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  • Immunocytochemistry  (149)
  • Springer  (149)
  • Elsevier
  • Oxford University Press
  • 1980-1984  (149)
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Year
  • 1
    ISSN: 1432-041X
    Keywords: Major haemolymph proteins ; Development ; Cuticle ; Immunocytochemistry ; Ceratitis capitata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The developmental profile of the major haemolymph proteins (ceratitins) inCeratitis capitata was studied. Ceratitin concentration in the haemolymph decreases dramatically during the last days of pupal life, while the amounts of ceratitins in whole organism extracts remain unchanged. By electrophoretic, immunological and immunofluorescence techniques it was revealed that ceratitins are reabsorbed by the fat body and a fraction of them is deposited in the cuticle. The possible role of ceratitins is discussed.
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  • 2
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; Lectin (localization) ; Phaseolus (lectin) ; Phytohemagglutinin ; Seed (lectin)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have examined the properties and subcellular localization of phytohemagglutinin (PHA), the major lectin of the common bean (Phaseolus vulgaris.), in the axis cells of nearly mature and imbibed mature seeds. On a protein basis the axis contained about 15% as much PHA as the cotyledons. Localization of PHA was done with an indirect immunolabeling method (rabbit antibodies against PHA, followed by colloidal gold particles coated with goat antibodies against rabbit immunoglobulins) on ultra-thin cryosections which were embedded in plastic on the grids after the immunolabeling procedure. The embedding greatly improved the visualization of the subcellular structures. The small (4 nm) collodial gold particles, localized with the electron microscope, were found exclusively over small vacuoles or protein bodies in all the cell types examined (cortical parenchyma cells, vascular-bundle cells, epidermal cells). The matrix of these vacuoles-protein bodies appears considerably less dense than that of the protein bodies in the cotyledons, but the results confirm that in all parts of the embryo PHA is localized in similar structures.
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  • 3
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    Protoplasma 121 (1984), S. 25-33 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Colloidal gold ; Wheat germ agglutinin ; Lectin ; Cryosections
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chitin-binding lectin wheat germ agglutinin (WGA) is found at the periphery of wheat embryos, and a similar lectin is present at the root tips of older plants (Mishkind et al. 1982). Although a ferritin-conjugated secondary antibody is adequate for localizing WGA in embryos, native electron-opaque particles make the electron microscope identification of added label equivocal in other wheat tissues. As reported here, however, unambiguous ultrastructural localization of WGA-like lectin in adult wheat roots can be obtained with rabbit anti-WGA followed by colloidal gold-labeled goat anti-rabbit (GAR) IgG. Colloidal gold (CG) was prepared by the reduction of gold chloride with citrate, ascorbate or phosphorous. GAR IgG, prepared from serum by antigen affinity chromatograhy, was adsorbed to the gold particles to produce a stabilized suspension of GAR-CG. Localization was performed on 8–12 μM frozen sections of tissue fixed in 4% paraformaldehyde, 0.3% glutaraldehyde, and 0.75% acrolein in phosphate-buffered saline containing 1M sucrose. Localization with GAR-CG was first compared to that ascertained in embryos using other probes and was then extended to the roots of adult plants. An advantage of the GARCG method is that it permits the visualization of antigen at both the light and electron microscope levels in the same section. At the light level, the anti-WGA-GAR-CG complex appears as a red stain that is localized in specific tissues of embryos and in the caps and outer layers of adult roots. Sections in which lectin was detected at the light microscope level were embedded in plastic and sectioned for subcellular examination. Electron dense gold particles indicative of WGA are found at the periphery of protein bodies in wheat embryos and in vacuoles of the roots of adult plants. Sections incubated with control IgG lack reaction product.
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  • 4
    ISSN: 1615-6102
    Keywords: Bauhinia purpurea ; Colloidial gold ; Golgi apparatus ; Immunocytochemistry ; Lectin ; Lowicryl K4M ; Protein body
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The seed lectin of the tree legume,Bauhinia purpurea alba, was localized by electron microscopic immunocytochemistry. The pattern of lectin deposition and site of intracellular localization was examined in mid- to late-maturation seeds. The seed tissue was embedded in Lowicryl K4M, the use of which with seed tissues is discussed. Immunocytochemical labeling was accomplished with colloidal gold coupled to a second antibody. The immunocytochemical reaction was specific and sensitive. Protein bodies, Golgi apparatus and Golgi secretion vesicles were densely labeled. Golgi apparatus was oriented such that Golgi secretion vesicles were in close proximity to the protein bodies. The entire Golgi apparatus was labeled with no concentration gradient across the Golgi stack. These observations indicate that the final site of lectin deposition is the protein body, and that the Golgi apparatus plays an essential role in the deposition process.
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  • 5
    ISSN: 1432-0878
    Keywords: Urotensin ; Caudal neurosecretory system ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of urotensin I has been investigated in the caudal neurosecretory system of the white sucker (Catostomus commersoni). The peptide is present in all the cells of the system both large and small, in the large axons passing to the urophysis, and in fine beaded fibres not only within the urophysis but also in a fine plexus lateral to the large cells in the spinal cord proper. The possibility that the caudal neurosecretory system is not a functionally uniform system but rather a collection of dissimilar cells of different synaptic inputs with a common entity, urotensin I, is discussed. Moreover, the feasibility of a urotensin I feedback loop is described.
