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  • Immunocytochemistry  (43)
  • Springer  (43)
  • Annual Reviews
  • Blackwell Publishing Ltd
  • National Academy of Sciences
  • 2005-2009
  • 1990-1994  (43)
  • 1980-1984
  • 1940-1944
  • 1990  (43)
Collection
Publisher
  • Springer  (43)
  • Annual Reviews
  • Blackwell Publishing Ltd
  • National Academy of Sciences
  • Wiley-Blackwell  (3)
Years
  • 2005-2009
  • 1990-1994  (43)
  • 1980-1984
  • 1940-1944
Year
  • 1
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Met-enkephalin-Arg6-Phe7 ; Met-enkephalin-Arg6-Gly7-Leu8 ; Neurosecretory cells of insects ; Neuropeptides ; Co-existence of peptides ; Blowfly,Calliphora vomitoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuronal pathways in the retrocerebral complex and thoracico-abdominal ganglionic mass of the blowflyCalliphora vomitoria have been identified immunocytochemically with antisera against the extended-enkephalins, Met-enkephalin-Arg6-Phe7 (Met-7) and Met-enkephalin-Arg6-Gly7-Leu8 (Met-8). Neurons of the hypocerebral ganglion, immunoreactive to Met-8, have axons in the crop duct nerve and terminals in muscles of the crop and its duct. Certain neurons of the hypocerebral ganglion are also immunoreactive to Met-7, and axons from these cells innervate the heart. Met-8 immunoreactive nerve terminals invest the cells of the corpus allatum. The source of this material is believed to ve a single pair of lateral neurosecretory cells in the brain. There is no Met-7 immunoreactive material in the corpus allatum. In the corpus cardiacum neither Met-7 nor Met-8 immunoreactivity is present in the cells. However, in the neuropil of the gland certain fibres, with their origins elsewhere, do contain Met-8 immunoreactivity. The most prominent neurons in the thoracic ganglion are the Met-7 immunoreactive ventral thoracic neurosecretory cells, axons from which project to neurohaemal areas in the dorsal neural sheath and also, via the ventral connective, to the brain. Co-localisation studies show that the perikarya of these cells are immunoreactive to antisera raised against several vertebrate-type peptides, such as Met-7, gastrin/cholecystokinin and pancreatic polypeptide. However, their axons and terminals show varying amounts of the peptides, suggesting differential transport and utilisation. Only a few cells in the thoracic ganglion are immunoreactive to Met-8 antisera. These lie close to the nerve bundles suppling the legs. In the abdominal ganglion, Met-8 immunoreactive neurons project to the muscles of the hindgut. This study suggests that the extended enkephalin-like peptides ofCalliphora may have a variety of different roles: as neurotransmitter or neuromodulator substances; in the direct innervation of effector organs; and as neurohormones.
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  • 2
    ISSN: 1432-0878
    Keywords: Insulin-like peptide ; Immunocytochemistry ; Immunochemical characterization ; Brain ; Neuroendocrine structures ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical tests with eight monoclonal antibodies against either bovine or human insulin and seven polyclonal antibodies against bovine insulin were carried out to determine the presence of insulin-like neuropeptides in the brain and affiliated neuroendocrine structures of the insect Leucophaea maderae. Reaction products identified in the brain, subesophageal ganglion, and corpus cardiacum-corpus allatum complex indicate the presence of materials resembling mammalian insulins in its antigenic properties. The immunostaining observed with monoclonal antibodies appears to indicate the occurrence of an insulin-related peptide that shows sequential similarities with parts of both the A- and B-chains of mammalian insulin molecules. These suppositions are supported by the results of dot-blot and two-site time-resolved immunofluorometric assay (TR-IFMA) screenings of fractions of Leucophaea tissue extracts obtained by chromatography. The polyclonal antibodies yielded reaction products in some of the same areas and in additional parts of the neuroendocrine system not visualized by the monoclonal antibodies. Immunoreaction was observed in the following areas: the pars intercerebralis of the protocerebrum, the nervi corporis cardiaci I transporting insulin-like material to the corpus cardiacum, the dorsolateral protocerebral area and the optic lobes, the deutocerebrum, the tritocerebrum, and the subesophageal ganglion. In addition, smaller cell bodies with immunoreactive deposits occur at the border between proto- and deutocerebrum, and in the central area of the protocerebrum. The distribution of reactive material in the corpus cardiacum-corpus allatum complex after use of both groups of antibodies was the same. The fact that polyclonal and monoclonal antibodies yielded reaction products in different cells of the brain suggests either that the two groups of antibodies recognize different epitopes of the same molecule, or that they reveal two different types of immunoreactive molecules related to mammalian insulins. Together with the biochemical data reported by Nagasawa and coworkers (PNAS 83, 1986) the present immunocytochemical analysis has established a closer relationship between mammalian and insect “insulins” than was previously known.
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  • 3
    ISSN: 1432-0878
    Keywords: Serotonin ; Urotensins ; Somatostatin ; Immunocytochemistry ; Caudal neurosecretory system ; Reissner's fiber (subcommissural organ) ; Salmon,Oncorhynchus kisutch (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The caudal spinal cord of the coho salmon was investigated by means of immunocytochemistry using antisera against serotonin, urotensin I, urotensin II, somatostatin and a urea-extract of bovine Reissner's fiber (AFRU). Populations of serotonin-immunoreactive (IR) neurons were found rostral and dorsal to the urophysis in close spatial association with caudal secretory neurons. Thick, smooth serotonin-IR processes extended toward the external surface of the spinal cord where they displayed conspicuous terminal dilatations. Thin, beaded serotonin-IR fibers appeared to innervate populations of caudal secretory and somatostatin-IR cerebrospinal fluid-contacting neurons. Most caudal neurosecretory cells displayed both urotensin I and urotensin II immunoreactivities; only a minority reacted exclusively with either urotensin I or urotensin II antisera. Urotensin II-IR and somatostatin-IR cerebrospinal fluid (CSF)-contacting neurons were found as an integral component of the central canal wall in the caudal spinal cord and filum terminale; their dendritic processes appeared to contact Reissner's fiber, which displayed a weak AFRU-immunoreactivity while inside the central canal, but became strongly reactive in the interior of the terminal ventricle as it formed the massa caudalis. The distribution of serotoninergic processes points to a regulatory role in the function of caudal secretory and CSF-contacting neurons and to a putative serotonin release into the subarachnoid space and/or meningeal vasculature. It is also suggested that the CSF-contacting neurons of the central canal may participate in a feedback mechanism controlling the secretory activity of the subcommissural organ.
