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  • Immunocytochemistry  (95)
  • Springer  (95)
  • Cambridge University Press
  • Elsevier
  • Frontiers Media
  • 2020-2022
  • 1985-1989  (78)
  • 1975-1979  (17)
  • 1970-1974
  • 1988  (42)
  • 1985  (36)
  • 1982
  • 1978  (17)
  • 1970
Collection
Publisher
  • Springer  (95)
  • Cambridge University Press
  • Elsevier
  • Frontiers Media
  • Wiley-Blackwell  (2)
Years
  • 2020-2022
  • 1985-1989  (78)
  • 1975-1979  (17)
  • 1970-1974
  • 1980-1984  (29)
Year
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 3
    ISSN: 1432-2048
    Keywords: Avena ; Immunocytochemistry ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytochrome of oat (Avena sativa L., cv. Garry) coleoptile cells in the red-light-absorbing form, Pr, is diffusely distributed while after conversion to the far-red-light-absorbing form, Pfr, it is observed only in very small areas within the cell. Comparison of phytochrome photoversibility measurements to the distribution of the pigment within the cell indicates that the spectral assay is not influenced by the observed compartmentalization of the chromoprotein. However, the observed compartmentalization of phytochrome is correlated with a loss in spectrophotometrically detectable Pr.
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  • 4
    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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  • 5
    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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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    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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  • 7
    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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  • 8
    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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  • 9
    ISSN: 1432-0878
    Keywords: Pineal organ (mammals) ; Retinal S-antigen ; Photoreceptors ; Retina ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of immunocytochemistry retinal S-antigen is selectively demonstrated in retinal photoreceptor cells of the rat and in pinealocytes of the hedgehog, rat, gerbil and cat. Brain areas surrounding the pineal organ are immunonegative. The immunoreactive material is evenly distributed in the perikarya of the cells. Occasionally, inner segments of retinal photoreceptors and processes of pinealocytes are also stained. The outer segments of retinal photoreceptors display a strong immunoreaction. In both pinealocytes and retinal photoreceptors the intensity of the immunoreaction varied considerably among individual cells. The immunocytochemical demonstration of retinal S-antigen in mammalian pinealocytes indicates that these cells still bear characteristics of photoreceptors. This finding is in accord with the concept that mammalian pinealocytes are derived from pineal photoreceptor cells of poikilothermic vertebrates.
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  • 10
    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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  • 11
    ISSN: 1432-0878
    Keywords: Mammalian-type lutropin ; Pituitary ; Immunocytochemistry ; Light microscopy ; Lepidosiren (SouthAmerican lungfish)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of lutropin (LH)-like material in the pituitary gland of the South-American lungfish, Lepidosiren paradoxa, has been demonstrated by means of the unlabeled antibody-enzyme method, by use of rabbit antiovine LH as first antibody. The LH-like substance was revealed in a single PAS-positive cell type primarily located in the anterior part of the distal lobe. Dot blot tests as well as conventional liquid-phase absorption experiments indicate that the anti-ovine LH antibodies possess specificity against the oLH/gb subunit. These observations indicate that dipnoans (Lepidosiren) share a number of antigenic determinants with those of mammalian LH/gb and support the concept that mammalian LH/gb, or part of it, was established early in evolution. The exact nature and physiological function of the substance detected remains to be defined.
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  • 12
    ISSN: 1432-0878
    Keywords: Proctolin ; Neuropeptide ; Immunocytochemistry ; Terminal ganglion ; Periplaneta americana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurons with proctolin-like immunoreactivity were mapped in the terminal ganglion of Periplaneta americana. The effect of different fixation methods on the variability of immunostaining is described and discussed. The appearance of immunoreactive presynaptic terminals, described here for the first time in insects, points to a function of proctolin as neurotransmitter or neuromodulator in the central nervous system of P. americana besides its known role in the periphery. Proctolin-like immunoreactivity was shown in pre- and postsynaptic profiles. Synaptic contacts are described in detail.
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  • 13
    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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  • 14
    ISSN: 1432-0878
    Keywords: Enkephalin-like immunoreactivity ; Paraganglia, human ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of enkephalin-like immunoreactivity in paraganglia of the urinary bladder of adult humans was studied by use of immuno-electron microscopy. All paraganglionic cells were positively stained. Enkephalin-like immunoreactivity was located in chromaffin granules. Chromaffin cells in the paraganglia showed only few degenerative features, suggesting undisturbed function of the cells.
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  • 15
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    Springer
    Cell & tissue research 186 (1978), S. 399-412 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Dog ; Pars distalis ; Thyrotropin (TSH) ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the immunoperoxidase technique and antisera to the specific beta (β) subunits of bovine and rat TSH1, selective immunocytochemical staining was localized in a specific cell population in the pars distalis of the dog pituitary gland. These TSH cells were found to be positive to aldehyde fuchsin, alcian blue, periodic acid-Schiff (PAS) and aniline blue. With the performic acidalcian blue (pH 0.2) -PAS-orange G procedure these cells stained blue-purple, demonstrating FSH/LH cells (blue or turquoise), ACTH/MSH cells (redpurple) and PRL cells (orange-red). The TSH cells were further differentiated from other functional cell types of the pars distalis on the basis of their typical cytological features, intraglandular distribution and by immunocytochemical double staining. In the pars distalis of adult male dogs the TSH cells were mostly shown to be smaller in size and less numerous than in bitches in the anestrous phase of the sexual cycle. Moreover, cytological alterations in the immunoreactive thyrotrophs in the pituitary of male and female dogs generally paralleled the spontaneous changes in thyroid function associated with thyroid atrophy and/or pituitary insufficiency, and thyroid hyperplasia or goiter. In conclusion, because of their specificity and high potency, the antisera to the β-subunits of bovine and rat TSH represent an effective tool for the selective immunocytochemical localization of TSH in the dog pituitary. This allows the study of the morphology and function of TSH cells under different physiological, pathological and experimental conditions.
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  • 16
    Electronic Resource
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    Springer
    Cell & tissue research 188 (1978), S. 99-106 
    ISSN: 1432-0878
    Keywords: LHRH-neurosecretion ; Avian hypothalamus ; Vasotocin neurosecretion ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A fluorescent technique applying specific LHRH and vasotocin antisera was used for the immunocytochemical localization of the respective neurosecretory systems in the hypothalamus of gonadectomized, testosteronetreated and/or serotonin injected male domestic ducks. An immunoreactive (IR) LHRH-producing system, with perikarya located in the preoptic nucleus, could be traced through the ventral hypothalamus down to the external layer of the rostral and caudal ME, in close vicinity to the hypophysial portal system. An IR-vasotocin system originating in the paraventricular and supraoptic nuclei ran through the ventral hypothalamus, but terminated in (i) the external layer of the rostral ME, and (ii) in the posterior lobe of the hypophysis.
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  • 17
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    Cell & tissue research 188 (1978), S. 119-132 
    ISSN: 1432-0878
    Keywords: Neurophysin ; Paraventricular nucleus ; Supraoptic nucleus ; Sheep ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum cross-reactive against ovine neurophysins-I-II and -III has been used in conjunction with the immunoperoxidase histochemical procedure to localize the cells of the sheep paraventricular (PVN) and supraoptic nuclei (SON). In order to describe the topographical distribution of the SON and PVN a study was made on the serial sections cut (a) transversely from rostral to caudal positions and (b) sagittally from lateral to medial positions of the hypothalamus. The cells of the SON, when examined in the transverse aspect, extended approximately 1900 μ caudally and when examined in the sagittal plane were contained within a lateral-medial distance of 4830 μ. In each case the SON cells lay adjacent to the optic chiasm. As sections were cut transversely, the cells of the PVN first appeared in a rostral position defined as 0 μ and close to the ventral lining of the third ventricle. This general ventral and ventro-lateral distribution of cells maintained up to a caudal distance of approximately 840 μ. From positions 1260–2310 μ there was a dramatic dorsal shift of the PVN cells which by this time had also extended laterally. The total rostral-caudal distance occupied by the PVN cells was 3150 μ. That the lateral-medial distance occupied by the PVN was small (1050 μ) was determined on examining the magnocellular nuclei in sagittal section.
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  • 18
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    Cell & tissue research 240 (1985), S. 315-321 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Fine structure ; Pituitary cell ; Immunocytochemistry ; Goat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The fine structure of each type of anterior pituitary cell in the male goat was studied through the application of a superimposition technique in which adjacent thick sections were used to identify individual cells beforehand by light-microscopic immunohistochemistry. A cone of the pars intermedia protrudes into the pars anterior, being surrounded by the narrow pituitary cleft; the immunohistochemical appearances of the cells forming the cone resemble those of the pars anterior. Several follicles appear in the pars anterior. Ultrastructurally GH cells resemble prolactin cells. The secretory granules of both types are spherical; the diameter of the former is about 340 nm, whereas that of the latter is about 440 nm. ACTH cells are polygonal in shape with secretory granules, about 180 nm in diameter, scattered throughout the cytoplasm. TSH cells, which are spherical in shape, contain the smallest secretory granules, 150 nm in diameter. The highly electron-dense LH cells contain numerous secretory granules about 210 nm in diameter. Their nuclei are irregular with incisures. Thus, the anterior pituitary cells of the goat are ultrastructurally characteristic and species-specific.
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  • 19
    ISSN: 1432-0878
    Keywords: Proctolin ; Immunocytochemistry ; Bioassay ; Leptinotarsa decemlineata ; Peptidergic neuron
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Evidence is presented that neurons in the adult Colorado potato beetle contain a proctolin-like substance. By use of immunocytochemical methods the location of immunoreactive neurons in the central and stomatogastric nervous systems is described. No such neurons were found in the proto- and deutocerebrum or optic lobe. Few immunoreactive neurons are present in the tritocerebrum and numerous proctolin-immunoreactive neurons occur in all ventral ganglia and in the frontal ganglion. Two groups of neurosecretory cells in the suboesophageal ganglion contain a proctolin-immunoreactive substance. In these cells this material is co-localized with a bovine pancreatic polypeptide/FMRF amide-like substance and with a vasopressin/vasotocin/oxytocin-like substance. Proctolin-immunoreactive axon terminals were found on the musculature of the fore- and hindgut and of the vas deferens, and on some segmental muscles. Furthermore, proctolin-immunoreactive neurosecretory axon terminals were found in the corpus cardiacum. The proctolin-like substance may therefore function both as a neurotransmitter/neuromodulator and as a neurohormone. The presence of a proctolin-like substance was also demonstrated with a sensitive bioassay. On fractionation of extracts of the nervous systems of Leptinotarsa decemlineata with high performance liquid chromatography most of the proctolin-like bioactive material comigrated with authentic proctolin. This shows that a proctolin-like substance in this insect is very similar to, if not identical with, the known pentapeptide proctolin.