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  • 6
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    Cell & tissue research 235 (1984), S. 211-214 
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Pars distalis ; Immunocytochemistry ; Amphibia ; Gymnophiona
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunofluorescence method was used to identify and locate LTH-, STH-, LH-, TSH-, ACTH- and MSH-immunoreactive cells in the pituitary of Typhlonectes compressicaudus (Gymnophiona). The present study defines the histological and histochemical staining properties of each cell type identified.
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  • 7
    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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  • 8
    ISSN: 1432-0878
    Keywords: FMRFamide ; Bovine pancreatic polypeptide ; Immunocytochemistry ; Peptidergic neurons ; Leptinotarsa decemlineata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Particular neurons in the nervous system of the Colorado potato beetle, Leptinotarsa decemlineata, are recognized by antisera against bovine pancreatic polypeptide and FMRFamide. Both antisera react with the same neurons. Solid phase absorptions showed that antiserum against bovine pancreatic polypeptide cross-reacts with FMRFamide, whereas antiserum against FMRFamide cross-reacts with bovine pancreatic polypeptide. Some of the immunoreactive neurons have axons branching extensively within the neuropile, which suggests that the peptide is used as transmitter. In the corpus cardiacum, a neurohaemal organ in insects, numerous immunoreactive axon terminals are present. Here, the peptide material is presumably released as a hormone.
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  • 9
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    Cell & tissue research 235 (1984), S. 427-432 
    ISSN: 1432-0878
    Keywords: DPP IV ; Salivary glands ; Ultrastructural localization ; Immunocytochemistry ; PAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of dipeptidyl peptidase IV (DPP IV) (EC 3.4.14.5) in rat submandibular and parotid glands was studied immunocytochemically by the peroxidase-antiperoxidase (PAP) method, using a monospecific antiserum against rat kidney DPP IV. There were no differences in the immunocytochemical localization of DPP IV between submandibular and parotid glands. In these glands, DPP IV was primarily found to be associated with the luminal and intercellular canalicular plasma membranes of acinar cells and with the luminal plasma membranes of intercalated and striated duct cells. Occasionally, immunoreaction of DPP IV was detected in cytoplasmic vesicles (vacuoles), lysosomes, and multivesicular bodies in some acinar cells as well as in ductal epithelial cells. Furthermore, the reaction product was also found within the lumina of peri-acinar and peri-ductal capillaries and in the cytoplasm of some fibroblasts in the interstitial connective tissue. These data suggest that DPP IV in the submandibular and parotid glands may play some role in the secretion or reabsorption processes of secretory proteins and peptides in these glands.
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  • 10
    ISSN: 1432-0878
    Keywords: S-100 protein ; Thymus ; Interdigitating cells ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study deals with the localization and ultrastructure of S-100-immunoreactive cells in the human thymus. These immunoreactive cells are distributed mainly in the medulla with some scattered elements in the cortex. Electron-microscopic observation revealed that the cells are characterized by an irregularly shaped nucleus, tubulovesicular structures in the cytoplasm and characteristic interdigitations of the plasma membrane. The cells often embrace lymphocytes with their branched processes. On the basis of these morphological features, the immunostained elements were identified as interdigitating cells (IDCs). The immunocytochemistry for S-100 visualizes the precise distribution and extension of the IDCs under the light microscope and indicates that the IDCs form no structural networks such as those established by the thymic epithelial cells. Since the IDCs in human lymph nodes have also been reported to contain S-100-like immunoreactivity, S-100 protein can be regarded as a useful marker for identifying the IDCs in the human thymus and other lymphoid organs.
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  • 11
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    Protoplasma 122 (1984), S. 86-90 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Non-articulated laticifers ; Articulated laticifers ; Asclepiadaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary sensitive immunocytochemical method for the identification of laticifers has been developed. Frozen sections of various laticifer-bearing plant material, mounted on slides, were first flooded with the IgG fraction of rabbit anti-latex antiserum, prepared using whole latex ofAsclepias syriaca, then flooded with fluorescein-conjugated IgG fraction goat anti-rabbit IgG to visualize laticifers. Positive fluorescence was observed for laticifers in shoots and embryos ofA. syriaca andStapelia bella and embryos ofA. tuberosa. Laticifers did not fluoresce in shoots ofA. tuberosa andEuphorbia tirucalli, in embryos ofE. marginata, or in petioles ofMusa paradisiaca andCichorium intybus. Controls prepared with uninjected rabbit serum were negative (no fluorescence).
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  • 12
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; FMRF-amide ; Neurotransmitter ; Palaemon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of a specific antiserum against the molluscan cardio-excitatory tetrapeptide FMRF-amide in combination with the PAP-method it was possible to obtain positive immunocytochemical reactions in several neurosecretory regions of the eyestalk of the prawn Palaemon serratus. FMRF-amide-like material was found in perikarya and nerve fibers of the medulla terminalis and in neurons in the lamina ganglionaris. The immunoreactivity observed in the glandular tissue located at the basal insertion of the eyestalk muscles must be ascribed to a non-specific reaction. The identification of immunopositive nerve fibers, ending on a nerve bundle in the medulla terminalis, and the fact that immunoreactive material was absent in the neurohemal sinus gland seem to indicate a neurotransmitter/neuromodulator function.