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  • 4
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Pars tuberalis ; Immunocytochemistry ; Thyroid-stimulating hormone (TSH) ; Propylthiouracil (PTU) ; Thyroxine (T4) ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pars tuberalis (pt) of the adenohypophysis is unique in its close spatial relationship to the neurohemal contact area of the median eminence. The morphology of pt-specific secretory cells does not resemble cell types of the pars distalis (pd); the functional role of these cells within the endocrine system is still unknown. One group of young mature female Wistar rats received propylthiouracil (PTU), a second group thyroxine (T4) (10 mg/l each in drinking water) from about 3 weeks prior to the expected pregnancy and throughout the experiment. On gestation day 20, the fetuses were obtained by laparatomy. Serial sections from the rostral portion of the pt and from the pd were immunostained using the peroxidase-antiperoxidase method. TSH concentrations were determined by RIA in serum and pituitaries; T4 was measured in serum. An antiserum against rat (r) TSH revealed a moderate positive reaction of nearly all cells of the pt in the control group. In both experimental groups the pt-specific cells showed weak or no immunoreactivity. Sections of all groups were negative with anti(r)-LH,-GH,-PRL. In contrast to controls, only a few immature TSH-cells could be found in sections of the pd in the T4-group, while concentrations of TSH in blood and hypophysis were very low. TSH-cells in the PTU-group were enlarged and less intensely stained. TSH-concentrations were decreased in the hypophysis, blood levels were elevated. All sections of the pd-specific cell populations showed positive immunoreactions with anti(r)-LH,-GH,-PRL. The present results suggest that pt-specific secretory cells of the fetal rat possess TSH immunoreactivity but do not resemble the thyrotropes of the pd. Marked differences in immunoreactivity displayed by the experimental groups indicate that pt-specific cells respond to changes in the fetal thyroid status and are a component of the thyroid-regulating system in addition to the thyrotropes of the pd. This novel aspect of pt function is discussed in connection with recent results concerning melatonin receptors found in the pt and the inhibitory influence of the pineal gland exerted on the thyroid gland.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 260 (1990), S. 479-484 
    ISSN: 1432-0878
    Keywords: Histamine ; Immunocytochemistry ; Nervous system ; Excretory system ; Flatworms (Platyhelminthes) ; Diphyllobothrium dendriticum (Cestoda) ; Microstomum lineare (Turbellaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific antibodies against histamine were used to demonstrate the occurrence and cellular distribution of histamine-like immunoreactivity in three species of flatworms (phylum Platyhelminthes). In the parasitic cestode Diphyllobothrium dendriticum, histamine-reactivity was found in neurons of the main nerve cords, and in cells lining the central and peripheral excretory ducts. In the free-living microturbellarian Microstomum lineare and in the planarian Polycelis nigra, histamine-immuno-reactivity was restricted to cells and fibres of the nervous system. The occurrence of histamine or a related substance in the nervous system of flatworms, which represent primary bilateria, indicates the importance of this neuroactive substance in the animal kingdom.
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  • 6
    ISSN: 1432-0878
    Keywords: Fundic mucosa ; Stomach ; Pepsinogen ; Cell renewal ; Development, ontogenetic ; Immunocytochemistry ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Development and maturation of pepsinogen 1-producing cells were studied in the gastric fundic mucosa of the mouse by means of light- and electron-microscopic immunocytochemistry using rabbit anti-rat pepsinogen 1-serum. In the adult mouse, secretory granules in mucous neck cells, transitional mucous neck/chief cells and chief cells are immunolabeled. The numerical density of gold particles on zymogen granules is not significantly altered among different stages of maturation of chief cells. In addition, rough endoplasmic reticulum and Golgi complex of these cell types show a weak labeling. In mice from day 16 of gestation to postnatal day 14 mucous neck cells and chief cells cannot be distinguished, but only one type of pepsinogen 1-producing cell, called ‘primitive chief cell’, is identified in the fundic gland. The intensity of immunoreactivity of secretory granules in primitive chief cells is uniform within an individual cell but varies greatly among different cells. The majority of primitive chief cells contains weakly labeled granules regardless of the maturation stage of cells or of animals. On postnatal day 21, mucous neck, transitional and chief cells are distinguishable, and secretory granules in these cells are intensely immunolabeled as in the adult. These results suggest that pepsinogen 1-production rapidly increases with differentiation of mucouse neck and chief cells.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 260 (1990), S. 203-205 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Prolactin cells ; Pituitary gland ; Tilapia larvae, Oreochromis mossambicus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using an antiserum to highly purified chum salmon prolactin, prolactin cells were identified in the putative rostral pars distalis of newly hatched tilapia larvae (Oreochromis mossambicus) by the immunogold method for the electron microscope. In the putative rostral pars distalis, some cells had another kind of secretory granule which was much less numerous, much smaller in size, and without immunoreactivity to salmon prolactin antiserum. Controls incubated with salmon prolactin-preabsorbed antiserum or normal serum showed no immunoreactive cells, confirming the specificity of the antiserum. The possible role of prolactin in the osmoregulation of tilapia larvae is discussed.
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  • 8
    ISSN: 1432-0878
    Keywords: Antennae ; Motoneurons ; Immunocytochemistry ; Cobalt labelling ; GABA ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In crickets, a deutocerebral motoneuron sends axon collaterals to 6 of the 7 antennal muscles. Previous results indicated that this neuron exerts inhibition on these muscles and thus may be a common inhibitory motoneuron. In our present study, we show by doublelabelling, i.e. retrograde cobalt-filling and GABA-immunocytochemistry, that this neuron is GABA-immunoreactive, thus demonstrating that one common inhibitory motoneuron is part of the antennal motor system of crickets.
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  • 9
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Immunocytochemistry ; Submucosal plexuses ; Enteric nervous system ; Small intestine ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In addition to differences between the two submucosal ganglionic neural networks, i.e., the plexus submucosus externus (Schabadasch) and the plexus submucosus internus (Meissner), with respect to the occurrence and distribution of serotonin as neurotransmitter, immunocytochemistry also revealed a distinct distribution for various neuropeptides in these two plexuses. Immunoreactivity for galanin, vasoactive intestinal polypeptide, calcitonin gene-related peptide, substance P, neuromedin U, enkephalin, somatostatin and neuropeptide Y was found in varicose and non-varicose nerve fibres of both submucosal ganglionic plexuses, albeit with a distinct distributional pattern. The difference in neurotransmitter and/or neuromodulator content between both neural networks became even more obvious when attention was focussed on the immunoreactivity of the nerve cell bodies for these substances. Indeed, neuropeptide Y, enkephalin-and somatostatin-immunoreactive neuronal perikarya as well as serotonergic neuronal cell bodies appear solely in the plexus submucosus externus. Neuromedin U-immunoreactive perikarya, mostly coexisting with substance P, are observed in large numbers in the plexus submucosus internus, whilst they are rare in the plexus submucosus externus. Double-labelling immunostaining for substance P with CGRP and galanin revealed a different coexistence pattern for the two submucosal ganglionic plexuses. The differing chemical content of the neuronal populations supports the hypothesis that the existence of the two submucosal ganglionic plexuses, present in most large mammals including man, not only reflects a morphological difference but also points to differentiated functions.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 262 (1990), S. 177-190 
    ISSN: 1432-0878
    Keywords: FMRFamide ; Neuropeptide ; Immunocytochemistry ; Nervous system, central ; Neurohormones ; Helix pomatia (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of FMRFamide-like immunoreactive (FLI) neurons and their morphological characteristics have been investigated in the central nervous system of the snail, Helix pomatia L. Approximately phageal ganglion complex. More than 50% of the FLI neurons were located in the cerebral ganglia. The FLI neurons could be divided into four groups according to size: (i) giant neurons (over 100 μm); (ii) large neurons (80–100 μm); (iii) medium-sized neurons (40–70 μm); (iv) small neurons (12–30 μm). They were distributed i) in groups or clusters, typical of small neurons and ii) in solitary form or in groups comprising 2–3 cells, typical of large and giant neurons. Giant and large neurons revealed only limited arborizations in the neuropil, but rich branching towards and in the peripheral nerves. Some of the small neurons had extensive arborizations of varicose fibers in the neuropil. They may therefore play some role in integratory processes. Varicose FLI fibers were visualized in the cell body layer of the different ganglia, and in the neural sheath of both the ganglia and the peripheral nerves. We propose a multifunctional involvement of FLI neurons and FMRFamide-like neuropeptides in the Helix nervous system: (i) a synaptic or modulatory role in axo-axonic interactions in the neuropil; (ii) a direct influence on neuronal cell bodies in the cortical layer, (iii) innervation of different peripheral organs; and (iv) remote neurohormonal control of peripheral events through the neural sheath.
