, particularly cell proliferation and differentiation. Immunoreactivity in adult nonneural cells may be related to their development in some cases, but also could indicate other functions.
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  • 92
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    Cell & tissue research 252 (1988), S. 9-15 
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Insulin ; Immunocytochemistry ; Lysosomes ; Crinophagy ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural studies of pancreatic islets have suggested that crinophagy provides a possible mechanism for intracellular degradation of insulin in the insulin-producing B-cells. In the present study, a quantitative estimation of crinophagy in mouse pancreatic islets was attempted by morphometric analysis of lysosomes containing immunoreactive insulin. Isolated islets were incubated in tissue culture for one week in 3.3, 5.5 or 28 mmol/l glucose. The lysosomes of the pancreatic B-cells were identified by morphological and enzyme-cytochemical criteria and divided into three subpopulations comprising primary lysosomes and insulin-positive or insulin-negative secondary lysosomes. Both the volume and numerical density of the primary lysosomes increased with increasing glucose concentration. The proportion of insulin-containing secondary lysosomes was highest at 5.5 and lowest at 3.3 mmol/l glucose. Insulin-negative secondary lysosomes predominated at 3.3 mmol/l glucose. Studies of the dose-response relationships of glucose-stimulated insulin biosynthesis and insulin secretion of the pancreatic islets showed that biosynthesis had an apparent Km-value for glucose of 7.0 mmol/l, whereas it was 14.5 mmol/l for secretion. The pronounced crinophagic activity at 5.5 mmol/l glucose may thus be explained by the difference in glucose sensitivity between insulin biosynthesis and secretion resulting in an intracellular accumulation of insulin-containing secretory granules. The predominance of insulin-negative secondary lysosomes at 3.3 mmol/l glucose may reflect an increased autophagy, whereas the predominance of primary lysosomes at 28 mmol/l glucose may reflect a generally low activity of intracellular degradative processes.
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  • 93
    ISSN: 1432-0878
    Keywords: Spermatozoa ; Seminal vesicles ; Seminal fluid ; Sperm motility ; Immunocytochemistry ; Bull
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The seminal vesicles synthesize in an androgen-dependent manner a neutral protein of 13.5 kDa molecular weight that makes up about 40% of their secretion (“major protein”). An antiserum against this protein raised in rabbits was used to localize the antigen within the seminal vesicles. In addition to intraluminal secretion of the seminal vesicles and the ampulla of the vas deferens, ejaculated and ampullary spermatozoa revealed an intense immunoreaction, which was restricted to the neck region of the sperm head and the middle piece, while the principal piece of the tail as well as the sperm head were devoid of immunoreactive material. Comparison of spermatozoa taken from the tail of the epididymis with ampullary spermatozoa showed that about 90% of the latter, but only 10–20% of the former presented this distributional pattern of immunoreactive sites. Epididymal epithelium as well as calf seminal vesicle epithelium showed no immunoreactivity with major protein antiserum. Using a pre-embedding staining technique with gold-labeled primary or secondary antibodies, respectively, no immunostaining could be achieved at the ultrastructural level. Incubation experiments of epididymal spermatozoa in EGTA-containing solutions in the absence of calcium resulted in a gradual labilization and eventual loss of the plasma membrane of the sperm middle piece. After removal of (at least part of) the plasma membrane, bound major protein could be visualized immunohistochemically close to the mitochondria of the middle piece using a gold-labeled primary or secondary antibody. The acceptor site for major protein therefore seems to reside inside the plasma membrane of the sperm middle piece. Incubation of epididymal spermatozoa in phospholipase-containing solutions removed the acceptor site from the spermatozoa. Separation by polyacrylamide treatment of proteins from epididymal sperm cells extracted by sodium hydroxide or phospholipase treatment, subsequently transblotted on nitrocellulose sheets and directly labeled with gold-tagged major protein, demonstrated a protein duplet with a molecular weight of 65 and 67 kDa, respectively, which appears to represent the specific binder of major protein underneath the sperm surface. Binding of major protein to this ∼66 kDa acceptor site is regarded as a physiological event that may be related to the onset of hyperactivated sperm motility.