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  • 20
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    Cell & tissue research 240 (1985), S. 593-600 
    ISSN: 1432-0878
    Keywords: Serotonin ; Immunocytochemistry ; Visceral nerves ; Neurohemal system ; Periplaneta americana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of the indirect immunofluorescent technique applied to whole mounts of tissues and organs of the American cockroach, Periplaneta americana, serotonin-like reactivity has been demonstrated in an extensive meshwork of fibers on the surface of nerves of the subesophageal ganglion and the tritocerebrum. This meshwork appears to serve the neurohemal release of serotonin. In addition, several of these nerves contain two or more centrally located axons that appear to be serotonergic. The corpora cardica show immunoreactivity but do not appear to be a principal release site for serotonin. The nerves and ganglia of the stomadeal nervous system show immunoreactivity for serotonin and provide serotonin-positive innervation to the salivary glands and the visceral muscles of the stomodeum and midgut. The heart, hindgut, Malpighian tubules, fat body, and skeletal muscles all appear to lack serotonin immunoreactivity.
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  • 21
    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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  • 22
    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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  • 23
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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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  • 24
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    Cell & tissue research 251 (1988), S. 433-439 
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone ; Immunocytochemistry ; Pituitary gland ; White- and black-background adaptation ; Teleost, Anguilla anguilla
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Eels were adapted to black- or white-coloured backgrounds and the pituitary glands were prepared for light and electron microscopy. Immunocytochemical staining was used to study the distribution of the neurohypophysial melanin-concentrating hormone in the neurointermediate lobe. The hormone was located in small, elliptical, electron-opaque neurosecretory granules, measuring approximately 120×90 nm. The neurones terminated on blood vessels in the centre of the neurohypophysis and on the basement membrane separating neural and intermediate lobe tissues. The results of both light and electron immunocytochemistry and of radioimmunoassay are consistent with a higher rate of hormone release from eels adapted to white backgrounds than from those adapted to black backgrounds. In addition to this, when fish that had been adapted to white tanks were transferred to black tanks, there was an accumulation of irMCH in the gland and an increased numerical density of secretory granules at nerve terminals. These results reinforce the proposal that MCH is released during adaptation to a white background, to cause melanin concentration and to inhibit MSH release, and that its release is halted in black-adapted fish.
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  • 25
    ISSN: 1432-0878
    Keywords: Pigment-dispersing hormone ; FMRFamide ; Immunocytochemistry ; Carcinus maenas ; Orconectes limosus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of antisera raised against synthetic pigment-dispersing hormone (PDH) of Uca pugilator and FMRFamide, the distribution of immunoreactive structures in the central nervous system (CNS) of Carcinus maenas and Orconectes limosus was studied by light microscopy. In both species, a total of 10–12 PDH-positive perikarya occur amongst the anterior medial, dorsal lateral and angular somata of the cerebral ganglion (CG). In C. maenas, one PDH-perikaryon was found in each commissural ganglion (COG) and several more in the thoracic ganglion. In O. limosus, only four immunopositive perikarya could be demonstrated in the ventral nerve cord, i.e., two somata in the anterior and two in the posterior region of the suboesophageal ganglion (SOG). PDH-immunoreactive tracts and fiber plexuses were present in all central ganglia of both species, and individual axons were observed in the connectives. FMRFamide-immunoreactivity was studied in O. limosus only. Neurons of different morphological types were found throughout the entire CNS, including numerous perikarya in the anterior medial, anterior olfactory, dorsal lateral and posterior cell groups of the CG. Four perikarya were found in the COG, six large and numerous smaller ones in the SOG, and up to eight cells in each of the thoracic and abdominal ganglia. In each ganglion, the perikarya form fiber plexuses. Axons from neurons belonging to the CG could be traced into the ventral nerve cord; nerve fibers arising from perikarya in the SOG appeared to project to the posterior ganglia. In none of the structures examined colocalization of PDH- and FMRF-amide-immunoreactivity was observed.
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  • 26
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    Cell & tissue research 254 (1988), S. 119-124 
    ISSN: 1432-0878
    Keywords: Choline acetyltransferase ; Immunocytochemistry ; Light and electron microscopy ; Supraoptic nucleus ; Paraventricular nucleus ; Rat (Wistar, Long Evans, Brattleboro)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two different monoclonal antibodies raised against choline acetyltransferase were used, together with preembedding immunocytochemical techniques, to visualize the possible cholinergic innervation of the supraoptic and paraventricular nuclei of the rat hypothalamus. Light microscopy confirmed the presence of a group of bipolar and multipolar immunoreactive neurones in the hypothalamus dorsolateral to the supraoptic nucleus as well as numerous immunopositive fibers. Electron microscopy showed that the immunopositive cell bodies contained the usual perikaryal organelles while most immunoreactive fibers appeared dendritic; immunonegative terminals made synaptic contact onto these profiles. Immunopositive terminals making synaptic contact onto dendritic profiles were also noted in this area. In contrast, light microscopy showed no immunoreactivity to choline acetyltransferase in the magnocellular nuclei themselves. Electron microscopy revealed some immunopositive profiles along the boundaries of both nuclei, along the optic chiasm adjacent to the supraoptic nucleus and in the ventral glial lamina but not within the nuclei proper. Surprisingly, these immunopositive profiles appeared dendritic and were often contacted by one or more immunonegative synapses. Our observations thus indicate that cell bodies and dendrites in the supraoptic and paraventricular nuclei are not directly innervated by cholinergic synapses. The functional significance of the putative cholinergic dendrites in close proximity to magnocellular neurones remains to be determined.
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  • 27
    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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  • 28
    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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  • 29
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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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  • 30
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Serotonin ; Diphyllobothrium dendriticum ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present immunocytochemical study concerns the distribution of four neuropeptides, FMRF-amide, vasotocin, leu-enkephalin and neurotensin, and of the bioamine serotonin in the plerocercoid larva of Diphyllobothrium dendriticum. Anti-FMRF-amide and vasotocin-reactivity occurs in perikarya and nerve fibres in the CNS and PNS of this worm. The peptide-containing fibres surround and seem to innervate the musculature and to terminate beneath the basal lamina of the tegument at the inner surface of the bothridia, suggesting a neurotransmitter function. Antileu-enkephalin reaction occurs in perikarya and fibres in the main nerve cords and in the PNS. Anti-neurotensin reactive fibres were observed in the neuropile of the nerve cords. Serotonin immunoreactivity was found in neurons in the ganglionic commissure of the brain and along the main nerve cords. This study is the first immunocytochemical identification of neuropeptides and serotonin in a parasitic flatworm and the information gained may be of importance for the development of new antihelminthics.
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  • 31
    ISSN: 1432-0878
    Keywords: Nervous system ; Insects ; Immunocytochemistry ; FMRFamide ; Bovine pancreatic polypeptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of FMRFamide immunoreactive neurones in the ventral nerve cord of the locust, Schistocerca gregaria, is described. These neurones are found only in the suboesophagael and thoracic ganglia, although immunoreactive processes are found in the neuropils of the abdominal ganglia. Many of these neurones also react with an antiserum raised against bovine pancreatic polypeptide (BPP), but this antiserum also reveals another population of cells in the abdominal ganglia. The staining obtained with the BPP antiserum is blocked by preabsorption of the antiserum with FMRFamide; the converse is not true: FMRFamide-like immunoreactivity is not suppressed by preincubation with BPP. These results suggest that there are at least two endogenous peptide antigens in the locust nerve cord: one is found in cells of the suboesophageal and thoracic ganglia, and the other is found in cells of the abdominal ganglia.
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  • 32
    ISSN: 1432-0878
    Keywords: Somatostatin ; Immunocytochemistry ; Brain ; Pituitary ; Teleost fish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An extensive system of somatostatin-immunoreactive neurons has been localized in the forebrain and pituitary of the molly (Poecilia latipinna), using the unlabelled antibody immunocytochemical method. In the hypothalamus, reactive perikarya were scattered throughout the parvocellular divisions of the preoptic nucleus. These cells were smaller in size and more ventral in position than those which stained with antisera to the neurohypophysial hormones, vasotocin and isotocin. A few very small somatostatin-immunoreactive cells were observed in the tuberal region and in the nuclei of the lateral and posterior recesses — areas which were rich in somatostatin-immunoreactive fibres. Somatostatin cells were also found in a small area of the ventral thalamus, mainly in the dorsolateral nucleus. Some of these neurons were large and multipolar, and appeared to form tracts of fibres into the posterior hypothalamus. In the telencephalon there were a few stained cells in the ventral area, with a complex pattern of fibres occurring in parts of the dorsal area. Somatostatin-immunoreactivity was intense in the central and posterior neurohypophysis, and particularly in its finger-like projections into the proximal pars distalis, around groups of growth hormone cells. Examination of material from fishes under various experimental conditions provided evidence for the somatostatin fibres originating from the preoptic neurons being involved in the control of growth hormone secretion.
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  • 33
    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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  • 34
    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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  • 35
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    Cell & tissue research 254 (1988), S. 517-530 
    ISSN: 1432-0878
    Keywords: Uterus ; Autonomic innervation ; Neuropeptides ; Immunocytochemistry ; Retrograde tracing ; Pregnancy ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The occurrence and distribution of peptidergic nerves in the guinea pig uterus was studied by means of immunocytochemistry using numerous neuropeptide antisera. Neuropeptide Y (NPY)-immunoreactive (IR) nerves were the most abundant, whereas substance P (SP)-, calcitonine gene-related peptide (CGRP)-, and neurokinin A (NKA)-IR nerves were less frequent, and peptide histidine isoleucine (PHI)-IR nerves were the most sparse. Chemical sympathectomy by means of 6-hydroxydopamine, and capsaicin treatment revealed the division of the peptidergic nerves into three separate populations: (1) NPY-IR nerves, which co-existed with adrenergic nerves, (2) SP-, CGRP-and NKA-IR nerves, which mutually co-existed, and (3) PHI-IR nerves. Parallel-running adrenergic/NPY-IR and SP-IR nerves could be found with very similar although not completely identical morphological appearance. Paracervical ganglia contained neurotensin-and dynorphin A-IR cell bodies in addition to cell bodies with immunoreactivities similar to those in prevertebral ganglia. Combined retrograde tracing with True blue and immunocytochemistry showed that the adrenergic and NPY-IR uterine nerves originate in paracervical and prevertebral ganglia. In the prevertebral ganglia the cellular origin was the same for adrenergic and NPY-IR nerves. In contrast, SP-, CGRP-,and NKA-IR nerves originated in dorsal root ganglia. At full-term pregnancy all the neuropeptide immunoreactivities had vanished, probably reflecting a fetus-induced general nerve degeneration.