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  • 11
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Gonadotropes ; Morphometry ; Stereology ; Rana pipiens (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous physiological results have indicated the existence of two releasable pools of gonadotropins in amphibian pituitaries: an acute releasable pool that appears independent of protein synthesis, and a storage pool involved in chronic release that depends on protein synthesis. To elucidate the ultrastructural localization of these pools and the morphological changes induced in gonadotrope cells after treatment with gonadotropin-releasing hormone, we carried out a morphometric study of immuno-identified gonadotrope cells using an in vitro superfusion system. Treatment with gonadotropin-releasing hormone induced a degranulation of small (110–255 nm) and medium (236–360 nm) secretory granules as well as hypertrophy of the endoplasmic reticulum and Golgi complex. Simultaneous incubation with gonadotropin-releasing hormone and cycloheximide inhibited the release of secretory granules although the endoplasmic reticulum and Golgi complex were hypertrophied. These morphological results strongly suggest: (1) that gonadotropin-releasing hormone induces degranulation and hypertrophy of the biosynthetic machinery in gonadotrope cells; and (2) that the activation of the endoplasmic reticulum and Golgi complex by stimulation with gonadotropin-releasing hormone is independent of protein synthesis, while the release of secretory granules is protein synthesis-dependent. In addition, the second or “storage” pool of gonadotropin is associated mainly with the small and medium secretory granules.
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  • 12
    ISSN: 1432-0878
    Keywords: Chang hepatoma cells ; Growth hormone ; GERL ; Golgi complex ; Immunocytochemistry ; Tumor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The binding and internalization of endogenous growth hormone in Chang hepatoma cells were localized on the cell surface and in the Golgi-endoplasmic reticulum-lysosome (GERL) area by various indirect immunocytochemical labeling techniques, namely, peroxidase or colloidal gold conjugated to secondary antibody, and avidin-biotin complex methods. Rabbit antiserum and monoclonal antibodies raised against HPLC-purified porcine growth hormone were used in this study. In fixed material, antigen-antibody complexes were found to be homogeneously distributed along the cell membrane. Control groups showed negative binding on the cell surface. Trypsin treatment before immunolabeling removed antibody binding completely, but hyaluronidase was ineffective. Pretreatment of lectins did not block the recognition of primary antibody to antigen molecules on cell surface. Internalization of the antigen-antibody peroxidase or gold complexes was demonstrated in the cells, which were immunolabeled at 4°C, and then reincubated for 0–30 min at 37°C before fixation. After reincubation, the internalized ligand complexes were found in vesicles near the cell surface or in the GERL area near the Golgi apparatus which, however, did not label for peroxidase. These findings suggest that the trypsin-sensitive growth hormone, specifically bound and internalized into Chang hepatoma cells, is localized in the GERL instead of the Golgi apparatus and might be involved in the mechanism of tumor cell growth.
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  • 13
    ISSN: 1432-0878
    Keywords: Galanin ; Immunocytochemistry ; Necturus maculosus (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Galanin is a biologically active peptide which has a wide pattern of distribution in the mammalian central and peripheral nervous systems. However, the distribution of galanin-like immunoreactivity in amphibian species has not been well elucidated. In the present study, biochemical and immunohistochemical techniques were used to determine the relative concentrations, biochemical nature, and cellular localization of galanin-like immunoreactivity in the brain, heart, urinary bladder, and small intestine of Necturus maculosus (common name: mudpuppy). The results of this study indicate that each of these types of tissue contain a galanin-like peptide which is similar to porcine galanin. Brain and heart concentrations of galanin-like immuno-reactivity were particularly high, although substantial amounts were also present in the small intestine and urinary bladder. Galanin immunoreactivity was observed in ascending fiber tracts of the brainstem and in fibers in the hypothalamus. In addition, galanin immunoreactivity was observed in autonomic neurons and processes in the heart, bladder, and small intestine. The pattern of distribution of galanin-like immunoreactivity in many tissues of this amphibian species is similar to the previously described mammalian pattern; however, galanin-immunoreactive innervation of cardiac tissue has not been reported in mammals. We suggest that galanin-like immunoreactivity in the heart may be more extensive in amphibian species than in mammals.
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  • 14
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Photoreceptor cells ; Rhodopsin ; Membrane recycling ; Cherax destructor (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Examination of the ultrastructure of retinula cells of the Australian crayfish Cherax destructor at different times over a 24-hour cycle, together with patterns of anti-rhodopsin antigenicity, has lead to the formulation of a model of photoreceptor membrane turnover in these animals. Its main features are: (a) the existence of two bursts of rhabdomeral membrane breakdown; one, light-sensitive and synchronous, occurring at dawn, the other, constituting the first part of the membrane replacement phase itself, occurring during the afternoon and night, (b) the desynchronisation of the replacement phase of turnover between animals and to a lesser extent between cells of the same retina, (c) confinement of ultrastructurally detectable signs of photoreceptor membrane processing to the retinula cells themselves, and (d) replacement of a substantial part if not all of the rhabdomeral membrane daily. This model is compatible with many of the observations reported on the American crayfish Procambarus, and utilises the same basic mechanisms that are believed to operate in photoreceptor membrane turnover in many other arthropod compound eyes.
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  • 15
    ISSN: 1432-0878
    Keywords: Pinealocytes ; Visinin ; Calcium-binding protein ; Light, constant ; Photosensitization ; Immunocytochemistry ; Domestic fowl (Gallus domesticus)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Visinin, a calcium-binding protein isolated from the soluble fraction of homogenized chick retinae, has been recognized immunocytochemically in the pinealocytes of various submammals. In the chick pineal organ, continuous environmental light induced an increase in population density of visinin-immunoreactive pinealocytes. In semi-quantitative, dot-immunoblotting analysis, the amount of visinin in the pineal organs of chicks kept under continuous light for 3 days was 4–8 fold more abundant than that under continuous darkness for the same duration. Eye-enucleation and organ culture experiments clarified that this lighting effect was exerted directly on the pineal organ through the skull, and not via the neural pathway including the retinohypothalamic projection. These data suggest the existence of direct photosensitivity in the chick pinealocyte itself and the possible involvement of visinin in photoreception of the pineal organ as well as the retina of chicks.
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  • 16
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasotocin ; Hypothalamus ; Neurosecretory fibers ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of vasotocin-like peptides in the central nervous system of the cartilaginous fish Scyliorhinus canicula was determined by indirect immunofluorescence and peroxidase anti-peroxidase techniques, using a specific antiserum raised in rabbits against synthetic vasotocin. Immunoreactive perikarya were mainly detected in the anterior hypothalamus, within the midcaudal part of the preoptic nucleus. The most rostral positive cell bodies were located in the dorso-lateral parts of the preoptic area, whereas at a more caudal level, they took a ventro-medial position within the deepest layers of the nucleus. Throughout the preoptic region these cells varied in shape according to their location. Occasionally, scattered vasotocin-like immunopositive cells were also identified in the nucleus periventricularis hypothalami. Vasotocin immunoreactivity was detected in numerous varicose nerve fibers of the preopticohypophysial tract. These fibers were seen to course through the medio-basal hypothalamus and caudally, after having passed the hypophysial stem, they reached the neurointermediate lobe of the pituitary. Numerous immunoreactive fibers were also observed within the rostro-medial region of the median eminence. At this level the fibers were in close proximity to the capillary loops. In the preoptic region, some stained cells exibited short processes that appeared to contact non-reactive perikarya. By comparing the distribution of vasotocin- and corticotropin-releasing factor immunoreactivity on adjacent then serial sections, it was revealed that these peptides, in S. canicula, do not coexist in the same perikarya. The present results, are compared with those obtained in other vertebrate groups, and their possible functional implications are discussed.