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  • 94
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    Cell & tissue research 253 (1988), S. 137-143 
    ISSN: 1432-0878
    Keywords: Endostyle ; Ultrastructure ; Immunocytochemistry ; 5-Hydroxytryptamine ; Granules ; Ciona intestinalis ; Corella parallelogramma, (Tunicata) ; Ascidia mentula (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular and subcellular distribution of serotonin (5-hydroxytryptamine, 5-HT) in the endostyle of three species of ascidians, Ciona intestinalis, Corella parallelogramma, Ascidia mentula, was studied by light-(immunoperoxidase) and electron-microscopic (immunogold) immunocytochemistry. At the light-microscopic level 5-HT-like immunoreactivity (5-HT-LI) was exclusively found in cells located in the lateral portion of the endostyle, between zone 7, known to have iodinating capacity, and zone 8, which consists of ciliated cells. At the electron-microscopic level, the 5-HT-immunoreactive cells were found to correspond to cells containing polymorphous, dense granules, 100–300 nm in diameter. The granules were located in the supranuclear cytoplasm facing the endostyle lumen as well as in the infranuclear cytoplasm facing the extracellular space. Quantification showed that the 5-HT-LI was considerably higher (13–67 times) in cytoplasmic areas containing granules as compared to areas devoid of granules. Most, but not all, of the 5-HT-LI was associated with the dense core of the granules. In conclusion, serotonin-containing cells are located in the peripheral portion of the endostyle, between zones 7 and 8. Serotonin is stored in cytoplasmic granules that are present both in the apical and basal cytoplasm. This suggests the possibility that the cells are bipolar and secrete serotonin both in a basal direction to the extracellular space, and in an apical direction to the pharyngeal lumen.
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  • 95
    ISSN: 1432-0878
    Keywords: Caudal neurosecretory system ; Urotensin ; Methionine-enkephalin ; Immunocytochemistry ; Carp, Cyprinus carpio (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Methionine-enkephalin (Met-enk) was detected by immunocytochemistry and radioimmunoassay in the caudal neuro-secretory system of the carp Cyprinus carpio. Some cells showing urotensin I (UI)-immunoreactivity reacted to Met-enk antiserum, but others did not. Neurons with urotensin II (UII)-immunoreactivity did not react to Met-enk antiserum; neurons with both UI and UII immunoreactivities also displayed a negative Met-enk reaction. Met-enk was detected by radioimmunoassay in the urophysis, although the content was relatively small compared with that found in other parts of the central nervous system and in the pituitary.
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  • 96
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    Cell & tissue research 253 (1988), S. 489-491 
    ISSN: 1432-0878
    Keywords: Nervous system, insects ; Immunocytochemistry ; FMRFamide ; Bovine pancreatic polypeptide ; Locust, Schistocerca gregaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of FMRFamide-like immunoreactivity in the metathoracic ganglion of the locust, Schistocerca gregaria, has been investigated in serial semithin transverse sections with the use of the peroxidase-antiperoxidase (PAP) technique. The topographical distribution of approximately 120 immunopositive neurons was established. Antiserum against bovine pancreatic polypeptide (BPP) stains the same ganglionic cells as FMRFamide-antiserum, yet this staining is largely blocked after preabsorption to FMRFamide. A comparison of these results with those from other studies suggests that there may be more than one type of endogenous RFamide-like peptide.
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  • 97
    ISSN: 1432-0878
    Keywords: Development ; Intrapulmonary chemoreceptor ; Immunocytochemistry ; Morphometry ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution, frequency and size of neuroepithelial bodies (NEB) were studied in lungs of rabbits during different stages of development (27-day fetus, newborn, 6, 11, 21, 28 and 56 days postnatally). NEB were visualized by immunostaining with monoclonal antibody against serotonin. Detailed quantitiation of NEB was performed by use of camera lucida drawings of immunostained serial sections from the same anatomical region, i.e. the lower lobe of the left lung. The total number of NEB was counted and expressed per epithelial length of airway, surface area and volume. The size of NEB defined as surface area as well as the position of NEB in relation to the airway bifurcations was assessed in airways of different sizes. The overall number and size of NEB were found to increase during the immediate perinatal period followed by a sharp decline at 56 days of age. The number of NEB peaked at 6 days postnatally (mean 175.5 NEB/mm3 of airway epithelium) and declined significantly (3.0 NEB/mm3) at 56 days of postnatal age. The size of NEB reached its maximum at 11 days (mean surface area 659.54 μm2, with the largest NEB measuring 1839.98 μm2). By 56 days of age, NEB became significantly smaller (mean surface area 177.29 μm2) consisting of small clusters of cells situated deep within the airway epithelium. At all ages, about half of all NEB (mean 47.6%) were localized within the small peripheral airways with up to 63.9% located at airway bifurcations. These findings indicate that the “functional activity” of NEB may be confined predominantly to the perinatal period. The postulated functions of NEB include those of intrapulmonary hypoxia-sensitive chemoreceptors and/or endocrine-paracrine activity in the lung. Such function(s) may be important during adaptation to extrauterine life as well as for growth and development of the lung.