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  • 36
    ISSN: 1432-0878
    Keywords: Pineal complex ; Pinealocytes, receptor line ; Subcommissural organ ; Immunocytochemistry ; Protein secretion ; Neuroendocrine system Geotria australis (Cyclostomata) ; Onkorhynchus kisutch (Teleostei) ; Eupsophus roseus (Anura) ; Heloderma suspectum, Varanus monitor (Lacertilia) ; Domestic fowl ; Rat ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of light-microscopic immunocyto-chemistry two polyclonal antibodies (AFRU, ASO; see p. 470) directed against secretory glycoproteins of the subcom-missural organ were shown to cross-react with cells in the pineal organ of lamprey larvae, coho salmon, a toad, two species of lizards, domestic fowl, albino rat and bovine (taxonomic details, see below). The AFRU-immunoreactive cells were identified as pinealocytes of the receptor line (pineal photoreceptors, modified photoreceptors or classical pinealocytes, respectively) either due to their characteristic structural features or by combining AFRU-immunoreaction with S-antigen and opsin immunocytochemistry in the same or adjacent sections. Depending on the species, AFRU- or ASO-immunoreactions were found in the entire perikaryon, inner segments, perinuclear area, and in basal processes facing capillaries or the basal lamina. In most cases, only certain populations of pinealocytes were immunolabeled; these cells were arranged in a peculiar topographical pattern. In lamprey larvae, immunoreactive pinealocytes were observed only in the pineal organ, but not in the parapineal organ. In coho salmon, the immunoreaction occurred in S-antigen-positive pinealocytes of the pineal end-vesicle, but was absent from S-antigen-immunoreactive pinealocytes of the stalk region. In the rat, AFRU-immunoreaction was restricted to S-antigen-immunoreactive pinealocytes found in the deep portion of the pineal organ and the habenular region. These findings support the concept that several types of pinealocytes exist, which differ in their molecular, biochemical and functional features. They also indicate the possibility that the AFRU- and ASO-immunoreactive material found in certain pinealocytes might represent a proteinaceous or peptidic compound, which is synthesized and released from a specialized type of pinealocyte in a hormone-like fashion. This cell type may share functional characteristics with peptidergic neurons or paraneurons.
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  • 37
    ISSN: 1432-0878
    Keywords: Enkephalin-related peptides ; Immunocytochemistry ; Neuropeptides ; Co-existence of peptides ; Neurosecretory cells ; Blowly, Calliphora vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of enkephalin-like immunoreactive material has been studied in the CNS of C. vomitoria. The presence of both Met- and Leu-enkephalin-related peptides is suggested by differential immunostaining with a variety of antisera. Comparisons made between certain of the enkephalin-immunoreactive perikarya, nerve fibres and terminals with cells in corresponding positions as evidenced in previously published neuroanatomical studies of the dipteran brain have suggested specific enkephalinergic pathways. As examples, one Met-enkephalin-immunoreactive neuron appears to link the lobula with the dorsal protocerebrum, and a group of Leu-enkephalin cells in the pars intercerebralis appear to have arborisations in both the central body (fan-shaped body) and the tritocerebral neuropil around the oesophageal foramen. Neuronal pathways of this type indicate that the enkephalin-like peptides of the fly brain are functioning as neurotransmitters and/or neuromodulators. In the thoracic ganglia, symmetrically arranged cells, immunoreactive to both Met- and Leu-enkephalin antisera, are positioned ventrally in pairs on either side of the mid-line in a sagittal plane. Very little immunoreactive material is observed in the neuropil, however, and the source of the accumulation of Leu-enkephalin-immunoreactivity in the dorsal neural sheath is not certain. It is suggested that this material, in contrast to that present in areas of the brain, acts as a neurohormone and that it may have a physiological role following its release into the haemolymph. The enkephalin-like immunoreactive material of certain neurons identified within the brain and thoracic ganglion shows a complex pattern of co-existence with pancreatic polypeptide- and gastrin/cholecystokinin-like peptides.
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  • 38
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    Cell & tissue research 251 (1988), S. 441-449 
    ISSN: 1432-0878
    Keywords: Thyrotropin-releasing hormone (TRH) ; Immunocytochemistry ; Neuropeptides ; Avian brain ; Domestic mallard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of immunoreactive thyrotropin-releasing hormone (TRH) in the central nervous system of the domestic mallard was studied by means of the peroxidase-antiperoxidase technique. After colchicine pretreatment, the highest number of TRH-immunoreactive perikarya was found in the parvocellular subdivision of the paraventricular nucleus and in the preoptic region; a smaller number of immunostained perikarya was observed in the lateral hypothalamic area and in the posterior medial hypothalamic nucleus. TRH-immunoreactive nerve fibers were detected throughout the hypothalamus, forming a dense network in the periventricular area, paraventricular nucleus, preoptic-suprachiasmatic region, and baso-lateral hypothalamic area. TRH-containing nerve fibers and terminals occurred in the organon vasculosum of the lamina terminalis and in the external zone of the median eminence in juxtaposition with hypophyseal portal vessels. Scattered fibers were also seen in the internal zone of the median eminence and in the rostral portion of the neural lobe. Numerous TRH-immunoreactive fibers were detected in extra-hypothalamic brain regions: the highest number of immunoreactive nerve fibers was found in the lateral septum, nucleus accumbens, olfactory tubercle, and parolfactory lobe. Moderate numbers of fibers were located in the basal forebrain, dorsomedial thalamic nuclei, hippocampus, interpeduncular nucleus, and the central gray of the mesencephalon. The present findings suggest that TRH may be involved in hypophysiotropic regulatory mechanisms and, in addition, may also act as neuromodulator or neurotransmitter in other regions of the avian brain.
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  • 39
    ISSN: 1432-0878
    Keywords: Posterior pituitary ; Immunocytochemistry ; Anti-GABA ; GABA-transaminase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antibody against gamma-aminobutyric acid (GABA) was used to identify GABAergic elements immunocytochemically in the rat posterior pituitary. In order to increase the intracellular concentration of GABA, rats were treated with the GABA-transaminase inhibitor gamma-vinyl-GABA (GVG). Light-microscopic observations of Vibratome and semithin sections revealed the presence of numerous immunoreactive nerve fibers throughout the neural lobe; the mean number and length of these fibers increased by 90% after GVG treatment. Electron microscopy demonstrated the immunostained axons to be of small diameter. The reaction product was confined to small vesicles. No immunostaining occurred in pituicytes. The richness of the GABAergic innervation of the neural lobe contrasts with previous reports using antibodies against glutamate decarboxylase and supports the idea that GABA participates in the presynaptic control of neurosecretion.
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  • 40
    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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  • 41
    ISSN: 1432-0878
    Keywords: Substance P-related peptides ; Immunocytochemistry ; Invertebrate ganglia ; Peptidergic neurons ; Stomatogastric nervous system ; Cancer borealis, Panulirus interruptus, Homarus americanus (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of substance P-like immunoreactivity in the stomatogastric nervous systems of three decapod crustacean species, Cancer borealis, Homarus americanus, and Panulirus interruptus, was studied. The stomatogastric ganglion showed dense staining in the neuropil, but none in the somata. A single neuron stained in the esophageal ganglion. Lucifer yellow backfills and intracellular injections followed by incubation with the substance P antibody showed that the axons of this neuron project into the inferior esophageal nerves towards the paired commissural ganglia. The commissural ganglia showed a pronounced projection from a large bundle of fibers in the anterior medial portion of the circumesophageal connective. Additionally, less dense neuropil and stained somata were seen in the commissural ganglia. Staining was completely blocked by preabsorption with authentic substance P, physalaemin, eledoisin, and substance K. These data suggest that in the nervous system of crustacean species a molecule with C-terminal homology to substance P and other tachykinins is released as a neuroregulator in the stomatogastric ganglion.
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  • 42
    ISSN: 1432-0878
    Keywords: Prolactin-like neuropeptides ; Immunocytochemistry ; Brain ; Neuroendocrine structures ; Insects ; Leucophaea maderae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of prolactin-like neuropeptides was demonstrated immunocytochemically in the brain and affiliated neuroendocrine structures of the insect Leucophaea maderae. Use of the unlabelled peroxidase-antiperoxidase method of Sternberger revealed a rather widespread and differential distribution of reaction products resembling human (hPRL) and ovine (oPRL) prolactin. Tests with antirat PRL antibody were negative. The specificity of the antibodies used was established by liquid-phase absorptions and confirmed in tissue control systems. In L. maderae, anti-oPRL identifies part of an oPRL-like molecule different from human and rat PRL. Anti-hPRL reveals part of a human and ovine PRL-like molecule different from rat prolactin. These results indicate the occurrence, in the nervous tissue of one insect species, of at least two types of prolactin-like molecules.
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  • 43
    ISSN: 1432-0878
    Keywords: Collagen ; Carboxypropeptide ; Chondrocyte ; Tissue culture ; Immunocytochemistry ; Immunofluorescence ; Chick embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antibody reacting with the C-propeptide of chick type-II procollagen was used in an attempt to localize this terminal extension of the procollagen molecule (by immunogold labelling) during early collagen fibrillogenesis in chondrocyte cultures. After 2 days in culture the chondrocytes were surrounded by pericellular type-II collagen, as demonstrated by an indirect immunofluorescence labelling technique. An electron microscopy study of these cultures showed that the collagen fibrils were thin (∼ 15 nm diameter), with a poorly visible cross striation, sometimes enhanced by slight thickenings. The antibody against the C-propeptide of type-II procollagen labelled most of the collagen fibrils, according to a very regular pattern constituting a 60 nm periodicity. After 3 days the label was still present on the pericellular collagen fibrils but disappeared from the collagen fibrils of the extracellular matrix. Our results indicate that the C-propeptide of type-II procollagen is retained in the newly formed fibrils.