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  • 13
    ISSN: 1432-0878
    Keywords: Fetal porcine pituitary ; ACTH, MSH, β-LPH, α- and β-endorphin, GH, PRL ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The aim of the present study on the fetal porcine pituitary was (1) to detect by means of the immunoperoxidase technique the earliest stages of cells producing polypeptide hormones: β-MSH, ACTH, β-LPH, α- and β-endorphin, growth hormone (GH) and prolactin (PRL), (2) to study the development of the synthesis and the storage of these hormones during fetal life, and (3) to detect whether several hormones can be located in one and the same cell. The corticotropic cells were revealed as the earliest functional elements of the fetal anterior pituitary. Our results indicate clearly that ACTH, β-MSH, β-LPH, α- and β-endorphin appear at 34 days in the same regular, round or ovoid cells; no differences in the time of their appearance could be observed. The ACTH-cells, irregular or angular in shape and endowed with cytoplasmic processes such as described in the adult pituitary, were not seen until day 50. The first GH-cells were detected between 40 to 45 days of fetal life. From day 45 to 90, the GH-cells greatly increased in number and in staining intensity of their progressively extending cytoplasmic area, but they displayed the same regular and round shape. The PRL-cells were the last cell type to appear in the fetal pituitary. The first PRL-cells, small in size and round or ovoid in shape with a high nucleus/cytoplasm ratio, were detected at day 70. At day 80, the PRL-cells increased in size and staining intensity. They displayed an irregular elongated or stellated shape and cytoplasmic processes resembling those characteristic of the adult pituitary. These data suggest that in the fetal porcine pituitary: (1) ACTH, β-LPH and related peptides are synthesized and stored in the same cells, and (2) PRL and GH appear in individual cellular elements.
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  • 14
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    Cell & tissue research 236 (1984), S. 517-525 
    ISSN: 1432-0878
    Keywords: Bone marrow ; Actin ; Cell motility ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blood cells proliferate extravascularly in the bone marrow and enter the circulation by migrating through endothelial cells of venous blood sinuses. This migration, or diapedesis, was suspected to involve actin. To test for the presence and distribution of actin, sections of rat bone marrow were examined by indirect immunocytochemistry. Affinity purified rabbit antichicken gizzard actin antibody, and goat-antirabbit IgG-FITC, or goat antirabbit IgG colloidal gold probes were used. The migrating cell contacts the endothelial cell and forms a podosome (a cortical bleb). Immunocytochemistry shows this region to contain actin. As diapedesis proceeds the podosome deforms, then breaches the endothelial cell. At this time the anterior portion of the leukocyte shows heavy labeling for actin. When the migratory cell traverses approximately half of its length through the endothelial cell, actin appears prominent in the caudal region of the cell. The immunocytochemical data suggest that actin is nonrandomly distributed in leukocytes undergoing diapedesis and may be a component of the force-generating mechanism responsible for this transcellular migratory event.
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  • 15
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Reissner's fiber ; Ependyma ; Secretory process ; Comparative analysis ; Immunocytochemistry ; Vertebrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organs (SCO) of 76 specimens belonging to 25 vertebrate species (amphibians, reptiles, birds, mammals) were studied by use of the immunoperoxidase procedure. The primary antiserum was obtained by immunizing rabbits with bovine Reissner's fiber (RF) extracted in a medium containing EDTA, DTT and urea. Antiserum against an aqueous extract of RF was also produced. The presence of immunoreactive material in cell processes and endings was regarded as an indication of a possible route of passage. Special attention was paid to the relative development of the ventricular, leptomeningeal and vascular pathways established by immunoreactive structures. The SCO of submammalian species is characterized by (i) a conspicuous leptomeningeal connection established by ependymal cells, (ii) scarce or missing hypendymal cells, and (iii) a population of ependymal cells establishing close spatial contacts with blood vessels. The SCO of most mammalian species displays the following features: (i) ependymal cells lacking immunoreactive long basal processes, (ii) hypendymal secretory cells occurring either in a scattered arrangement or forming clusters, (iii) an occasional leptomeningeal connection provided by hypendymal cells, and (iv) in certain species numerous contacts of secretory cells with blood vessels. In the hedgehog immunoreactive material was missing in the ependymal formation of the SCO, but present in hypendymal cells and in the choroid plexuses. The SCO of several species of New-and Old-World monkeys displayed immunoreactive material, whereas that of anthropoid apes (chimpanzee, orangutan) and man was completely negative with the antisera used.
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  • 16
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Ependyma ; Comparative aspects ; Immunocytochemistry ; Secretory process ; Blood vessels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In 76 specimens (amphibians, reptilians, mammals) belonging to 25 different vertebrate species, the region of the subcommissural organ (SCO) was investigated with the use of a primary antiserum raised against an extract of bovine Reissner's fiber+the immunoperoxidase procedure according to Sternberger et al. (1970). In the SCO of a toad (Bufo arenarum) and several species of reptiles (lacertilians, ophidians, crocodilians), the ependymal cells were the only type of secretory cell displaying vascular contacts, whereas in mammals ependymal and hypendymal cells established intimate spatial contacts with blood vessels. In Bufo arenarum, but especially in the reptilian species examined, the ependymo-vascular relationship was exerted by a population of ependymal cells having a rather constant location within the SCO and projecting to capillaries that showed a remarkably constant pattern of anatomical distribution. In the SCO of mammals the modality and degree of the structural relationships between secretory cells and blood vessels varied greatly from species to species. In the SCO of the armadillo and dog the secretory tissue was organized as a thick, highly vascularized layer with most of the cells oriented toward the capillaries. A rather opposite situation was found in the SCO of New-and Old-World monkeys, where vascular contacts were restricted to a few ependymal cells.