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  • 17
    Electronic Resource
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    Cell & tissue research 262 (1990), S. 543-550 
    ISSN: 1432-0878
    Keywords: PYY ; NPY ; CGRP ; Serotonin ; Lung ; Radioimmunoassay ; Immunocytochemistry ; Mesocricetus auratus (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We investigated the presence of peptide YY, neuropeptide Y, calcitonin gene-related peptide and serotonin in the hamster lung by radioimmunoassay, high performance liquid chromatography and immunocytochemistry. Lung-tissue concentrations of peptide YY and neuropeptide Y were 1.3±0.2 and 2.5±0.2 pmol/g wet weight, respectively. These two closely related pancreatic peptides were demonstrated in separate peaks with high performance liquid chromatography. The peptide YY appeared fragmented as immunoreactive peptide YY eluted primarily late in the gradient but showed additional peaks early in the gradient. Peptide YY-like immunoreactivity (PYY-LI) was predominantly observed in one or more cells of neuroepithelial bodies in all airways peripheral to bronchioles, and in solitary neuroendocrine cells primarily located in the same peripheral areas. Neuropeptide Y-LI was seen in individual, thin nerve fibers around arteries and veins, in the airway lamina propria, and in the airway epithelium; in the latter also immunopositive nerve terminals were located. This pattern did not appear to coincide with that of calcitonin gene-related peptide-LI in epithelial nerve fibers and terminals. Peptide YY-LI, calcitonin gene-related-LI and serotonin-LI were present in cells of one and the same neuroepithelial body. However, peptide YY-LI was never found to be co-localized with calcitonin gene-related-LI or serotonin-LI, but the latter two were co-localized as previously reported.
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  • 18
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Neurosecretion ; Vasotocin-neurophysin precursor ; Immunocytochemistry ; Lectin histochemistry ; Snake, Natrix maura ; Lizard, Liolaemus cyanogaster
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The immunocytochemical and lectin-binding properties of the magnocellular neurosecretory neurons in the hypothalamus of 2 reptilian species, the snake Natrix maura and the lizard Liolaemus cyanogaster, were investigated. Particular attention was paid to the secretory droplets present in these neurons. Antisera against bovine neurophysins I+II, arginine-vasotocin, and mesotocin were used. The following lectins were applied: concanavalin A (Con A), wheat-germ agglutinin (WGA), and Limax flavus agglutinin (LFA). Adjacent 1-μm-thick methacrylate sections were used to investigate the same secretory neuron and the same colloid droplets with all three antisera and all three lectins. Several sections were treated with trypsin and urea before immunostaining or lectin binding. Con A bound to both vasotocin- and mesotocin-immunoreactive neurons, WGA exclusively to vasotocin neurons; neither of these neurons reacted with LFA. The colloid droplets were present in vasotocin neurons but absent in the mesotocin neurons. These secretory droplets showed an affinity for Con A but not for WGA, and reacted with antisera against neurophysins and vasotocin. In Natrix maura, the colloid droplets became reactive with Con A and the antisera used only after pretreatment of the sections with trypsin and urea. Within the hypothalamo-neurohypophyseal system, antiserum against vasotocin and WGA revealed the same fiber bundles. It is concluded (i) that in reptiles the vasotocin-neurophysin precursor is glycosylated, (ii) that vasotocin neurons have the exclusive capacity to form colloid droplets, and (iii) that these droplets are an intracisternal (RER) storage form of the vasotocin-neurophysin precursor.
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  • 19
    ISSN: 1432-0878
    Keywords: Thyroid ; Neuromedin U ; C-cell ; Immunocytochemistry ; Chromatography ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neuromedin U is a novel neuropeptide found to have a widespread distribution extending throughout the mammalian central nervous system, gastrointestinal tract and the endocrine cells of the pituitary gland. In order to investigate the possibility that neuromedin U-like immunoreactivity is also present in the thyroid gland of the adult rat we have examined its localisation and molecular nature by radioimmunoassay, immunocytochemistry and chromatographic analysis. The neuromedin U content of the whole thyroid gland was found to be 331±67 fmol/gland (mean±SEM), and this value significantly decreased (163±17 fmol/gland) as a result of 14 days of treatment with the anti-thyroid agent methimazole (10 mg/rat/day. Thyrotoxicosis induced by exogenous T4 (10 μg/rat/day) failed to alter the thyroid content of this peptide. Immunostaining studies localised neuromedin U to a minor population of parafollicular C-cells in untreated animals. Complementary chromatographic studies revealed a single molecular form of neuromedin U-like immunoreactivity in thyroid tissue extracts which was indistinguishable from synthetic rat neuromedin U standard.
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  • 20
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    Cell & tissue research 260 (1990), S. 41-48 
    ISSN: 1432-0878
    Keywords: Keratin filament ; Circumvallate papilla ; Taste bud ; Immunocytochemistry ; Electron microscopy ; Mouse (dd)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Keratin filaments of epithelial- and taste-bud cells in the circumvallate papillae of adult and developing mice were studied by immunocytochemistry using monoclonal antikeratin antibodies (PKK2 and PKK3) and by conventional electron microscopy. Elongated cells (type-I,-II, and-III cells) of the taste buds were stained by PKK3 antibody, which reacts with 45-kdalton keratin, whereas basal cells of the taste buds and surrounding epithelial cells showed negative staining with PKK3. Such PKK3-reactive cells occurred at 0 day after birth, when taste-buds first appeared in the dorsal surface epithelium of the papillae. Thus 45-kdalton keratin seems to be an excellent immunocytochemical marker for identifying taste-bud cells. Epithelial cells in all layers of the trench wall and basal layer cells of the dorsal surface contained densely aggregated bundles of keratin filaments that reacted with PKK2 antibody, but not with PKK3. In contrast, taste-bud cells and spinous and granular layer cells of the dorsal surface possessed loose aggregated bundles of filaments that reacted with PKK3, but not with PKK2. These results suggest that the aggregation and distribution pattern of keratin filaments may reflect differences in the keratin subtypes that comprise these filaments.
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  • 21
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    Cell & tissue research 261 (1990), S. 73-84 
    ISSN: 1432-0878
    Keywords: Muscle, skeletal ; Regeneration ; Thrombospondin ; Fibrinogen ; Injury ; Immunocytochemistry ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Biochemical and immuno-microscopic techniques were used to study temporal involvement of thrombospondin in relation to fibrinogen in muscle regeneration using a rat skeletal muscle-wound model. In undamaged control muscle, no fibrinogen and minimal thrombospondin antigen was found. Following crushing injury, fibrin networks appear immediately, followed by a gradual ordered accumulation of thrombospondin (within a few hours) in the vicinity of the vascular bed and adjacent endomysial connective tissue. Later, thrombospondin becomes associated with connective tissue and basal laminae around muscle fibers throughout the damaged muscle, maximal labelling occurring 3–6 days post-injury. Thrombospondin immunoreactivity decreased thereafter to near normal levels after 7 days post-injury, coincident with the appearance of regenerating muscle fibers. In contrast, little fibrin material remained by five days after injury. Quantitative radioimmunoassay of soluble thrombospondin antigen and radioimmune labelling of thick frozen sections reinforced the qualitative immuno-microscopic observations, with levels peaking at 3–4 days post-trauma, 10-fold over control levels. SDS-PAGE immunoblotting of non-reduced muscle extracts three days after a crush assault shows that the bulk of the thrombospondin incorporated into the injury site exists in a polymerized state (≤1000 kD). These results demonstrate that the temporal appearance and disappearance of thrombospondin in the healing of a crushing lesion in muscle is related more closely to the regeneration phase of muscle than to the coagulation phase.