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  • 44
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    Cell & tissue research 188 (1978), S. 259-264 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Human fetus ; Oxytocin ; Neurophysin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The use of antibodies against oxytocin or neurophysin enabled the detection by immunocytochemistry of oxytocin-neurophysin neurons in the hypothalamus in the human fetus. The perikarya of these neurons are located in the paraventricular and supraoptic nuclei. Immunoreactive neurons occur in the median eminence. The neurophysin immunoreactive neurons were more numerous than the oxytocin immunoreactive neurons. The specificity of the immunocytological reaction was controlled. The first oxytocin-neurophysin neurons are seen as early as the 14th week of gestation.
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  • 45
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Enkephalins ; Neurohemal organ ; Carcinus maenas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light-microscopical observations with immunofluorescence and peroxidase staining procedures revealed leu-enkephalin-like immunoreactivity in axon profiles of the sinus gland (SG) and in single small neurons in the optic ganglia of the eyestalk of Carcinus maenas. Electron microscopy of the SG showed reactivity to be associated with neurosecretory granules 82±23 nm in diameter. High performance liquid chromatography of SG-extracts revealed radioimmunoreactive substances with the retention times of synthetic met- and leu-enkephalin and met-enkephalin-Arg6-Phe7, respectively.
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  • 46
    ISSN: 1432-0878
    Keywords: S-100 protein ; Folliculo-stellate cells ; Normal human pituitary, anterior lobe ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With the use of an anti-human S-100 protein antibody, it was possible to reveal a characteristic cell type in the anterior lobe of the normal human pituitary. These cells, so-called folliculo-stellate cells, were present in all pituitaries studied but their number varied from one gland to another. Immunoreactive cells, isolated or grouped, were arranged close to various secretory granulated cells. Especially by use of double immunoenzymatic labeling, it was evident that these cells are spatially related either to somatotropes, prolactin cells and “corticotropes”, or to glycoprotein-containing cells. Such immunoreactive cells were rare or absent in pseudo-follicular arrangements of secretory granulated cells. Since it is now possible to identify this cell type by light microscopy and since no reliable functional significance is known, it seems more advisable to term this cell type “stellate cell” instead of “folliculostellate cell”.
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  • 47
    ISSN: 1432-0878
    Keywords: Reissner's fiber ; Subcommissural organ ; Ependyma ; Immunocytochemistry ; Electron microscopy ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Reissner's fiber (RF) of the subcommissural organ (SCO), the central canal and its bordering structures, and the filum terminale were investigated in the bovine spinal cord by use of transmission electron microscopy, histochemical methods and light-microscopic immunocytochemistry. The primary antisera were raised against the bovine RF, or the SCO proper. Comparative immunocytochemical studies were also performed on the lumbo-sacral region of the rat, rabbit, dog and pig. At all levels of the bovine spinal cord, RF was strongly immunoreactive with both antisera. From cervical to upper sacral levels of the bovine spinal cord there was an increasing number of ependymal cells immunostainable with both antisera. The free surface of the central canal was covered by a layer of immunoreactive material. At sacral levels small subependymal immunoreactive cells were observed. From all these structures sharing the same immunoreactivity, only RF was stained by the paraldehyde-fuchsin and periodicacid-Schiff methods. At the ultrastructural level, ependymal cells with numerous protrusions extending into the central canal were seen in the lower lumbar segments, whereas cells displaying signs of secretory activity were principally found in the ependyma of the upper sacral levels. A few cerebrospinal fluid-contacting neurons were observed at all levels of the spinal cord; they were immunostained with an anti-tubulin serum. The lumbo-sacral segments of the dog, rat and rabbit, either fixed by vascular perfusion or in the same manner as the bovine material, did not show any immunoreactive structure other than RF. The possibilities that the immunoreactive ependymal cells might play a secretory or an absorptive role, or be the result of post-mortem events, are discussed.
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  • 48
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    Cell & tissue research 240 (1985), S. 735-741 
    ISSN: 1432-0878
    Keywords: Retina, chicken ; Photoreceptor outer segments ; Photopigments ; Immunological heterogeneity ; Antibodies ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immune cells producing antibodies to chicken photoreceptor membranes were fused with myeloma cells and supernatants of the resulting hybridoma cells were used to test various types of photoreceptor cells in the chicken retina by means of immunocytochemistry. A polyclonal antibody raised against the protein component of bovine rhodopsin was also used. Outer segments of various photoreceptor cells were labelled by the following antibodies: rods were positive with the anti-rhodopsin antibody, both members of the double cones were stained by supernatant A1, while one type of single cones (designated as type A) was specifically labelled by supernatants A5, B3 and D6. The other type of single cones (type B) reacted with anti-rhodopsin and supernatant A1. The results indicate that there are distinct differences in the molecular structure of various photoreceptor outer segments.
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  • 49
    ISSN: 1432-0878
    Keywords: Indoleamines ; Serotonin ; Retina (turtle) ; Amacrine/bipolar cells ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using immunocytochemical methods, we have been able to demonstrate serotonin-like immunoreactivity (SLI) in amacrine and bipolar cells of the turtle retina. Inhibition of monoamine oxidase with pargyline drastically increases the amount of 5-hydroxytryptamine within both cell types. The indoleamine 6-hydroxytryptamine is taken up by both cell types and both types are destroyed within 10 days following intraocular injection of 5,7-dihydroxyryptamine. Increasing the external potassium concentration induces release of serotonin in both cell types. Our data support the idea that these neurons use serotonin during neuronal processing. Morphologically, amacrine and bipolar cells with SLI can be subdivided into two and three subclasses, respectively, based on their ramification pattern within the inner plexiform layer. A comparison of the morphological data with those of intracellularly stained amacrine and bipolar cells suggests that all bipolar cells with SLI are center-hyperpolarizing cells and all amacrine cells center-depolarizing cells.
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  • 50
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    Cell & tissue research 241 (1985), S. 391-397 
    ISSN: 1432-0878
    Keywords: Fibronectin ; Gastrulation ; Immunocytochemistry ; Ultrastructure ; Basal lamina ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural distribution of fibronectin immunoreactivity was investigated in the chicken embryo during late gastrulation. Sites of binding of anti-fibronectin antibodies were ascribed to the basal lamina and associated structures, and to the cell surface. The fibronectin-rich basal lamina was resolved into (1) a lamina densa, which appears as a continuous, dense sheet, (2) a lamina lucida, consisting of anchoring cords between lamina densa and epithelial cells, and (3) a lamina intima, closely juxtaposed to the cell surface. Cell-surface labelling was also observed in mesoblast cells, and along the dorsal side of the deep-layer cells. The ventral side of the latter cells was poorly stained in the endophyllic crescent, except in coated pits, and more regularly stained at the level of definitive endoblast. Some structures associated with the basal lamina reacted intensely with anti-fibronectin antibodies. These are (1) the interstitial bodies, which are aggregates of extracellular material, and (2) a kind of fibril or tubule, embedded in a fibronectin matrix and mainly found in the endophyllic crescent. Some intracellular labelling was found in most deep-layer cells, in few epiblast cells, never in mesoblast cells. These results extend previous studies on the localization of fibronectin, and correlate its presence and surface topology with its postulated role in migration of mesoblast cells on the basal lamina which, chemically, constitutes an appropriate substrate.
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  • 51
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Hibernation ; Pars distalis ; Miniopterus schreibersii fuliginosus (Chiroptera)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical studies were performed to describe the characteristics of cell types and their distribution in the pars distalis of Japanese long-fingered bat, Miniopterus schreibersii fuliginosus, collected at various stages of the reproductive cycle. Six distinct cell types have been identified in the pars distalis by the unlabeled immunoperoxidase technique and by the ABC method. Growth hormone (GH) and prolactin (PRL) cells were immunostained with antisera against chicken GH and ovine PRL. The GH-immunoreactive cells were round or oval orangeophilic cells distributed throughout the pars distalis with prominent aggregation in the posterolateral region. The PRL cells were pleomorphic carminophilic cells that occurred in small groups within the central and dorsocaudal regions of the pars distalis. They were sparsely distributed in the central region of the pars distalis in the hibernating bats, but increased significantly in the pregnant and lactating bats. The adrenocorticotropic (ACTH) cells were large round or polygonal amphophilic cells in the rostroventral and ventrolateral regions of the pars distalis. The thyrotropic (TSH) cells were small rounded or polygonal and distributed mainly in the ventrolateral region of the pars distalis. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) cells were identified immunocytochemically with antisera against the specific beta subunits of ovine LH and rat FSH. There were two populations of LH and FSH cells, one aggregated in the zona tuberalis and the other scattered singly throughout the rest of the pars distalis. The aggregated cells were immunoreactive with both antisera directed to LH and FSH, while scattered cells were reactive solely with antiserum to either LHβ or FSH and exhibited seasonal variations. In females, the proportional volume of the pars distalis occupied by LH cells was significantly reduced during pregnancy and lactation. No evidence of involution was observed in pars distalis cells except for PRL cells in males or females during hibernation.
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  • 52
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    Cell & tissue research 251 (1988), S. 307-313 
    ISSN: 1432-0878
    Keywords: Lysozyme (muramidase) ; Paneth cell ; Gastric gland ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of lysozyme in normal gastric and duodenal mucosa was studied by light- and electronmicroscopic immunocytochemical techniques (direct enzyme-labeled antibody method). In the duodenal mucosa, lysozyme was found in the Paneth cells and the epithelial cells of Brunner's glands. Electron-microscopically, lysozyme was found in rough endoplasmic reticulum and perinuclear spaces, which were assumed to be protein-synthesizing organelles, and also in the secretory granules of Paneth cells. Additionally, lysozyme was detected in the stomach in mucinous granules and in some parts of the rough endoplasmic reticulum within the epithelial cells of the pyloric glands, the mucous neck cells of the fundic glands, and in several surface epithelial cells of the plyoric and fundic regions. This suggests that some quantity of lysozyme in gastrointestinal secretion originates from the gastric and duodenal glands, and that it acts as a defense mechanism in the gastrointestinal tract.