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  • 17
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    Cell & tissue research 237 (1984), S. 521-524 
    ISSN: 1432-0878
    Keywords: α-Melanocyte-stimulating hormone ; α-MSH-like peptide ; Immunocytochemistry ; Hypothalamus ; Lizard (Lacerta muralis)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of α-melanocyte-stimulating hormone (α-MSH) was studied in the brain of the lizard Lacerta muralis by means of immunocytochemical staining methods. α-MSH-like containing cells were found in the ventro-lateral preoptic area and the paraventricular and supraoptic nuclei. Some scattered cells staining for α-MSH were also detected in the mesencephalo-diencephalic boundary region, while numerous α-MSH-like nerve fibres were localized in the medial eminence. No reaction was observed after the use of antiserum preabsorbed with synthetic antigen. These findings suggest that an α-MSH-like peptidergic system could possibly be involved in the hypothalamo-hypophysial regulation and/or play a role as neurotransmitter in this animal.
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  • 18
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    Cell & tissue research 238 (1984), S. 203-205 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Hypothalamus, human ; Immunocytochemistry ; Pituitary stalk
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to study the distribution of neuropeptide Y-like immunoreactivity in the human hypothalamus, an immunocytochemical localization of this peptide was performed. Using antibodies developed against synthetic porcine neuropeptide Y (NPY), we have been able to localize immunoreactivity in neuronal cell bodies located exclusively in the infundibular nucleus. Immunostained fibers were found in several regions in the hypothalamus with a high concentration in the periventricular areas. Fibers were also found in the neurovascular zone of the median eminence, the pituitary stalk and the posterior pituitary. These results suggest that immunoreactive material related to porcine NPY is present in the human hypothalamus, with a distribution similar to that observed in the rat.
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  • 19
    ISSN: 1432-0878
    Keywords: Peptidergic neurotransmission ; Lymnaea stagnalis ; Immunocytochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three neuronal systems of the pond snail Lymnaea stagnalis were immunocytochemically investigated at the ultrastructural level with the unlabeled peroxidase-antiperoxidase technique. Preliminary electrophysiological and cell-filling investigations have shown that a cluster of neurons which reacts positively with an antiserum against the molluscan cardio-active peptide FMRFamide, sends axons to the penis retractor muscle. In this muscle anti-FMRF-amide (aFM) positive axons form neuro-muscular synapses with (smooth) muscle fibers. The morphological observations suggest the aFM immunoreactive system to be involved in peptidergic neurotransmission. In the right parietal ganglion a large neuron (LYAC) is penetrated by aFM positive axons which form synapse-like structures (SLS) with the LYAC. The assumption that the SLS represent the morphological basis for peptidergic transmission is sustained by the observation that iontophoretical application of synthetic FMRFamide depolarizes the LYAC. The axons of a group of pedal anti-vasopressin (aVP) positive cells run in close vicinity to the cerebral ovulation (neuro-)-hormone producing cell system (CDC system) Synapses or SLS between the two systems were not observed. The fact that (bath) application of arg-vasopressin induces bursting in the CDC, may indicate that the vasopressin-like substance of the aVP cells is released non-synaptically.
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  • 20
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    Cell & tissue research 236 (1984), S. 171-180 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuropil located ventral to the SON was investigated by the use of immunoperoxidase staining for neurophysins, oxytocin and vasopressin, and electron miroscopy. The study was performed in six groups of rats: 1) control; 2) infusion of isotonic saline into the CSF; 3) infusion of hypertonic saline into the CSF; 4) drinking hypertonic saline for 4 days; 5) same as group 4 but injection of colchicine into the CSF on second day of dehydration; 6) salt loading for 3 months. In the control rats the ventral neuropil contained a few immunoreactive processes, the general morphology of which was completely different from that of the neurosecretory axons emerging from the SON at its dorsal aspect. In rats of groups 3 to 6 the ventral processes (VP) became loaded with neurosecretory granules, whereas the perikarya and axons were depleted. Based on their general morphology and reactivity pattern it is suggested that the VP are dendrites. Most of these “dendrites” were embedded in a glial cushion formed by the processes of a particular type of marginal glia. Some of these “dendrites” enveloped an arteriole penetrating the optic tract. All VP were rich in synaptic contacts. The possibility that the VP of neurosecretory cells may be functionally related to the subarachnoid CSF and the arteriolar blood flow is discussed.
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  • 21
    ISSN: 1432-0878
    Keywords: ACTH cells ; Immunocytochemistry ; Morphometry ; Spontaneous hypertension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary ACTH-immunoreactive cells in the anterior pituitary of 4-week-old spontaneously hypertensive rats (SHR) were studied with immunocytochemical and morphometric techniques. The results were compared with data from age-matched normotensive Wistar-Kyoto rats (WKY). No significant differences were found in volume density and average size of ACTH-immunoreactive cells between these two strains. However, SHR showed a significantly larger anterior lobe (2 P 〈 0.01) than WKY, indicating that the total number of ACTH-immunoreactive cells in the anterior pituitary is greater in SHR than in WKY. These data are in agreement with radioimmunological determinations showing a significantly elevated content (2 P 〈 0.01) but only a moderately higher concentration (0.05 〈 2 P 〈 0.10) of ACTH in the anterior pituitary of SHR as compared to WKY. The present results suggest an enhanced availability of ACTH in the anterior pituitary of 4-week-old SHR, a fact which could explain the markedly enhanced stress-induced release of ACTH previously found in these animals. This study further supports the hypothesis that, among other factors, an instability of the hypothalamo-pituitary-adrenal axis may contribute to the development of genetically programmed hypertension.