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  • 22
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    Cell & tissue research 261 (1990), S. 107-113 
    ISSN: 1432-0878
    Keywords: Dynorphin A ; GABA ; Substantia nigra ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The opioid peptide dynorphin A (1–17) is the third transmitter identified in the striatonigral projection, the other two being gamma-aminobutyric acid (GABA) and substance P. The ultrastructural features of the dynorphinergic terminals in substantia nigra/pars reticulata were studied using pre-embedding immunocytochemistry with the classical peroxidase-antiperoxidase-diaminobenzidine-method; these features were compared with GABAergic boutons visualized with an immunogold method. Two distinct types of dynorphin-A-immunoreactive boutons could be identified: (1) type A (81%) possessing characteristics similar to the GA-BAergic nerve endings in this region, i.e., large pleomorphic vesicles and symmetric synaptic contacts, (2) type B (19%) displaying asymmetric synaptic zones and small, mostly round vesicles. These results are in agreement with physiological studies suggesting a dual action of dynorphin A in substantia nigra.
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  • 23
    ISSN: 1432-0878
    Keywords: Neuropeptides ; FMRFamide ; Immunocytochemistry ; Insect nervous system ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using an antiserum against the tetrapeptide FMRFamide, we have studied the distribution of FMRFamide-like substances in the brain and suboesophageal ganglion of the sphinx mothManduca sexta. More than 2000 neurons per hemisphere exhibit FMRFamide-like immunoreactivity. Most of these cells reside within the optic lobe. Particular types of FMRFamide-immunoreactive neurons can be identified. Among these are neurosecretory cells, putatively centrifugal neurons of the optic lobe, local interneurons of the antennal lobe, mushroom-body Kenyon cells, and small-field neurons of the central complex. In the suboesophageal ganglion, groups of ventral midline neurons exhibit FMRFamide-like immunoreactivity. Some of these cells have axons in the maxillary nerves and apparently give rise to FMRFamide-immunoreactive terminals in the sheath of the suboesophageal ganglion and the maxillary nerves. In local interneurons of the antennal lobe and a particular group of protocerebral neurons, FMRFamide-like immunoreactivity is colocalized with GABA-like immunoreactivity. This suggests that FMRFamide-like peptides may be cotransmitters of these putatively GABAergic interneurons. All FMRFamide-immunoreactive neurons are, furthermore, immunoreactive with an antiserum against bovine pancreatic polypeptide, and the vast majority is also immunoreactive with an antibody against the molluscan small cardioactive peptide SCPB. Therefore, it is possible that more than one peptide is localized within many FMRFamide-immunoreactive neurons. The results suggest that FMRFamide-related peptides are widespread within the nervous system ofM. sexta and might function as neurohormones and neurotransmitters in a variety of neuronal cell types.
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  • 24
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    Cell & tissue research 261 (1990), S. 367-373 
    ISSN: 1432-0878
    Keywords: Angiotensinogen ; Ovary ; Estrous cycle Renin-angiotensin system ; Atresia ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study examined the presence and cellular distribution of angiotensinogen, the precursor to the angiotensin peptides, in the ovary of the normal cycling rat by immunocytochemistry. Angiotensinogen staining was present in the granulosa cells of maturing follicles and to a lesser extent in those undergoing atresia. Staining was not seen in the granulosa cells of primordial or early primary follicles. In maturing follicles intense staining for angiotensinogen was confined to the antral cell layers, cells of the cumulus oophorus and in the follicular fluid. Strong immunostaining was also seen in the germinal epithelium covering the ovary. Lighter angiotensinogen staining was observed in some parts of the cortical and medullary stroma and occasionally in corpora lutea. No variation in the intensity or pattern of angiotensinogen staining was observed throughout the estrous cycle. Comparison of the distribution of angiotensinogen with the previously described localization of renin, AII, angiotensin converting enzyme and AII receptors, suggests that there are a number of intra-ovarian sites at which AII could be produced.
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  • 25
    ISSN: 1432-0878
    Keywords: Chorionic gonadotropin ; Placental lactogen Placenta ; Cryoultrastructure ; Immunocytochemistry ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The sites of intracellular synthesis and storage of human placental lactogen (hPL) and human chorionic gonadotropin (hCG) are controversial. We have used one of the most sensitive methods, cryoultramicrotomy and immunogold labelling, to localise these hormones at the electron-microscopic level. In both 12-week and term placentas hCG and hPL are present throughout the rough endoplasmic reticulum cisternae, in the Golgi bodies, and in the infrequent small dense granules of the syncytiotrophoblast. Previous assays have shown that hCG is at a higher concentration in early pregnancy and hPL peaks in late pregnancy, and our results corroborate these findings. No significant localisation of either hormone was seen in the cytotrophoblast or villous stroma. The results suggest that both hCG and hPL are synthesised and packaged by the classical secretory pathway, although the level of hormone stored in granules at any one time is small.
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  • 26
    ISSN: 1432-0878
    Keywords: GRF ; Immunocytochemistry ; Brain ; Pituitary ; Hypothalamus ; Vasotocin ; Anguilla anguilla ; Carassus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum to growth hormone-releasing factor (GRF) 1-44 was applied on brain and pituitary sections of nine teleost species. Immunoreactive (ir) perikarya were demonstrated in parvo- and magnocellular portions of the preoptic nucleus (PON) and occasionally in the nucleus lateralis tuberis. The two tracts originating in the PON ran ventro-laterally toward the optic chiasm and then caudally in the basal hypothalamus. In the pars distalis (PD) of the eel, carp, goldfish and salmonids, GRF-ir fibers did not enter the rostral PD and few fibers passed close to somatotropes. In.Myoxocephalus andMugil, a variable number of ir-fibers passed close to cells of the rostral and proximal PD. In the neurointermediate lobe, GRF-ir fibers were located exclusively in the neural tissue of the eel and trout. In goldfish, carp andMyoxocephalus, GRF-ir fibers entered the intermediate lobe. This antiserum also labeled corticotrops and, to a lesser extent, melanotrops in the pituitary of cyprinids. A variable number of perikarya contained both GRF and vasotocin in the PON of the eel. In all teleost species studied so far, the distribution patterns of GRF are different, and the function of the various adenohypophysial cell types appears to be differently modulated, according to the variable distribution of GRF in the pituitary.
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  • 27
    ISSN: 1432-0878
    Keywords: Bombesin-like peptide ; Immunocytochemistry ; Hypothalamus ; Neurosecretory ; Fibers ; Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and distribution of bombesin-like material were investigated in the brain of the cartilaginous fishScyliorhinus canicula using conventional immunocytochemical techniques. Perikarya containing bombesin-like immunoreactivity were identified in the hypothalamus, within the magnocellular component of the preoptic nucleus. Some immunopositive elements appeared to be of cerebrospinal fluid-contacting type. Beaded immunoreactive fibers were seen crossing the ventral telencephalon and the whole hypothalamus. An important tract of fibers was found in the infundibular floor and in the median eminence, in close contact with the vascular system of the pituitary portal plexus. A moderate number of positive fibers innervated the habenular complex and the dorsal wall of the posterior tuberculum. These findings indicate that a neuropeptide strictly related to amphibian bombesin is located in specific hypothalamic neurons ofS. canicula. The distribution of the immunoreactive fibers and terminals suggests that, in fish, this peptide, may be involved in neuroendocrine and neuromodulator functions.