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  • 53
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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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  • 54
    ISSN: 1432-0878
    Keywords: Melanin-concentrating hormone (MCH) ; Hypothalamus ; Melanin-concentrating activitiy ; Radioimmunoassay ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Melanin-concentrating hormone (MCH) is a neurosecretory peptide that induces melanin concentration within teleost melanophores. Here, we characterized MCH-like substance in the rat brain by both an in vitro fish-scale melanophore bioassay and a radioimmunoassay with a salmon MCH antiserum that is directed toward the carboxy-terminus and requires the cyclic configuration for recognition. Furthermore, subcellular localization of the MCH in the rat brain was examined by immunocytochemistry using electron microscopy. We confirmed that MCH-immunoreactivity and MCH-bioactivity were present together in the same effluent fractions of the rat hypothalamic extracts by reverse-phase high-performance liquid chromatography (HPLC). At electron microscopic level, MCH-immunoreactivity was located specifically in secretory granules in MCH-positive cell bodies confined to the hypothalamus with their neuronal processes projecting widely in the rat brain. Although full characterization of substance must await its isolation, our results strongly support the notion that rat MCH-like substance may be homologous but not identical to salmon MCH, and simultaneously may serve some neurotransmitter and/or neuromodulator role in the brain of the rat.
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  • 55
    ISSN: 1432-0878
    Keywords: Moult-inhibiting hormone ; Hyperglycemic hormone ; Immunocytochemistry ; Neurosecretion ; Decapod crustaceans (five species)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of antisera raised against purified moultinhibiting (MIH) and crustacean hyperglycemic hormone (CHH) from Carcinus maenas, complete and distinct neurosecretory pathways for both hormones were demonstrated with the PAP and immunofluorescence technique. By double staining, employing a combination of silver-enhanced immunogold labelling and PAP, both antigens could be visualized in the same section. Immunoreactive structures were studied in Carcinus maenas, Liocarcinus puber, Cancer pagurus, Uca pugilator and Maja squinado. They were only observed in the X-organ sinus gland (SG) system of the eyestalks and consisted of MIH-positive perikarya, which were dispersed among the more numerous CHH-positive perikarya of the medulla terminalis X-organ (XO). The MIH-positive neurons form branching collateral plexuses adjacent to the XO and axons that are arranged around the CHH-positive central axon bundle of the principal XO-SG tract. In the SG, MIH-positive axon profiles and terminals, clustered around hemolymph lacunae, are distributed between the more abundant CHH-positive axon profiles and terminals. Colocalisation of MIH and CHH was never observed. The gross morphology of both neurosecretory systems was similar in all species examined, however, in U. pugilator and M. squinado immunostaining for MIH was relatively faint unless higher concentrations of antiserum were used. Possible reasons for this phenomenon as well as observed moult cycle-related differences in immunostaining are discussed.
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  • 56
    ISSN: 1432-0878
    Keywords: Corticotropin-releasing hormone ; Somatostatin ; Oxytocin ; Vasopressin ; Luteinizing hormone-releasing hormone ; Immunocytochemistry ; Reproductive cycle ; Ferret ; Mink
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothalamic systems secreting corticotropin-releasing hormone (CRF), somatostatin, oxytocin, vasopressin and luteinizing hormone-releasing hormone (LHRH) were characterized using immunochemistry, and variations were studied in relation to the recrudescence of testicular activity in the ferret and the mink, two species with opposite photoregulation of their annual reproductive cycles. Under the present conditions of study, the immunoreactivity of the CRF, somatostatin, and oxytocin systems showed no significant variation in either species. In contrast, in these two species, the immunoreactivity of the LHRH system varied considerably depending on the date of observation. The increase in the number and immunoreactivity of the LHRH-secreting neurons that occurred in November in the mink and in January in the ferret, is in agreement with previous results showing that the photoperiod plays an essential role in regulating the annual activity of the testis and that the photoperiodic environmental conditions required for the activation of the LHRH system differ between the species. Similarly, correlations could be found between an increase in immunoreactivity of the vasopressinergic axons projecting to the external median eminence and the recrudescence of testicular activity.
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  • 57
    ISSN: 1432-0878
    Keywords: Lipoprotein ; Muscle ; Extracellular space ; Cryosectioning ; Immunocytochemistry ; Locust (Locusta migratoria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Locust lipoproteins (lipophorins) were localized by indirect immunofluorescence- and immunogold labelling in cryosections of dorsolongitudinal flight muscles. Immunolabelling was performed with monoclonal antibodies against apolipoprotein epitopes that are exposed at the surfaces of the lipophorin particles. Both at rest and during flight, lipophorins were located only in the wider spaces of the extracellular matrix, in the basement membranes of the individual muscle fibers and in the extracellular spaces that surround interfibrillar tracheoles. No internalization of lipophorins by the flight muscle cells was observed. Our results indicate that the unloading of lipophorins at the flight muscles is an extracellular event. Similarities with the vertebrate system of chylomicron and very-low-density lipoprotein degradation are discussed.
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  • 58
    ISSN: 1432-0878
    Keywords: Substance P ; Enkephalin ; Spinal dorsal horn ; Immunocytochemistry ; Electron microscopy ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A preembedding dual immunolabeling technique and electron microscopy were utilized to demonstrate the localization of immunoreactive substance P and methionine-enkephalin-octapeptide (Enk-8) in ultrathin sections of the surface layer (laminae I and II) of rat spinal dorsal horn. The immunoreaction of Enk-8 was visualized as goldtoned silver particles and that of substance P as diaminobenzidine reaction products. Axonal terminals with immunoreactive substance P, and also unlabeled axonal terminals, formed synaptic junctions with the perikarya and dendritic processes of Enk-8-containing neurons. Dendritic profiles immunolabeled for substance P were synaptically linked with unlabeled axons but not with Enk-8-positive ones. Furthermore, it was found that Enk-8 axons and substance P axons terminated synaptically in juxtaposition to one another on the same immunonegative dendrites. Among the Enk-8-containing neurons axonal profiles also appeared to be synaptically associated with immunoreactive Enk-8 dendritic processes.
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  • 59
    ISSN: 1432-0878
    Keywords: Calcitonin ; Parathyroid hormone ; Estradiol ; Estradiol receptors ; Osteoblasts ; Immunocytochemistry ; Cryo-ultramicrotomy ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactivities to endogenous calcitonin, endogenous parathyroid hormone, endogenous estradiol and estradiol receptors were studied in osteoblasts from the calvaria of neonatal mice by immunocytochemistry with the use of ultrathin sections obtained by cryo-ultramicrotomy. Tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and frozen in liquid nitrogen. Estradiol and estradiol receptors could not be detected in osteoblasts, whereas calcitonin- and parathyroid hormone-like immunoreactivities were observed in this cell type. Calcitonin and parathyroid hormone had similar subcellular localizations: immunoreactivities were observed at the plasma-membrane level, in the cytoplasmic matrix, and in the nucleus. These results provide immunocytological evidence for: 1) the internalization of calcitonin and parathyroid hormone in osteoblasts; 2) a direct participation of calcitonin and parathyroid hormone in the regulation of osteoblasts; 3) the absence of estradiol receptors and estradiol in osteoblasts.
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  • 60
    ISSN: 1432-0878
    Keywords: Crustacean hyperglycemic hormone ; Astacus leptodactylus ; Immunocytochemistry ; Day/night rhythmicity ; Neurosecretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The secretory dynamics of the Crustacean Hyperglycemic Hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus were studied during the daily cycle (12 h light/12 h dark). The different secretory stages of individual cells were determined by means of immunocytochemistry combined with morphometric analysis at the light-microscopic level. The data obtained were correlated with the 24-h rhythmicity of blood glucose concentration. The results suggest the following hypothesis. The synthetic activity of the CHH cells receives a stimulus 2 h before the beginning of the dark period, resulting in a pronounced transfer of CHH granules into the axons. These CHH granules reach the axon terminals after the onset of the dark period. At that time a burst of exocytotic activity occurs, causing a strong release of CHH into the hemolymph. Four hours later this CHH release results in hyperglycemia. The same process, though with less intensity, is repeated and causes a second smaller glucose peak at the beginning of the light period.
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  • 61
    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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  • 62
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Enkephalins ; FMRFamide ; Substance P ; Uca pugilator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The occurrence and distribution of substance P (SP)-like, methionine-(Met)- and leucine-(Leu)-enkephalinlike, and FMRFamide-like immunoreactivities were determined in the neuroendocrine complex of the eyestalk of the fiddler crab, Uca pugilator, by immunocytochemistry. SP-like immunoreactivity was found in the optic peduncle, sinus gland, medulla externa, medulla interna, lamina ganglionaris, and retinular cells. Met-enkephalin-like and Leuenkephalin-like immunoreactivity was observed in most of the retinular cells, optic peduncle, sinus gland, medulla terminalis, and lamina ganglionaris. However, Met-enkephalin-like, but no Leu-enkephalin-like, immunoreactivity was seen in the medulla terminalis X-organ. FMRFamide-like immunoreactivity could be seen in all parts of the eyestalk except in the sinus gland, lamina ganglionaris, and retinular cells. FMRF-amide-like activity was especially strong in the three chiasmatic regions connecting the optic ganglia. The possibility that these four peptides may function as neuroregulators in the fiddler crab is discussed.
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  • 63
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Teleost fish ; Pituitary ; Growth hormone releasing hormone ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using an antiserum directed against the C-terminus of hGRH(1–44)NH2 and another recognizing the mid portion to C-terminal of hGRH(1–40)OH, we identify two immunocytochemically distinct GRH-immunoreactive systems in the brain of the codfish, Gadus morhua. The antiserum directed against GRF(1–44)NH2 stains cell bodies exclusively in the rostral pars distalis. The other antiserum immunoreactive with GRF(1–40)OH reacts with a population of parvocellular and magnocellular neuronal cell bodies in the hypothalamus and with two major axonal pathways which project toward the median eminence and terminate primarily in the pars nervosa. These results indicate the presence of at least two forms of hGRH-like peptides in the teleost which may have different roles in the regulation of pituitary function.
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  • 64
    ISSN: 1432-0878
    Keywords: Bioactive peptides ; Coexistence of peptides ; Immunocytochemistry ; Electron microscopy ; Insect nervous system ; Calliphora erythrocephala, C. vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ventral thoracic neurosecretory cells (VTNCs) of the blowflies, Calliphora erythrocephala and C. vomitoria, innervating thoracic neuropil and the dorsal neural sheath of the thoracico-abdominal ganglion have been shown to be immunoreactive to a variety of mammalian peptide antisera. In the neural sheath the VTNC terminals form an extensive neurohaemal network that is especially dense over the abdominal ganglia. The same areas are invaded by separate, ut overlapping serotonin-immunoreactive (5-HT-IR) projections derived from neuronal cell bodies in the suboesophageal ganglion. Immunocytochemical studies with different antisera, applied to adjacent sections at the lightmicroscopic level, combined with extensive cross-absorption tests, suggest that the perikarya of the VTNCs contain co-localized peptides related to gastrin/cholecystokinin (CCK), bovine pancreatic polypeptide (PP), Met- and Leuenkephalin and Met-enk-Arg6-Phe7 (Met-enk-RF). Electron-microscopic immunogold-labeling shows that some of the terminals in the dorsal sheath react with several of the individual peptide antisera, whilst others with similar cytology are non-immunoreactive. In the same region, separate terminals with different cytological characteristics contain 5-HT-IR. Both 5-HT-IR and peptidergic terminals are localized outside the cellular perineurium beneath the acellular permeable sheath adjacent to the haemocoel. Hence, we propose that various bioactive substances may be released from thoracic neurosecretory neurons into the circulating haemolymph to act on peripheral targets. The same neurons may also interact by synaptic or modulatory action in the CNS in different neuropil regions of the thoracic ganglion.