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  • 22
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    Cell & tissue research 238 (1984), S. 289-296 
    ISSN: 1432-0878
    Keywords: Somatostatin (SRIF) ; Brain ; Pituitary gland ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical investigations show that somatostatin (SRIF)-like immunoreactive material is present in the brain and the pituitary of nine different species of teleosts. In the brain, immunoreactive perikarya and fibers are observed in the preoptic periventricular nucleus, the entopeduncular nucleus, the anterior periventricular nucleus, and the nucleus lateralis tuberis. In the pituitary, SRIF-like-immunoreactive fibers occur in the proximal pars distalis (PPD), which contains the growth hormone (GH)-secreting cells. Nerve fibers are scattered among GH cells (cyprinids), or end on the basal lamina at the neuroglandular interface of the PPD (eel, salmonids). In the eel, the proximal neurohypophysis does not penetrate deeply into the PPD that is very poorly vascularized. In some species, e.g. Myoxocephalus, SRIF-like immunoreactive fibers are also observed in the caudal neurohypophysis, and even among MSH cells of the pars intermedia. In long-term starved carps and eels, the amount of SRIF-like material in the pituitary is clearly reduced. A possible role of SRIF in the concomitant stimulation of GH cells is discussed.
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  • 23
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Slug brain ; Somatostatin-like material ; Maturation hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytological tests reveal the presence of a somatostatin-like substance in perikarya and axons in the brain of the giant slug Limax maximus L. Controls carried out on adjacent sections with absorbed antiserum or different antibodies raised against several biologically active peptides of vertebrates (ACTH-17-39, α- and β endorphin, α- and β MSH, methionin-enkephalin, TRH) demonstrate the specificity of the “staining”. However, some cells are both somatostatin- and FMRF-amide-positive. In the cerebral ganglia, the right Z-area cells, responsible for the synthesis of the maturation hormone (MH) are strongly somatostatin-positive. These results suggest a similarity between the MH and the somatostatin-like material contained in the Z-area cells. The simultaneous presence of two peptides in one and the same cell, the nature (elementary granules or soluble product) of the material, and its site of release are discussed.
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  • 24
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    Cell & tissue research 237 (1984), S. 195-202 
    ISSN: 1432-0878
    Keywords: Pituitary ; Prolactin cells ; Estrogen ; Heterogeneity ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study was conducted to determine the functional and/or developmental relationships among three heterogeneous types of prolactin cells (I, II and III) in rats. Rats were injected subcutaneously daily with estradiol or testosterone propionate on days 10–20 after birth. Estradiol increased the proportion of cell types II and III, increased serum PRL levels 12-fold in males and 15-fold in females, and increased pituitary levels of prolactin 12-fold in males and 5-fold in females. Testosterone mainly increased the proportion of the Type-II cells, decreased serum levels of prolactin in males only, and did not change pituitary levels of prolactin. In a second experiment, treatment of rats with nafoxidine for five days after E2 treatment (days 10–20 after birth) increased the proportion of Type-I cells and decreased the proportion of Type-III cells and decreased serum and pituitary levels of prolactin by 50% in females and by 15 and 45% in males. In a third experiment utilizing adult male rats, estradiol and testosterone were found to modulate the relative ratios of the different types of PRL cells as they did in immature animals. The data taken as a whole suggest the possibility of an estrogen-stimulated conversion of one cell type to another, which may be a reflection of prolactin secretory activity.
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  • 25
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    Cell & tissue research 237 (1984), S. 245-252 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Interstitial cells ; Astrocytes ; Immunocytochemistry ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected. It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.
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  • 26
    ISSN: 1432-0878
    Keywords: Crustacean hyperglycemic hormone ; Astacus leptodactylus ; Immunocytochemistry ; Quantitative electron microscopy ; Secretory cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical staining demonstrates striking differences in staining intensity among individual crustacean hyperglycemic hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus. Based on these differences we arbitrarily subdivided the CHH-cells into three categories representing increasing immunoreactivity respectively: + cells, + + cells, and + + + cells. Electron microscopic investigations reveal that these differences in immunostaining are correlated with differences in the numerical density of the neurosecretory granules in the cytoplasm and that these may reflect differences in activity among the CHH-cells. Morphometric analyses at the light- and electron-microscopic levels indicate that the three distinguished categories of immunopositive cells represent different stages in the CHH-synthesizing process of the cells. The results of the present study demonstrate the application of the PAP-technique at the light-microscopic level as a method to obtain information pertaining to the dynamics of secretory activity of the CHH-cells.
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  • 27
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    Cell & tissue research 238 (1984), S. 113-119 
    ISSN: 1432-0878
    Keywords: Crustacea ; Gastrointestinal hormones ; Neuropeptides ; Immunocytochemistry ; Cuticle ; Cancer magister
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A gastric peptide from the Dungeness crab (Cancer magister), extracted and characterized previously (Larson and Vigna 1983b), was localized in the foregut (stomach) of this species by immunocytochemistry using antisera specific for the bioactive carboxy-terminal amino acid sequence common to gastrins and cholecystokinins (CCKs). Immunoreactivity was found in all gastric epithelial cells and in the procuticle. Electron microscopy revealed an absence of peptidergic secretory granules in the gastric epithelial cells. The pattern of immunostaining suggests that the gastric epithelial cells secrete this peptide apically where it is incorporated into the cuticle lining the lumen. Specific immunostaining could not be demonstrated in various neural ganglia or in the hypodermis. The distribution of this peptide is different from that of gastrin/CCK in vertebrates and other invertebrates. This suggests that the crab gastric peptide is sufficiently similar to gastrin/CCK to react with C-terminal specific antisera, but may be anatomically, functionally, and possibly phylogenetically otherwise unrelated.