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  • 28
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Development, ontogenetic ; Serotonin-like immunoreactivity ; identified neurons ; cerebral ganglion ; Tenebrio molitor (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serotonin-immunoreactive neurons in the brain of Tenebrio molitor L. have been demonstrated and mapped throughout metamorphosis. Most serotonin-immunoreactive brain neurons persist throughout metamorphosis; their fate can be followed during development because of their characteristic cell body locations and arborization patterns. The detailed morphology of the persisting neurons, however, changes during metamorphosis, probably to accommodate architectural changes of the different brain centers. Serotonin-immunoreactivity in the optic lobes allows a subset of neurons that is newly differentiated during metamorphosis to be identified. Phylogenetic homology of serotonin-immunoreactive brain interneurons of different insect species is discussed. The serotonin-immunoreactive brain neurons comprise a phylogenetically conserved neuronal population. Serial homologous abdomino-thoracic and brain serotonin-immunoreactive neurons were characterized, allowing a comparison of some basic structural features of these neurons.
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  • 29
    ISSN: 1432-0878
    Keywords: Cerebrospinal fluid-contacting neurons ; Paraventricular organ ; Posterior recess organ ; Somatostatin ; Serotonin ; Ultrastructure ; Immunocytochemistry ; Dogfish,Squalus acanthias skate,Raja radiata (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The paraventricular organ (PVO) and the posterior recess organ (PRO) of two elasmobranch species, the spiny dogfish,Squalus acanthias, and the skate,Raja radiata, were investigated by use of scanning and transmission electron microscopy and immunocytochemistry employing a series of primary antisera. The PVO and PRO contained four types of cerebrospinal fluid (CSF)-contacting neurons. One type was free of secretory granules and projected a dendrite-like process into the ventricle. The other three types were distinguished according to the size of their secretory granules. The ventricular extensions of these cells were filled with secretory granules. By means of immunocytochemistry three types of CSF-contacting neurons were observed in the PVO and PRO. Type I contained only serotonin; type 2 displayed only somatostatin; type 3 was endowed with both serotonin and somatostatin. Type I dominated in the PRO, whereas type 3 was the most frequent in the PVO. The latter cells appear to be the site of origin of a loose tract formed by serotonin- and somatostatinimmunoreactive fibers projecting from the PVO into the neuropil of the PRO. Compact bundles formed exclusively by serotonin fibers were also shown to extend between the PVO and PRO. The basal processes of the CSF-contacting neurons of the PRO penetrated into the underlying neuropil. This neuropil is rich in synapses and can be regarded as an integrative area to which the basal processes of the local CSF-contacting neurons, serotonin and somatostatin fibers from the PVO, and fibers containing immunoreactive thyrotropin-releasing hormone of unknown origin, support a conspicuous input. The present findings indicate that the PVO and PRO of elasmobranchs are functionally integrated structures.
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  • 30
    ISSN: 1432-0878
    Keywords: Oxytocin ; Corpus luteum ; Luteinized cyst ; Immunocytochemistry ; Ewe (Romney Marsh)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Anoestrous Romney Marsh ewes with or without progesterone pretreatment were injected with multiple low-doses of gonadotrophin-releasing hormone followed by a single, larger bolus. Blood samples were taken at twelve-hourly intervals for progesterone radioimmunoassay. Ewes were slaughtered on day 3 or 5 after the bolus injection, and the ovaries were collected for histology and immunocytochemical examination for oxytocin-immunocreactivity. The corpora lutea of all ewes killed on day 3 had similar weights and morphology. The ovaries of those ewes which were not pretreated with progesterone also contained some luteinized cysts. Ewes slaughtered on day 5 were separated into 2 groups according to plasma progesterone profiles, which were either rising (‘normal’), or falling after a transitory rise (‘abnormal’). Those ewes pretreated with progesterone all had a ‘normal’ progesterone profile whereas, of 14 ewes not pretreated with progesterone, 6 were ‘normal’ and 8 ‘abnormal’. Corpora lutea were significantly lighter in the ‘abnormal’ group and the ovaries of most of these ewes also contained luteinized cysts. All corpora lutea and luteinised cysts showed staining for oxytocin-immunoreactivity although the staining intensity was variable. In corpora lutea from ‘normal’ ewes oxytocin was restricted to large luteal cells. In addition tissues from ‘abnormal’ ewes also contained many cells with an atypical elongated shape which stained for oxytocin-immunoreactivity. These results show that progesterone pretreatment is needed for both normal morphological and endocrine development of corpora lutea in anoestrous ewes stimulated with gonadotrophin-releasing hormone.
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  • 31
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    Cell & tissue research 262 (1990), S. 569-577 
    ISSN: 1432-0878
    Keywords: Blastocyst ; Uterine proteins ; Uteroglobin ; Endocytosis ; Autoradiography ; Immunocytochemistry ; Acid phosphatase ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Uptake of uteroglobin (UGL) by day-6 rabbit blastocysts and the intracellular fate of this protein were studied by light- and electron-microscopic autoradiography, immunocytochemistry and acid-phosphatase cytochemistry. UGL, labelled with N-succinimidyl-(2-3-3H)-propionate, was administered to embryos in vitro for 15 min to 4 h. The kinetics, determined from light-microscopic autoradiographs, showed a continuous uptake of the labeled protein over a 4-h period of incubation. At the ultrastructural level, increasing numbers of silver grains and an intense UGL immunoreaction in protein vacuoles and crystalloid bodies of trophoblast cells indicated that 3H-UGL had accumulated in these organelles. The presence of crystalloid inclusions in protein vacuoles suggests their origin by a condensation of the protein content, including UGL. Lysosomes containing radioactivity were rarely found, suggesting a very low degradation rate of the 3H-UGL. Protein vacuoles and crystalloid bodies exhibited no acid phosphatase reaction. The enzyme was mainly found outside the basal and lateral cell membranes of trophoblast cells, and on the rough endoplasmic reticulum of endoderm cells.
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  • 32
    ISSN: 1432-0878
    Keywords: Median eminence ; Catecholamines ; Neuropeptides ; Immunocytochemistry ; Double labeling ; Ultrastructure ; Triturus alpestris (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Dopaminergic and peptidergic nerve fibers were simultaneously demonstrated with a double-labeling technique at the ultrastructural level. The first antibody, raised against tyrosine hydroxylase, was applied during the preembedding phase and visualized with the peroxidase method. The second antibody, raised against one of the peptides met-enkephalin, somatostatin or gonadotropin-releasing hormone (GnRH), was applied to the ultrathin sections and visualized with gold-labeled goat anti-rabbit IgG. The fibers of both categories were present in the zona externa of the median eminence, frequently contacting the basal lamina of the portal vessels. In addition, topographical relationships between different types of nerve fibers were observed in the perivascular areas, although there were no morphological signs of synaptic specializations. Using serial sections, it could be established that one GnRH-fiber contacted both a dopaminergic fiber and a fiber immunoreactive for met-enkephalin. The observations support earlier physiological data concerning the regulation of the hypothalamo-hypophyseal axis, with special emphasis on the release of neurohormones in the median eminence of the newt.