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  • 65
    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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  • 66
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Immunocytochemistry ; Neuropeptides ; Hibernating bat ; Miniopterus schreibersii (Chiroptera)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To elucidate the role of hypothalamic neuropeptides in regulation of reproductive phenomena of seasonally breeding feral mammals, we used Japanese long-fingered bats, Miniopterus schreibersii fuliginosus, for immunocytochemical study of distribution of the following neuropeptides in the hypothalamus: arginin vasopressin, oxytocin, luteinizing hormone-releasing hormone, somatostatin, corticotropin-releasing factor, and growth hormone-releasing factor. The size, shape and location of supraoptic, paraventricular, suprachiasmatic, and arcuate nuclei of the bat were determined. Arginin vasopressin-and oxytocin-immunoreactive magnocellular neurons were found in the supraoptic and paraventricular nuclei, where they exhibited separate distribution into two distinct groups. Parvocellular arginin vasopressin neurons occurred only in the suprachiasmatic nucleus. The hibernating bats exhibited slightly increased numbers of vasopressin and oxytocin neurons in the supraoptic and paraventricular nuclei. The pregnant bat displayed further increased numbers of vasopressin and oxytocin neurons in both nuclei. Somatostatin-immunoreactive neurons in the paraventricular nucleus were also immunopositive to anti-oxytocin serum, while those in the ventromedial and arcuate nuclei reacted solely to anti-somatostatin serum. They projected to the anterior median eminence and infundibular stalk. Luteinizing hormone-releasing hormone-immunoreactive perikarya were scattered throughout the basal hypothalamus, being particularly abundant in the arcuate nucleus. They were larger in size in hibernating bats than those in normal (non-pregnant) and pregnant females. They projected fibers mainly to the internal layer of the median eminence and infundibular stalk. A few luteinizing hormone-releasing hormone-reactive fibers were also observed in the organum vasculosum laminae terminalis, lateral habenular nuclei, pineal stalk, retroflexus fasciculus, and olfactory tubercle. Corticotropin releasing factor-immunoreactive perikarya were distributed in the paraventricular nucleus and medial preoptic area and projected into the external layer of the anterior median eminence, while growth hormone-releasing factor-immunoreactive perikarya occurred only in the arcuate nucleus and projected into the posterior part of the median eminence.
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  • 67
    ISSN: 1432-0878
    Keywords: Corpus cardiacum ; Serotonin ; Immunocytochemistry ; Lucifer yellow ; Double labelling ; Locusta migratoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The serotoninergic innervation of the corpus cardiacum (CC) of Locusta migratoria was investigated using two antisera against serotonin. A dense network of immunoreactive nerve fibres was present in the storage lobe of the CC. Immunopositive fibres only sporadically crossed the border between the storage lobe and the glandular lobe of the CC. Immunopositive fibres entered the storage lobe of the CC via the nervus corporis cardiaci I (NCCI); NCCII was immunonegative. Unilateral retrograde fillings of the NCCI with the fluorescent tracer Lucifer yellow, followed by antiserotonin immunocytochemistry, revealed about 20 double-labelled neurones in the anterior part of the pars intercerebralis. The double-labelled neurones were scattered between fluorescent non-immunoreactive neurones. Additionally, 5–7 neurones labelled only with Lucifer yellow were found at the ventrolateral side of the tritocerebrum. No immunopositive neurones were observed in the hypocerebral ganglion. Immunopositive fibres from neurones in the frontal ganglion ran via the recurrent nerve and the neuropile of the hypocerebral ganglion into the paired oesophageal nerve. At most, a few immunopositive nerve fibres occurred in the cardiostomatogastric nerves II, which connect the storage lobe of the CC with the paired oesophageal nerve at the caudal end of the hypocerebral ganglion.
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  • 68
    ISSN: 1432-0878
    Keywords: Vasoactive intestinal peptide ; Substance P ; Enkephalin ; Somatostatin ; Caecum ; Enteric neurons ; Autonomic ganglia ; Immunocytochemistry ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The gross morphology and growth patterns of substance P, enkephalin-, somatostatin and vasoactive intestinal peptide-immunoreactive neurons have been studied in explant cultures of the myenteric plexus taken from beneath the newborn guinea-pig taenia coli, grown for up to 4 weeks in vitro. Substance P and enkephalin-immuno-reactive neurons were more abundant than somatostatin and vasoactive intestinal peptide-immunoreactive neurons. The peptide-containing neuronal cell bodies were clearly visible in culture and exhibited characteristic gross morphologies similar to those described in situ, although some overlap of shape between populations containing different peptides was seen. All four types of peptide-containing fibres were found in the outgrowth and central areas of the cultures. In the case of substance P and somatostatin, the density and pattern of labelling in the central, neuronal area of the cultures resembled that previously seen in the myenteric plexus of the newborn guinea-pig caecum in situ, while the density of the enkephalin-immunoreactive fibres was greater, and that of the vasoactive intestinal peptide-immunoreactive fibres less than that seen in situ. These observations suggest that subpopulations of myenteric neurons containing different peptides may be differentially affected by the culture environment. Possible contributory factors are discussed.
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  • 69
    ISSN: 1432-0878
    Keywords: Octopamine ; Immunocytochemistry ; Nervous system ; Locusta migratoria ; Schistocerca gregaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of octopamine in the metathoracic ganglion, brain and corpus cardiacum of Locusta migratoria and Schistocerca gregaria was investigated by means of immunocytochemistry with an antiserum against octopamine. The dorsal unpaired median (DUM) cells of the metathoracic ganglion were found to be strongly octopamine-immunoreactive. In the rostroventral part of the protocerebrum a group of seven immunopositive cells was demonstrated. Stained nerve fibres of these cells run into three directions: circumoesophageal connectives, midbrain, and optic lobes. As far as the protocerebrum is concerned, immunoreactive fibres were found in the central body, the protocerebral bridge, and in other neuropile areas. In the optic lobe a dense plexus of immunopositive fibres was found in the lobula and in the medulla. In the brain one other immunopositive cell was demonstrated, situated at the lateral border of the tritocerebrum. Octopamine could not be shown to occur either in the globuli cells of the mushroom bodies or in the dorsolateral part of the protocerebrum, where the perikarya of the secretomotor neurones are located that innervate the glandular cells of the corpus cardiacum. In the nervi corporis cardiaci II, which contain the axons of the neurones that extend into the glandular part of the corpus cardiacum, and in the corpus cardiacum proper no specific octopamine immunoreactivity could be found.
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  • 70
    ISSN: 1432-0878
    Keywords: Caudo-dorsal cells ; Neuropeptides ; Immunocytochemistry ; In situ hybridization ; Ovulation hormone ; Lymnaea stagnalis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The caudo-dorsal cells (CDC) in the cerebral ganglia of the pond snail Lymnaea stagnalis synthesize the 36-amino acid ovulation hormone (CDCH). We have used immuno-cytochemistry and in situ hybridization to reveal the localization of neurons and axons containing CDCH-like material. A monoclonal antibody to a fragment of CDCH and a cDNA probe encoding CDCH reacted with the CDC-system, with specific cell groups in the cerebral and pleural ganglia, and with individually occurring neurons throughout the central nervous system. The cells in the pleural ganglia, which were found in about 50% of the preparations studied, are considered as “ectopic” CDC. They are morphologically similar to CDC in their somal dimensions and axonal organization. By means of immuno-electron microscopy it was shown that these neurons contain secretory vesicles that are similar to those of the CDC. The neurons of the bilateral groups occurring in the cerebral ganglia in addition to the CDC are smaller and more intensely stained than the CDC. Axons of these small neurons probably have varicosities located on the CDC axons in the neuropil of the cerebral ganglion, indicating synaptic contacts. Two major axon tracts could be followed from (or toward) the neuropil of the cerebral ganglion. One tract runs from the cerebral gangion via the pleural and parietal ganglia to the visceral ganglion, giving off branches to most nerves emanating from these ganglia. The other tract could be traced through the cerebro-pedal connective to the pedal ganglia. Only in the right pedal ganglion was extensive axonal branching observed. The nerves emanating from this ganglion contained many more immunoreactive axons than those from the left pedal ganglion. A polyclonal antibody raised against the synthetic fragment of CDCH stained, in addition to the neurons and axons revealed with the monoclonal antibody and the cDNA probe, three other major groups of neurons. Two are located in the cerebral ganglion, the other in the left pedal ganglion. The present findings suggest the presence of a system of neurons that contain CDCH or CDCH-like peptides. The role this system may play in the control of egg-laying and egg-laying behaviour is discussed.
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  • 71
    ISSN: 1432-0878
    Keywords: Calcium-binding protein ; Enteric nervous system ; Intestine ; Immunocytochemistry ; Guinea-pig ; Rat ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactivity for vitamin D-dependent calcium-binding protein (CaBP) has been localized in nerve cell bodies and nerve fibres in the gastrointestinal tracts of guinea-pig, rat and man. CaBP immunoreactivity was found in a high proportion of nerve cell bodies of the myenteric plexus, particularly in the small intestine. It was also found in submucous neurons of the small and large intestines. Immunoreactive nerve fibres were numerous in the myenteric ganglia, and were also common in the submucous ganglia and in the intestinal mucosa. Immunoreactive fibres were rare in the circular and longitudinal muscle coats. In the myenteric ganglia of the guinea-pig small intestine the immunoreactivity is restricted to one class of nerve cell bodies, type-II neurons of Dogiel, which display calcium action potentials in their cell bodies. These neurons were also immunoreactive with antibodies to spot 35 protein, a calcium-binding protein from the cerebellum. From the distribution of their terminals and the electrophysiological properties of these neurons it is suggested they might be sensory neurons, or perhaps interneurons. The discovery of CaBP in restricted sub-groups of enteric neurons may provide an important key for the analysis of their functions.