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  • 28
    ISSN: 1432-0878
    Keywords: FMRF-amide ; Immunocytochemistry ; Nervous system ; Microstomum lineare (Turbellaria)
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    Topics: Biology , Medicine
    Notes: Summary The central nervous system (CNS) and the peripheral nervous system (PNS) of the flatworm Microstomum lineare were studied by means of the peroxidase-antiperoxidase (PAP) immunocytochemical method, with the use of antisera to the molluscan cardioactive peptide FMRF-amide. FMRF-amide immunoreactive perikarya and nerve fibres are observed in the CNS and the PNS. In the CNS, immunoreactive perikarya and nerve fibres occur in the brain, in the epithelial lining and the mesenchymal surroundings of the ciliated pits, and positive fibres in the longitudinal nerve cords. In the PNS, immunoreactive fibre bundles with variocosities occur in the pharyngeal nerve ring, in symmetrical groups of perikarya on each side of the pharynx, and in the mouth area. Positive perikarya and meandering nerve fibres appear in the intestinal wall. A few immunoreactive cells and short nerve processes are observed at the male copulatory organ and on both sides of the vagina. Some immunoreactive peptidergic cells do not correspond to cells previously identified by histological techniques for neurosecretory cells. The distribution of immunoreactivity suggests that the FMRF-amide-like substance in CNS and PNS in this worm has roles similar to those of the brain-gut peptides in vertebrates. The status of FMRF-amide-like peptides as representatives of an evolutionarily old family of peptides is confirmed by the positive immunoreaction to anti-FMRF-amide in this primitive microturbellarian.
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  • 29
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    Cell & tissue research 238 (1984), S. 497-502 
    ISSN: 1432-0878
    Keywords: D2 glycoprotein ; Adrenal gland ; Immunocytochemistry ; Ultrastructure ; Cell adhesion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of the glycoprotein D2 in rat adrenal gland was investigated using immunohistochemical methods, and D2 localization in cultures of adult bovine chromaffin cells was studied by immunofluorescence. D2 was found to be situated on nerve fibers passing through the adrenal cortex and in the medulla zone, and also on the surface of all chromaffin cells. In addition, it was strongly expressed on the surface of glial (Schwann) cells. Cortical cells were unreactive to the antiserum. In cultures, all adrenalin and noradrenalin [dopamine-β-hydroxylase (DBH)-positive] cells were surface labelled for D2. A less frequent second cell type was recognized in vitro which was DBH negative but D2 positive. Such cells were presumed to be Schwann cells. These data are discussed in terms of the developmental origin of the cells and with regard to the putative functional rôle of D2 in cell adhesion phenomena.
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  • 30
    ISSN: 1432-0878
    Keywords: Retinal photoreceptors ; Opsin ; Optic nerve ; Immunocytochemistry ; Cyclostome, Myxine glutinosa
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    Topics: Biology , Medicine
    Notes: Summary Opsin-immunoreactive sites in the eye and optic nerve of the hagfish, Myxine glutinosa, were studied by use of light-microscopic pre- and postembedding peroxidase-antiperoxidase or avidin-biotin-peroxidase techniques, and the immuno-electron-microscopic protein A-gold method. At the light-microscopic level, a strong opsin immuno-reaction was obtained on the outer segments of the photoreceptor cells with sheep and rat antibodies against bovine (rhod)opsin. These outer segments were located in the marginal photoreceptor space and in follicles of the retina, as well as in the tubular lumen of the optic nerve. Ultrastructurally, two classes of outer segments can be distinguished; most of them exhibited a strong antiopsin reaction, while certain elements lacked immunoreactivity with the antisera employed. The protein A-gold particles marked opsin-immunoreactive sites on the photoreceptor membranes. The presence of opsin-immunoreactive material in the retina and optic nerve of the hagfish strengthens the view that this primitive eye lacking a cornea, lens and vitreous body is engaged in light perception. The morphological similarity between the eye and pineal tissue is discussed in connection with the absence of a pineal organ in this species.
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  • 31
    ISSN: 1432-0878
    Keywords: Pituitary rat ; LH cells ; FSH cells ; ACTH cells ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of immunocytochemistry, the development of ACTH-, LH- and FSH cells was examined in the anterior pituitary of 5-day-old neonatal, 15-day-old immature and adult rats. ACTH-positive cells are angular and the periphery of these cells is strongly reactive with anti-ACTH serum. In contrast, LH- and FSH-immunopositive cells are ovoid elements, ranging in cell size and intensity of staining. Angular cells, in which only the cell periphery reacted with anti-LHβ serum, were observed in neonatal and immature rats; however, these cells were not stained with either anti-FSHβ serum or anti-ACTH serum. Observation of serial semithin sections revealed that ACTH-immunopositive cells do not react with either anti-LHβ or anti-FSHβ serum. Finally, it was observed that ACTH cells and LH cells are both functionally differentiated already in 5-day-old neonatal rats.