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  • 98
    ISSN: 1432-0878
    Keywords: Peptides ; Small intensely fluorescent (SIF) cell ; Neuropeptide coexistence ; Inferior mesenteric ganglion ; Immunocytochemistry ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Coexistence of peptides in the small intensely fluorescent cells was demonstrated by immunocytochemistry for met-enkephalin-Arg-Gly-Leu, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y and dynorphin. In the extreme example, a single cell was immunoreactive to all 5 peptides examined. Four peptides coexisted in 8% and three peptides in 13% of SIF cells. In 10% of SIF cells no peptide immunoreactivity could be detected. The most prevalent peptide was met-enkephalin (in 46% of cells), then vasoactive intestinal polypeptide (45%), somatostatin (39%), neuropeptide Y (31%) and dynorphin (24%). Met-enkephalin and vasoactive intestinal polypeptide coexisted most commonly (25%).
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  • 99
    ISSN: 1432-0878
    Keywords: L1-antigen ; Cell adhesion molecule ; Developing vestibular neuroepithelium ; Immunocytochemistry ; Mouse (CBAxC57)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Doubts exist as to whether afferent nerve fibers exert a neurotrophic effect on the differentiation of sensory cells in the developing vestibular neuroepithelium. To determine whether innervation of hair cells precedes their differentiation, we have used the L1 adhesion molecule as a marker for axons. The detection of L1 on afferent axons in the otic vesicle of mouse embryos on gestation day 11 shows that nerve fibers penetrate the neuroepithelium before the sensory cells differentiate. L1-immunoreactivity of nerve endings also reveals the considerable fiber ramification on gestation days 14 and 15, i.e., corresponding to the first stages of sensory cell differentiation. The expression of L1 at successive stages of nerve fiber growth in the neuroepithelium, such as fasciculation and ramification, is not consistent with the previous role proposed for L1 as a fascicule-promoting factor and raises the possibility that other mechanisms are involved in L1 mediaded adhesion.
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  • 100
    ISSN: 1432-0878
    Keywords: Hypothalamus ; LHRH-containing neurons ; Chicken LHRH ; Chicken GnRH-II ; Immunocytochemistry ; Domestic fowl ; Japanese quail
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of LHRH-containing perikarya and nerve fibers in the hypothalami of the domestic fowl and Japanese quail was investigated by means of the specific immunoperoxidase ABC method, using antisera against chicken LHRH-I ([Gln8]-LHRH), chicken GnRH-II ([His5-Trp7-Tyr8]-LHRH [2–10]) and mammalian LHRH ([Arg8]-LHRH). Chicken LHRH-I-immunoreactive perikarya were sparsely scattered in the nucleus preopticus periventricularis (POP), nucleus filiformis (FIL) and nucleus septalis medialis (SM), and in bilateral bands extending from these nuclei into the septal area in both species. A few reactive perikarya were also observed in the nucleus accumbens (Ac) and lobus parolfactorius (LPO). Numerous cLHRH-I-immunoreactive fibers were widely scattered in the preoptic, septal and tuberal areas, and were densely concentrated in the external layer of the median eminence and in organum vasculosum of the lamina terminalis (OVLT) in both species. Anti-mammalian LHRH serum cross-reacted weakly with perikarya and fibers immunoreactive to anti-cLHRH-I serum in normal chicken and quail. Anti-cGnRH-II[2–10] serum immunoreacted with magnocellular neurons distributed in the rostral end of the mesencephalon along the midline close to the nervus oculomotorius (N III). These perikarya were apparently different from cLHRH-I immunoreactive neurons. No immunoreactive cells and fibers against anti-cGnRH-II[2–10] were observed in the hypothalamus and median eminence of the chicken or quail. Anti-cGnRH-II[2–10] bound specifically with cGnRH-II. The morphological evidence suggests that cGnRH-II may not be secreted into the portal circulation to act as hypothalamic hormone.
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