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  • 33
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    Cell & tissue research 260 (1990), S. 381-386 
    ISSN: 1432-0878
    Keywords: Molluscan insulin-related peptide ; Neuropeptide ; Immunocytochemistry ; In situ hybridization ; Lymnaea stagnalis (Mollusca) ; Aplysia californica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary the occurrence of insulin-related substances in the central nervous system of pulmonates and Aplysia californica was investigated by means of immunocytochemistry and in situ hybridization. Previous experiments have shown that, in Lymnaea stagnalis, the growth hormone-producing neurons in the cerebral ganglia (the so-called light green cells) express at least 5 genes that are related to the vertebrate insulin genes, i.e., they encode prohormones that are composed of a B- and A-chain and a connecting C peptide. These insulin related molecules also have the amino acids essential for their tertiary structure (viz. cysteines) at identical positions to those of the vertebrate insulins. In the investigated basommatophoran and stylommatophoran snails and slugs, neurons reacted with an antiserum raised against the C peptide of one of the molluscan insulin-related peptides. These neurons can be considered to be, based on morphological and endocrinological criteria, homologous to the light green cells of L. stagnalis. In A. californica, all central ganglia contain immunoreactive neurons. The highest number (about 50) was observed in the abdominal ganglion. The present results indicate that insulin-related substances are generally occurring neuropeptides in the central nervous system of molluscs.
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  • 34
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    Cell & tissue research 262 (1990), S. 105-113 
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Lectin histochemistry ; Immunocytochemistry ; Glycoproteins ; Human fetuses ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO) of 7 human fetuses, 3 to 6.5 months old, was investigated by means of: (i) immunocytochemistry employing three different antisera against secretory products extracted from the bovine SCO and Reissner's fiber; (ii) lectin binding using concanavalin A (Con A; affinity: mannose, glucose), wheat-germ agglutinin (WGA; affinity: N-acetyl-glucosamine, sialic acid), and Limax flavus agglutinin (LFA; affinity: sialic acid). Sections of bovine SCO were processed simultaneously and examined for comparative purposes. The human fetal SCO displayed lectin-binding properties identical to those in the SCO of other mammals. Thus, Con-A-binding sites were restricted to abundant supranuclear structures that most likely corresponded to the rough endoplasmic reticulum, but were missing from granules located in the apical cytoplasm. The latter secretory material was strongly WGA- and LFA-positive and formed a distinct zone in the most apical portion of the ependymal cells. In contrast, this type of reactivity was missing in the adjacent cells of ependyma proper. In the bovine SCO, LFA-positive granules were also aggregated in an apical layer. The secretory material in the bovine SCO, especially its apical granular component, was strongly immunoreactive with the three antisera used; the human fetal SCO, however, lacked this immunoreactivity. It is postulated that the SCO of human fetuses secretes glycoproteins with a carbohydrate chain similar to-and a protein backbone different from-the secretions elaborated by the SCO of other vertebrate species.
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  • 35
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    Cell & tissue research 260 (1990), S. 431-434 
    ISSN: 1432-0878
    Keywords: Actin ; Cytoskeleton ; Immunocytochemistry ; Photoreceptor cells ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The Drosophila ninaC mutation produces small rhabdomeres with the axial filament of the microvillar cytoskeleton reduced or missing. Using post-embedding immunogold labelling of LR White-embedded eyes, we show that several alleles of this mutation retain positive anti-actin immunoreactivity in the rhabdomeres, comparable to that of wild-type flies.
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  • 36
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    Cell & tissue research 261 (1990), S. 461-466 
    ISSN: 1432-0878
    Keywords: Secretion ; Plasmalemma ; Membrane dynamics ; Immunocytochemistry ; Freeze-fracturing ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The movements of the molecular components of the luminal plasma membrane during exocytotic secretion in parotid acinar cells were examined. For immunocytochemical study, we used an antiserum of dipeptidyl peptidase IV as a marker for the components of the luminal plasma membrane of acinar cells. In unstimulated acinar cells, dipeptidyl peptidase IV immunoreactivity is restricted to the luminal plasma membrane. However, after secretion was stimulated with a β-adrenergic agonist, isoproterenol, immunostaining became detectable on the membrane of discharged granules. Freeze-fracture images showed that the density of intramembrane particles on the P-fracture leaflets of discharged granule membranes is much higher than that of undischarged granule membranes during secretion. These results suggest that in parotid acinar cells of the rat, the components of the luminal plasma membrane move laterally, during secretion, to the membranes of discharged granules.
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  • 37
    ISSN: 1432-0878
    Keywords: Serotonin (5-HT) ; Substance P ; GABA ; Immunocytochemistry ; Invertebrate peripheral nervous system ; Mytilus galloprovincialis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical methods were applied to study the distribution of putative neurotransmitters (5-HT, substance P, GABA, glutamate and aspartate) in the nerve plexuses of the foot and the anterior byssus retractor muscle (ABRM) of Mytilus galloprovincialis (Mollusca, Bivalvia). The foot presents extensive nerve plexuses containing 5-HT and substance P-like immunoreactive material with a similar distribution beneath the surface epithelium, around the vessels and in the glandular regions. Coexistence of the two putative neurotransmitters was observed in a few nerve fibers, Conversely, muscle fibers, both in the foot and in the ABRM, are innervated only by 5-HT-positive fibers, while substance P-like material is present only in the networks of the ABRM epimysial sheath. Immunoreactivity for glutamate and aspartate was not demonstrated, while rare GABA-positive nerve cells and fibers were found only in the foot. The results of this investigation provide a morphological background to previous physiological studies on 5-HT in the nervous system of bivalve molluscs. Moreover, they confirm that the nervous system of Mytilus contains a remarkable amount of a substance related to the vertebrate tachykinin family.
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  • 38
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    Cell & tissue research 261 (1990), S. 517-524 
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Adrenal medulla ; Somatostatin immunoreactivity ; Immunocytochemistry ; Hybridization, in situ ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous studies have shown that somatostatin modulates angiotensin-induced aldosterone secretion by adrenal glomerulosa cells. This effect is mediated through specific receptors which do not show any preference for somatostatin-14 (S14) or the N-extended form somatostatin-28 (S28). The study of the distribution of 125I-Tyr [Tyr0, DTrp8] S14-and 125I-Tyr[Leu8, DTrp22, Tyr25] S28-binding in frozen sections of the rat adrenal by autoradiography indicated that both peptides bind to similar loci. High concentrations of binding sites were observed in the zona glomerulosa, and low concentrations were detected in the medulla. At the ultrastructural level, immunocytochemistry after cryoultramicrotomy revealed endogenous S14-and S28-like immunoreactive material in zona glomerulosa and in medulla. In glomerulosa cells, immunoreactive material was localized at the plasma membrane level, in the cytoplasmic matrix, in the mitochondria, and in the nucleus. S14-and S28-like materials were detected in both epinephrine and norepinephrine-storing cells of the adrenal medulla. In these cells, the distribution of either immunoreactive product was similar; it was observed in cytoplasmic matrix, secretory granules and nucleus, but not at the plasma membrane level. In situ hybridization does not reveal somatostatin mRNA in zona glomerulosa or medulla. These results demonstrate that S14 and S28 bind to, and are taken up by zona glomerulosa and adrenal medullary cells, but are not produced by these cells.