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  • 32
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    Cell & tissue research 235 (1984), S. 77-83 
    ISSN: 1432-0878
    Keywords: Pituitary (rat) ; LH cells ; FSH cells ; Rapid freeze-substitution ; Immunocytochemistry ; Ferritin antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rapid freeze-substitution fixation was employed in immunocytochemical studies on the localization of LH and FSH in the typical gonadotrophs of the anterior pituitary in the untreated male rat; a modification of a recently described ferritin antibody method (Inoue et al. 1982) was used in these studies. It was shown that rapid freeze-substitution fixation provides good preservation not only of the ultrastructure but also of the antigenicity. Both LH and FSH were clearly demonstrated in the same gonadotrophic cells, but the subcellular localization of these gonadotrophins differed: (i) LH was mainly located in small secretory granules, 250–300 nm in diameter; (ii) FSH was mainly present in large secretory granules, up to 500 nm in diameter. In the pituitary gland of the adult male rat, all gonadotrophs that react to antibodies against gonadotrophins are characterized by small and large secretory granules. Other types of cells of the anterior pituitary containing either small secretory granules or resembling corticotrophs with secretory granules assembled at cell periphery did not react to either anti-LH beta or anti-FSH beta serum. For light microscopy, the peroxidase antibody method was used. All of the gonadotrophin-positive cells contain both LH and FSH. None of the pituitary cells reacted to antibody against only one gonadotrophin. However, some cells are “LH-rich” while other cells are “FSH-rich”.
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  • 33
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    Cell & tissue research 235 (1984), S. 159-169 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Gonadotropic cells ; Immunocytochemistry ; Testosterone binding ; Cryo-ultramicrotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several attempts have been made to localize steroids by means of immunocytological techniques. However, these methods were found inadequate for detecting steroids bound to their receptors. To localize endogenous testosterone (T) in its target cells at the ultrastructural level, an immunocytological technique was performed on ultrathin sections obtained by cryo-ultramicrotomy. T was detected in the pituitary glands obtained from intact male or female rats and castrated rats, but not in castrated + adrenalectomized rats. Animals were also injected either with testosterone, with other steroids (estradiol, progesterone, corticosterone) or with an androgen antagonist (cyproterone acetate). In addition, some ultrathin sections were preincubated either with phosphate buffers of various pH, corticosterone, cyproterone acetate solution, or with T solution. The content of T in the pituitary before and after fixation was measured by radioimmunoassay; it decreased after fixation. T immunoreactivity was localized in the gonadotropic cells only, both in the male and female rats. At the subcellular level, the immunoreactivity was detected in the cytoplasmic matrix and in the nucleus. Immunoreactive T disappeared 1) in rats after castration+adrenalectomy; by means of radioimmunoassay no T was measured in these pituitary glands; 2) in rats injected with 25 (μg/rat of cyproterone acetate; 3) after preincubation of pituitary sections on a drop of cyproterone acetate (1 × 10-6 M). The immunocytological reaction was not modified when the rats were injected with estradiol, progesterone or corticosterone (1 mg/rat), or after preincubation of the sections with corticosterone (1 × 10-3 M), or a buffer solution at pH 7.6. Lower or higher pH values led to a strong decrease in the immunoreactivity. After injection of T (15 μg/rat) the immunocytological reaction was more abundant in the nucleus and less in the cytoplasm. The immunoreactivity was again observed when the sections were preincubated with cyproterone acetate solution and then with T solution. These data suggest that T can be detected by means of immunocytochemistry. It is probably bound to a specific binding site.
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  • 34
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Supraoptic nucleus (SON) ; Oxytocin neurones ; Neuronal appositions
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    Notes: Summary Pre-embedding immunocytochemistry was performed on vibratome sections of the hypothalamus of lactating rats using antiserum directed against oxytocin. Electron microscopy revealed that numerous immunopositive somata and dendrites in the supraoptic nucleus were in direct apposition, without glial interposition; a number of them were also bridged by “double” synapses. The observations support the contention that the ultrastructural reorganisation of the nucleus apparent at lactation affects the magnocellular neurones secreting oxytocin.
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  • 35
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Goat ; Folliculo-stellate cell ; GH cell ; S-100 protein ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Goat pituitary glands were immunohistochemically studied with antisera for bovine S-100 protein, rat LHβ, FSH, TSHβ, prolactin, ovine GH, and porcine ACTH1–39 by use of the superimposition technique on adjacent sections. Folliculo-stellate (F-S) cells were divided into two categories on the basis of ultrastructural properties: One consisted of a mass of agranular cells in which the pseudolumina were equipped with microvilli and cilia. Elongate gap junctions were often observed among these cells. The other was a group of granulated cells with or without pseudolumina. In this group the gap junctions were shown to be disintegrated. The dense granules 150–250 nm in diameter began to accumulate in the cells. However, neither type of these F-S cells was immunostained for S-100 protein. On the other hand, numerous polygonal, elongate, irregular or stellate cells containing S-100 protein were distributed throughout the gland. Most of them were immunohistochemically identical with the GH cells laden with the secretory granules 250–450 nm in diameter, but some of them were identical to TSH and prolactin cells which immunostained faintly for S-100 protein. This appears to be the first demonstration of GH cells intensely immunostained for S-100 protein.
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  • 36
    ISSN: 1432-0878
    Keywords: Hagfish ; Brain ; Pituitary ; FMRF-amide ; Immunocytochemistry
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    Notes: Summary Paraffin sections of brain and pituitary of the hagfish Eptatretus burgeri were immunostained with an antiserum to FMRF-amide. Immunoreactivity was visible in a large number of neurons in the posterior part of the ventromedial hypothalamus and in long neuronal processes extending cranially from the hypothalamus to the olfactory system and caudally to the medulla oblongata. FMRF-amide-like immunoreactivity was also found in cells of the adenohypophysis. These observations suggest that the hagfish possesses a brain FMRF-amide-like transmitter system and pituitary cells containing FMRF-amide-like material. Antisera to ACTH, α-MSH and pancreatic polypeptide gave no immunoreaction in hagfish brain or pituitary.