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  • 72
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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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  • 73
    ISSN: 1432-0878
    Keywords: GABA ; Glutamate decarboxylase ; GABA transaminase ; Exocrine pancreas ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural immunohistochemical localization of gamma aminobutyric acid (GABA) and its regulating enzymes, l-glutamate decarboxylase (GAD) and gamma aminobutyrate-α-ketoglutarate transaminase, was determined utilizing an immunogold post-embedding protocol in pancreatic exocrine tissue. Within the acinar cell, GABA and its biosynthetic enzyme, GAD, were localized in zymogen granules. Quantitative analysis of the GABA immunoreactivity in the acinar cell revealed 1.7±0.5 gold particles/μm2 over the cytoplasm, 36.6±14.1 gold particles/ μm2 over the zymogen granules, and 2.9±2.1 gold particles /μm2 over the mitochondria. Quantitative analysis of the distribution of colloidal gold particles, representing glutamate decarboxylase immunoreactivity in the acinar cells, revealed 38.4±2.5 gold particles/μm2 over the zymogen granules, 4.7±1.1 gold particles/μm2 over the mitochondria and 6.3±0.5 gold particles/μm2 over the remainder of the cytoplasm. Substitution of normal sheep serum for the sheep anti-glutamate decarboxylase serum revealed a significant (p〈 0.001) decrease of the colloidal gold particle distribution over the zymogen granules and cytoplasmic compartments of the acini. Gamma aminobutyrate -α-ketoglutarate transaminase, the catabolic enzyme for GABA, was not detected in the mitochondria, zymogen granules, and cytoplasm of the acinar cell, suggesting that GABA is not catabolized within the acinar cell. Preabsorption and substitution controls resulted in an absence of labeling. These results suggest that GABA may act extracellularly and/or have a role within the zymogen granule in the exocrine pancreas.
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  • 74
    ISSN: 1432-0878
    Keywords: MPTP ; Rat ; Nigrostriatum ; Parkinsonism ; Immunocytochemistry ; Tyrosine hydroxylase ; Animal model ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several laboratories have reported that N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine causes damage to the nigral dopamine neurons of man, monkey, and mouse. Controversial data suggest that a rat model of Parkinsonism may be possible. Although loss of dopamine cells has not been detected in the rat brain, our immunocytochemical studies show that immunoreactive tyrosine hydroxylase, the rate-limiting enzyme which synthesizes dopamine, is significantly reduced in concentration, or its antigenicity altered, in substantia nigra/pars compacta as well as the caudate nucleus. Optical density measurements demonstrate the reduction or alteration of immunoreactive tyrosine hydroxylase in nigro-striatal neurons, indicating that axonal terminals, as well as parent perikarya, may be sensitive to the drug. After treatment, abnormal morphological remodelling may result in the affected neuronal processes, perhaps indicating sublethal toxicity, followed by slow recovery. Despite the lack of nigral cell death, it is proposed that the present data support the use of the rat as a model to investigate the early effects of Parkinsonism induced by this agent, and the biological mechanisms of cellular recovery.
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  • 75
    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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  • 76
    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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  • 77
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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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  • 78
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    Cell & tissue research 254 (1988), S. 585-592 
    ISSN: 1432-0878
    Keywords: Epidermis ; Desmosomes ; Immunocytochemistry ; Fine structure ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cultured human epithelial cells stained with antibody to desmosomal proteins by indirect immunofluorescence showed linear arrays of desmosomes en face between stratified cells. To confirm that an extensive linear pattern existed on the cell surface, subconfluent cultures were viewed using scanning electron microscopy. Aligned arrays of blunt protrusions lying parallel to each other and extending in the direction of the long axis of the cell were observed on the surface of groups of superficial cells in intact cultures. That this pattern was indeed related to desmosomal distribution was verified by transmission microscopy of thin sections cut in a plane between the upper and lower surfaces of flattened stratified cells to view desmosomes directly. A similar arrangement of desmosomes was seen in intact tissue, using epidermal sheets separated from newborn foreskin. The same pattern found in flattened cells was sometimes apparent in more rounded basal cells where the cytoplasm was beginning to extend. Since desmosomal plaques are associated with keratin filaments, the alignment of desmosomes must occur in association with cytoskeletal changes as cells become flattened toward the distal epithelial surface. The primary initiation of desmosomal alignment remains to be investigated. However, the present findings demonstrate an increasingly regular membrane-cytoskeletal spatial interaction as stratified epithelial cells of skin mature.
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  • 79
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    Cell & tissue research 254 (1988), S. 635-646 
    ISSN: 1432-0878
    Keywords: Neurotransmitters ; Development, ontogenetic ; Immunocytochemistry ; Nervous system, central ; GABA ; Schistocerca gregaria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of GABA-like immunoreactivity was investigated in embryonic and juvenile locusts using an antibody raised against GABA-protein conjugates. GABA-like immunoreactivity was first detectable in the neuropile of embryonic ganglia at 55% development, and in neuronal somata at 62% development. The total number of immunoreactive somata increased between 62% and 85% embryonic development, and followed an anterio-posterior pattern of expression. At 85% development, the number of immunoreactive somata reached adult levels and no change in number was then seen. In embryonic stages and first and second juvenile instars two dorsal and four ventral groups of somata were labeled in all three thoracic ganglia, whilst in later juvenile instars one of the dorsal groups was visible as a separate entity only in the metathoracic ganglion. These early patterns were modified by alterations in the positions of some of the groups during late embryogenesis and during juvenile development to produce the adult pattern. The results show that the development of GABA expression is similar to that of other neurotransmitters. The characteristics of the development of immunoreactivity indicate that some of these immunoreactive clusters may be derived from clonally related neurones. Finally, we demonstrate the presence of immunoreactive somata and processes in embryos, which correspond to those of identified local and intersegmental interneurones studied in the adult.
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  • 80
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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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  • 81
    ISSN: 1615-6102
    Keywords: Cytoskeleton ; Immunocytochemistry ; Microfilaments ; Microtubules ; Microtubule organizing center ; Uromyces phaseoli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The microtubule and F-actin cytoskeleton of nondifferentiated germlings ofUromyces phaseoli was studied using immunofluorescence methodologies. The microtubules were oriented mostly parallel to the longitudinal axis of the hypha. Microtubule depolymerizing agents, such as cold, demecolcine, griseofulvin and nocodazole, were effective in destroying the microtubule network, but not the F-actin system. Repolymerization of microtubules, following release from these agents, occurred first in the hyphal apices and not near the nuclei or spindle pole bodies. It was concluded that the microtubule nucleating region in such fungal cells is located in the apical regions. Enhanced microtubule arrays were visualized following incubation of the cells in taxol, an agent known to favor microtubule polymerization.
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  • 82
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    Protoplasma 124 (1985), S. 157-167 
    ISSN: 1615-6102
    Keywords: Glyoxysomes ; Gossypium ; Immunocytochemistry ; Isocitrate lyase ; Protein A-gold
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The protein A-gold technique with plastic-embedded sections was used to localize isocitrate lyase (ICL) in cotyledons of mature (50-day-postanthesis) and germinated (24 and 48 hours old) cotton (Gossypium hirsutum) seeds. This is the first report of an ultrastructural localization of a glyoxysomal enzyme by post-embeddment immunocytochemistry. Standard glutaraldehyde, post-osmium tetroxide (3%, 2%) fixation resulted in the retention of more ICL antigenicity in sections of germinated seeds than with aldehyde fixation alone. This became apparent only after overcoming the masking effect of osmium tetroxide on protein antigenicity by pretreatment of sections with sodium metaperiodate. The pattern of gold labeling showed that enzyme was distributed throughout the glyoxysomal matrix including amorphous nucleoid inclusions. Specific association of any enzyme with these structures has not been demonstrated before. Activity of ICL was not detectable spectrophotometrically in extracts or isolated organelles of mature embryos, whereas antigen was demonstrated immunocytochemically and by double immunodiffusion. These results, plus biochemical evidence from our previous work, established the presence of glyoxysomes with a complete complement of enzymes in mature seeds. The increase in ICL activity following germination was accompanied by an increase in density of gold particles over glyoxysomal matrices. Transformation of density values through stereological formulae to a hypothetical number of particles per glyoxysome revealed a 4-fold increase in immunocytochemical detectability which correlated with the 4-fold increase in enzyme activity between 24- and 48-hour post-imbibition. We were unable to detect a site of synthesis despite good ultrastructural preservation of polysomes, likely due to an extremely low concentration of nascent ICL molecules at any polysome when cell metabolism was terminated by fixation. Results of this study support the hypothesis developed in our laboratory that complete glyoxysomes are synthesized during cottonseed maturation, then enlarge and accumulate enzymes by post-translational additions from the cytosol for lipid mobilization during seedling growth.
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  • 83
    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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  • 84
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    Cell & tissue research 186 (1978), S. 551-558 
    ISSN: 1432-0878
    Keywords: C-cells ; Thyroid gland ; Immunocytochemistry ; Calcitonin antibody ; Mammals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the thyroid glands of the horse, pig, deer, mole, and rat, C-cells could be demonstrated by means of the immunocytochemical PAP-technique using rabbit antisera against human calcitonin. Only in ruminants, the crossreaction between the intracellularly stored antigen and the antibodies used appeared to be incomplete.
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  • 85
    ISSN: 1432-0878
    Keywords: Thyroid gland ; Immunocytochemistry ; Thyroglobulin ; Petromyzon marinus L
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies made against thyroglobulin (TG) were used in an immunocytochemical study for the light and electron microscopic localization of TG in the thyroid gland of the anadromous sea lamprey, Petromyzon marinus, during its upstream migration. TG was found in the follicular lumen and in some colloid droplets within the follicular cells. Except for an immunoreactive product observed in a small portion of the interstitial connective tissue, the location of TG in the lamprey was similar to that in the thyroid of the rat.
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  • 86
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Dipnoi (Protopterus) ; Neurohypophysial hormones ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the unlabeled antibody peroxidase-antiperoxidase complex (PAP) technique at the light microscopic level, it has been shown that, in the dipnoan preoptico-hypophysial neurosecretory system, vasotocin and mesotocin are synthesized in separate neurons. In the preoptic nuclei, the perikarya of these two types of neurosecretory neurons are not located preferentially. The two types of neurosecretory perikarya give rise to separate vasotocinergic and mesotocinergic axons, respectively. The dipnoan median eminence and neural lobe contain separate vasotocinergic and mesotocinergic nerve fibres, the general distribution of which is described. In the pars distalis and the pars intermedia of the hypophysis, neurohypophysial hormone-containing nerve fibres have not been found.