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  • 39
    ISSN: 1432-0878
    Keywords: Neuropeptides (pancreatic polypeptide, peptide YY, neuropeptide Y) ; Immunocytochemistry ; Confocal scanning laser microscopy ; Schistosoma mansoni (Scolecida, Trematoda)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and distribution of neuropeptides belonging to the pancreatic polypeptide family have been demonstrated by an indirect immunofluorescence technique in the nervous systems of adult male and female Schistosoma mansoni. Seven antisera of differing regional specificity to pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY) were employed on both whole-mount and cryostat-sectioned material. Positive immunoreactivity (IR) was obtained with all antisera except an N-terminally-directed antiserum to NPY. In the CNS, immunoreactivity was restricted to cell bodies and nerve fibres in the anterior ganglia, central commissure and dorsal and ventral nerve cords of both sexes, whereas, in the PNS, positive-IR was present in the plexuses innervating the subtegumental musculature and the oral and ventral suckers. Intense immunoreactivity was observed in a plexus of nerve fibres and cell bodies in the lining of the gynaecophoric canal and in fine nerve fibres innervating the dorsal tubercles of the male. In contrast, in the female, strong immunoreactivity was evident in nerve plexuses innervating the lining of the ovovitelline duct and in the wall of the ootype, but most notably in a cluster of cells in the region of Mehlis' gland. Results suggest that molecules with C-terminal homology to the PP-family are present in S. mansoni. These peptides would appear to be important regulatory molecules in the parasite's nervous system and may play a role in the control of egg production.
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  • 40
    Electronic Resource
    Electronic Resource
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    Cell & tissue research 262 (1990), S. 115-123 
    ISSN: 1432-0878
    Keywords: ACTH ; Immunocytochemistry ; Brain ; Hypothalamus ; Corticotropin-releasing factor ; Pituitary ; Carassius auratus, Salmo fario, Anguilla anguilla (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunostaining of brain and pituitary sections of three teleost species (goldfish, trout and eel) with antisera to porcine and human ACTH 1–39 revealed the presence of an ACTH (adrenocorticotropic hormone)-like peptide in the ventral hypothalamus. Perikarya were localized in the rostral, median and posterior portions of the nucleus lateralis tuberis (NLT); some were in contact with the cerebrospinal fluid. A dense network of immunoreactive (ir) fibers occurred in the peri-infundibular region and extended into the periventricular tissue, around the lateral and posterior recesses. Rostrally directed ir-fibers reached the telencephalon either ventrally or mediodorsally; some were observed in the olfactory lobe. In the mesencephalon, ir-fibers penetrated into the optic tectum of the goldfish. In the pituitary, both antisera intensely labeled rostral ACTH cells. Small groups of labeled cells were scattered in the rostral pars distalis and the proximal pars distalis. A gradient of activity was evident among ACTH cells: those located along the rostral neurohypophysis containing corticotropin-releasing factor nerve terminals were larger and often more marked than those farther away from the neural tissue. ACTH-like peptide in the brain may act as a neuromodulator, mainly in the NLT and the preoptic nucleus, and around the nuclei of the ventricular recesses containing serotonin and catecholamines.
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  • 41
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    Electronic Resource
    Springer
    Protoplasma 158 (1990), S. 109-120 
    ISSN: 1615-6102
    Keywords: Differentiation ; Immunocytochemistry ; [3H]T-autoradiography ; Stem cells ; Turbellaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A combination of microscopical, immunocytochemical, and autoradiographic techniques were employed to study stem cells and their fates during asexual reproduction and regeneration in two microturbellarians,Microstomum lineare (Macrostomida) andStenostomum leucops (Catenulida). Special attention was paid to the development of the immunoreactivity (IR) to FMRF/RF-amide and 5-HT in differentiating nerve cells. Asexual reproduction inM. lineare andS. leucops occurs by paratomy, i.e., fragmentation after completed differentiation of the new organs. Regeneration, on the other hand, involves a combination of morphallactic and epimorphic processes without the formation of a regeneration blastema. The only cells incorporating tritiated thymidine ([3H]T) were the mesenchymal and gastrodermal neoblasts, which proliferate continuously replenishing the population of stem cells available for growth, asexual reproduction and regeneration. These proliferative cells occurred in two ultrastructurally different forms, differing from each other only by the presence or absence of ciliar basal bodies in the cytoplasm. Few differentiated cells were labeled in the head piece after completed regeneration. A greater amount of labeled differentiated cells were, however, observed postpharyngeally in the first zooid as well as in zooids having developed during the same time (i.e., 20–45 h after the treatment with [3H]T). Furthermore, many labeled cells were still undifferentiated at that time or just in the beginning of the differentiation process. It can therefore be concluded that neoblasts function both as reserve cells and as functional stem cells for all differentiated cell types in these worms. IR to FMRF/RF-amide neuropeptides was not observed in nerve cells differentiating from neoblasts until the occurrence of dense-core vesicles in their cytoplasm. Due to methodological difficulties only weak or no IR to 5-HT could be traced in the nervous system of the asexual and regenerating worms.
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  • 42
    ISSN: 1615-6102
    Keywords: α-Amylase isozymes ; Barley aleurone ; Endoplasmic reticulum ; Golgi apparatus ; Immunocytochemistry ; Intracellular transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The localization of α-amylase (EC 3.2.1.1) in barley (Hordeum vulgare L. cv Himalaya) aleurone protoplasts was studied using electron microscope immunocytochemistry. Antibodies were raised against total barley α-amylase, i.e., α-amylase containing both highisoelectric point (high-pI) and low-pI isoforms, as well as against purified high- and low-pI isoforms. All antibodies localized α-amylase to the endoplasmic reticulum (ER) and Golgi apparatus (GApp) of the aleurone cell, and various controls showed that the labeling was specific for α-amylase. Labeling of protein bodies and spherosomes, which are the most abundant organelles in this cell, was very low. There was no evidence that α-amylase isoforms were differentially distributed within different compartments of the endomembrane system. Rather, both high- and low-pI isoforms showed the same pattern of distribution in ER and in the cis, medial, and transregions of the GApp. We conclude that in the Himalaya cultivar of barley, all isoforms of α-amylase are transported to the plasma membrane via the GApp.
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  • 43
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 160 (1990), S. 119-126 
    ISSN: 1432-136X
    Keywords: Atrial natriuretic peptide ; Immunocytochemistry ; Radioimmunoassay ; Hypotension ; Anguilla japonica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cardiocytes positive for human atrial natriurectic peptide (hANP) were identified histochemically in the eel atrium, but they were not found in the ventricule. Secretory granules were frequently observed in atrial cardiocytes by electron microscopy, but the number of such granules was quite small in the ventricle. Immunogold cytochemistry revealed that immunoreactive ANP (IR-ANP) in atrial cardiocytes was localized in these granules. In spite of poor immunostaining of the eel ventricle, an acid extract of the ventricle contained 25±4 ng·g tissue-1 (n=9) of IR-ANP when the level of IR-ANP was measured by radioimmunoassay (RIA) for hANP. This level was one eight of that measured in atrial extracts (203±13 ng·g tissue-1, n=9). Plasma contained 116.7±18.6 pg·ml-1 (n=9) of IR-ANP. An extract of eel hearts decreased arterial pressure in eels and quail as did hANP. The level of ANP in the extract, as measured by an eel vasodepressor bioassay, was much greater than that measured by RIA for hANP. The immunoreactive and bioactive ANP in the heart extract are identical since the vasodepressor activity disappeared after IR-ANP was absorbed by excess antibodies raised against hANP. Chromatography on Sephadex G-75 generated a major peak of IR-ANP at a position that corresponded to a molecular weight of 14 kD and minor peaks at 3–7 kD from both plasma and heart extract. Reverse phase HPLC of plasma and heart extract generated several peaks of IR-ANP at positions more hydrophilic than those of mammalian ANPs. These results show that eel hearts contain immunoreactive and bioactive ANPs which are distinctly different from hANP. These ANPs are synthesized both in the atrium and in the ventricle, and they are secreted into the circulation mostly in the larger molecular form. The atrial ANP may be stored in the granules and secreted upon exposure of eels to certain stimuli, but the ventricular ANP may be secreted constitutively into the circulation without prior storage in the granules.
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