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  • 87
    ISSN: 1432-0878
    Keywords: Cyproterone acetate ; d-Norgestrel ; Progesterone ; Pars distalis ; adenohypophysis ; Immunocytochemistry ; Dog (Beagle)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of short-term (8 weeks) treatment with different doses of cyproterone acetate (CPA), d-norgestrel (d-N) and progesterone on cells of the pars distalis, as revealed by the immunoperoxidase technique, were studied in cycle-synchronized beagle bitches (first anoestrus). Pituitary glands from non-treated primiparous beagle bitches at the 6th and 9th week of pregnancy were also included. For immunochemical staining specific antisera to the following hormones were used: canine GH, canine PRL, porcine ACTH, bovine TSHβ, bovine LHβ and human FSHβ. Morphological features of high secretory activity in GH cells were evident even after the human oral contraceptive doses of CPA and d-N, and after a dose as low as 0.1 mg/kg/day subcutaneously (s.c.) of progesterone. In contrast, PRL cells did not show any significant treatment-related effects except in those animals which received the highest dose of d-N (0.5 mg/kg/day per os). In this group, as well as in all pregnant bitches, hyperplasia and hypertrophy of PRL cells were found. In the animals treated with the highest doses of CPA (4.0 mg/kg/day per os) and progesterone (42.5 mg/kg/day s.c.) as well as in pregnant bitches, ACTH/MSH and TSH cells showed marked atrophy and regressive changes. Similar morphological signs of depressed secretory activity were also observed in the cells shown to contain FSHβ and/or LHβ as a result of treatment with the highest dose of progesterone and at the 9th week of pregnancy. These structural responses indicate that quantitative and/or qualitative differences may exist between progesterone, the synthetic progesterone derivative CPA and the nortestosterone type progestagen d-N with regard to their effect on pituitary hormone secretion in the beagle bitch.
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  • 88
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 194 (1978), S. 79-102 
    ISSN: 1432-0878
    Keywords: Duodenum ; Endocrine cells ; Differentiation ; Immunocytochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development and cytodifferentiation of endocrine cells that produce the gastrointestinal hormones gastrin, cholecystokinin and secretin have been studied by a combined fluorescence-cytochemical, immunocytochemical and ultrastructural approach. The results show that, during development, several ultrastructurally distinct cell types exhibit COOH-terminal gastrin and cholecystokinin immunoreactivity. Furthermore, some cells simultaneously contain both gastrin- and cholecystokinin-specific antigenic determinants. Studies on the time course of development of gastrin and cholecystokinin cells, together with the above-mentioned data, suggest that gastrin cells may be converted into cholecystokinin cells in development. During this period, gastrin, cholecystokinin and secretin cells store the biogenic monoamine, 5-hydroxytryptamine a feature not displayed by the adult counter-parts of these cells. In the adult duodenum, characteristic enterochromaffin (EC) cells store 5-hydroxytryptamin for which, evidence for a possible hormonal role has been presented. Taken together, our data indicate that the differentiation of duodenal endocrine cells occurs in distinct steps, each involving a restriction in the biosynthetic repertoire of the cell.
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  • 89
    ISSN: 1432-0878
    Keywords: Pancreas ; Gut ; Pancreatic polypeptide (PP) cells ; Immunocytochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pancreatic polypeptide (PP) cells in the pancreas and gut of chickens were studied by immunocytochemistry. In the pancreas PP cells were numerous and disseminated in the exocrine parenchyma. In this location they were first seen at the 9th day of incubation, i.e. several days after the appearance of glucagon, insulin and somatostatin cells. Very large numbers of these cells occurred from about the 14th day until shortly after hatching when the PP cell frequency was somewhat reduced. At the 17th day of incubation PP cells appeared in the duodenum. Subsequently the number of PP cells in the duodenum increased, and PP cells began to appear also in the jejunum-ileum (19th day) and in the proventriculus and colon (21st day). At hatching and a few days thereafter, PP cells were relatively numerous in the small intestines but much less frequent in the proventriculus and colon. One week after hatching PP cells had disappeared from the colon but remained in the proventriculus and small intestines. Ultrastructurally the PP cell was clearly distinguishable from the insulin, glucagon and somatostatin cells. It was characterized by the presence of spherical cytoplasmic granules which were membrane-bound and moderately electron dense. Areas of firm adhesion between PP cells and acinar cells in the form of desmosomes and possibly also gap junctions were observed.
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  • 90
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Mexican axolotl ; Prolactin ; Somatotropin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunofluorescence procedure was used to identify prolactin (LTH)-and somatotropin (STH)-producing cells in the pituitary of the Mexican axolotl. Histological staining techniques were employed to corroborate immunocytological results. The LTH cells are large, orange-staining cells (acidophils 1) distributed in the posterior two-thirds of the pars distalis. The STH cells are small, erythrosinophilic elements (acidophils 2) principally concentrated in the dorsal part of the pars distalis.
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  • 91
    Electronic Resource
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    Springer
    Cell & tissue research 190 (1978), S. 151-161 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Rhesus monkey ; Immunocytochemistry ; Luteinizing hormone ; Thyroid stimulating hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary glands from juvenile (pre-pubertal) and adult male and female rhesus monkeys were examined following immunocytochemical staining with antisera to the beta subunits of ovine luteinizing hormone (LHβ) and of human thyroid stimulating hormone (TSHβ). The LHβ antiserum reacts with a cell that is PAS-positive, occurs singly and is randomly distributed throughout the pars distalis. The diameter of these cells is approximately 11.5 μm. They do not seem to vary in number in either juveniles (pre-pubertals) or adults, or in males or females. There appears to be fewer LH cells in the pituitary glands of pregnant and lactating females. In addition to staining cells in the pars distalis, the antiserum also reacts with a population of cells located in the pars tuberalis. The cells that stain with the anti-TSHβ serum are confined primarily to the pars distalis. They are approximately 15.8 μm in diameter and are generally found in groups or clusters located in the anterior and medial regions of the gland. The TSH cells vary in number from one animal to another; however, this variability is unrelated to the age or the sex of the animals. No demonstrable changes occur in the number of TSH cells during pregnancy or lactation.
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  • 92
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 193 (1978), S. 211-218 
    ISSN: 1432-0878
    Keywords: 17β-estradiol ; Pituitary gland ; Dog (Beagle) ; Immunocytochemistry ; Glycoprotein hormones (FSH, LH, TSH)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of long-term treatment (52 weeks) with high doses of 17β-estradiol (1.28 mg/kg/week intramuscularly) on gonadotrophs was studied in the pituitary gland of the beagle bitch. For immunochemical staining the immunoperoxidase technique and antisera to the specific beta (β) subunits of FSH and LH were employed. For control purposes antisera to the following hormones were also used: bovine TSHβ, canine GH, canine PRL and porcine ACTH1. In the pars distalis and pars tuberalis of control bitches, in addition to the cells which react solely with antisera to either LHβ or FSHβ, most cells were reactive to both antisera. The cells stained for FSHβ were less numerous than those shown to contain LHβ. TSHβ, PRL, GH and ACTH/MSH were localized in distinctly different cell types in the pars distalis of all control animals. In the treated bitches, almost complete regression of cells classically identified as gonadotrophs and stained for LHβ was observed. On the other hand, using the antiserum to FSHβ, selective immunochemical staining was localized in cells fitting the morphological characteristics of TSH cells. All these cells were also stained for TSHβ. However, a few cells were also shown to react solely with the antiserum to TSHβ. These cells, which seem to contain both TSHβ and FSHβ, were further clearly differentiated from PRL, GH and ACTH/MSH cells on the basis of their cytological features, intraglandular distribution and by immunochemical double staining. These observations support the concept that the one cell-one hormone theory may not necessarily apply to the glycoprotein hormones of the dog pituitary gland.
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  • 93
    Electronic Resource
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    Cell & tissue research 194 (1978), S. 327-336 
    ISSN: 1432-0878
    Keywords: Prolactin ; Growth hormone ; Ovine pituitary ; Immunocytochemistry ; Granule size
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the peroxidase-antiperoxidase immunocytochemical staining technique, prolactin and growth hormone cells have been identified and described in the ovine pituitary. The image analysing computer, Quantimet 720, was used to assess accurately the size range of the secretory granules in these cell types. The area size distributions of the prolactin and growth hormone granules are similar. An increased proportion of larger granules was observed in the prolactin cells post-partum. Serial sections stained alternately for prolactin or growth hormone confirmed that the cells contain either prolactin or growth hormone but not both.
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  • 94
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 195 (1978), S. 183-187 
    ISSN: 1432-0878
    Keywords: Somatostatin-containing neurons ; Hypothalamus ; Median eminence ; Immunocytochemistry ; Domestic mallard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the hypothalamus of the adult domestic mallard, small to medium-sized perikarya are stained specifically with rabbit antiserum against cyclic somatostatin (PAP technique of Sternberger). The somatostatin-immunoreactive material is located in neurons different from those containing immunoreactive LHRH, vasotocin or mesotocin. Somatostatin-containing perikarya are observed 1) in a chain-like arrangement extending from the area of the median division of the supraoptic nucleus to the caudal end of the paraventricular nucleus; 2) as single cells in the preoptic region; and 3) as a conspicuous formation in the optic tract division of the supraoptic nucleus. In the rostral portion of the median eminence, somatostatin-immunoreactive axons penetrate into the external zone. Fine accessory fiber bundles project to the neural lobe.
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  • 95
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 186 (1978), S. 393-398 
    ISSN: 1432-0878
    Keywords: β-Lipotropin ; Hypothalamus ; Hypophysial portal capillaries ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary β-Lipotropin (β-LPH) has been localized in hypothalamus and pituitary of sheep and ox by the immunoperoxidase technique. In both species β-LPH was found in perikarya of arcuate neurons as well as in cells of the anterior and intermediate lobes of the pituitary. A large number of immunoreactive axons were found in the arcuate region; some appeared to innervate other neurons and others projected to portal capillaries. Stained fiber segments were also scattered throughout the hypothalamus. The presence of β-LPH in hypothalamic neurons supports the possibility that brain β-LPH may be a precursor for opiate-like or other peptides which may be involved in neuromodulation or neurohormonal activities.
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