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  • 37
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    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin interacting protein; Sobue et al. 1982) and actin was studied in the rat thyroid gland by means of light-microscopic immunocytochemistry, and the fine-structural distribution of actin filaments was examined by use of heavy meromyosin (HMM). Caldesmon and actin were demonstrated in the apical cytoplasm of almost all the follicle epithelial cells in normal as well as TSH-treated animals. Immunoreactivities for both caldesmon and actin showed almost the same pattern in localization. The smooth muscle cells of the blood vessels were also positive for caldesmon and actin. By electron microscopy, numerous actin filaments decorated by HMM and running perpendicularly or randomly to the apical surface were recognized in the apical cytoplasm of the follicle epithelial cell. These results suggest that caldesmon and actin, in conjugation with calmodulin, play a role in the regulation of cellular activity such as exocytosis and endocytosis in the apical portion of the follicle epithelial cell.
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  • 38
    ISSN: 1432-0878
    Keywords: Anterior pituitary, porcine ; Gonadotropic hormones (FSH, LH) ; Immunocytochemistry ; Cellular compartments
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    Notes: Summary Pre- and post-embedding immunocytochemical methods based on the use of specific antibodies against β-subunits of porcine LH and FSH were applied to determine the changes occurring in the anterior pituitary of the pig after gonadectomy. The results showed that (1) the total number of immunoreactive gonadotropes increased from 21–25% in control animals to 24–37% in castrated animals; (2) all gonadotropes contained both LH and FSH; (3) several types of immunoreactive LH/FSH cells were revealed; and (4) the two immunocytochemical methods used with dispersed cells localized the hormones in the same subcellular sites. However, the staining intensity in the different locations varied depending on the method applied. With the post-embedding method, a dense reaction product was found in the secretory granules but the cisternae of RER and the Golgi saccules were always slightly reactive. After the pre-embedding method, the staining intensity in the RER-cisternae and in the Golgi saccules was greatly increased. Thus, the two methodological approaches used in this study have permitted to visualize immunocytochemically the gonadotropic hormones not only at the sites of their storage but also along the intracellular pathway of the secretory material, i.e., at the site of its synthesis and during its passage via the Golgi zone.
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  • 39
    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor ; Immunocytochemistry ; Hypothalamus ; Domestic fowl
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    Topics: Biology , Medicine
    Notes: Summary The corticotropin-releasing factor (CRF)-containing neurons were investigated in the brain of the domestic fowl by means of the peroxidase-antiperoxidase technique at the light-microscopic level. The detection of CRF-immunoreactivity was facilitated by silver intensification. CRF-containing perikarya were found in the paraventricular, preoptic and mammillary nuclei of the hypothalamus and in some extrahypothalamic areas (nuclei dorsomedialis and dorsolateralis thalami, nucleus accumbens septi, lobus parolfactorius, periaqueductal gray of the mesencephalon, nucleus oculomotorius ventralis). Immunoreactive nerve fibers and terminals were demonstrated in the external zone of the median eminence and the organum vasculosum of the lamina terminalis. These results indicate that an immunologically demonstrable CRF-neurosecretory system also exists in the avian central nervous system.
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  • 40
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biogenic amines ; Vasotocin ; Lymnaea stagnalis
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    Notes: Summary Consecutive sections of certain neurons in the central ganglia of the pond snail Lymnaea stagnalis appear to be immunoreactive to anti-dopamine and anti-serotonin. The Cerebral Giant Neurons stain in addition with antivasotocin. The observations indicate the presence of two biogenic amines within the same neuron and in addition their co-existence with a biologically active peptide.
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  • 41
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    Cell & tissue research 237 (1984), S. 321-327 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Immunocytochemistry ; Respiratory tract ; Ear, middle
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    Topics: Biology , Medicine
    Notes: Summary Nerve fibres displaying neuropeptide-Y (NPY) immunoreactivity are abundantly distributed in the respiratory tract of cats, guinea-pigs, rats and mice. Fine beaded NPY fibres were seen in whole-mount spreads of the middle-ear mucosa. In the nasal mucosa and in the wall of the Eustachian tube NPY fibres were numerous around arteries and arterioles but sparse in the vicinity of veins; single fibres were found close to the acini of seromucous glands. In the tracheobronchial wall NPY fibres occurred in the proximity of blood vessels, in the subepithelial layer and in the smooth muscle. Surgical and chemical (6-hydroxydopamine treatment) sympathectomy resulted in disappearance of adrenergic and NPY-containing nerve fibres in the nasal mucosa. Sequential staining with antibodies against dopamine-β-hydroxylase (DBH) and NPY revealed that DBH and NPY occur in the same perivascular nerve fibres in the nasal mucosa. The distribution of NPY fibres in the respiratory tract suggests multiple functions of NPY, such as regulation of local blood flow, glandular secretion and smooth muscle activity.
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  • 42
    ISSN: 1432-041X
    Keywords: Vitellin ; Yolk granule ; Yolk protein ; Silkworm ; Embryogenesis ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Vitellin was purified from eggs of the silkworm,Bombyx mori, by a new method in which vitellin was extracted from isolated yolk granules. The purified vitellin had a molecular weight of 540,000. An antibody against purified vitellin was prepared in rabbits. It reacted with the hemolymph vitellogenin as well as with purified vitellin, but not with other proteins in the hemolymph or in the extract from yolk granules. The anti-vitellin IgG was used to immunocytochemically locate vitellin in theBombyx non-diapause egg during early developmental stages. In the egg, just after oviposition, vitellin was located in internal yolk granules and in small yolk granules of the periplasm. During the early developmental stages studied, vitellin was not metabolized uniformly throughout the egg. The vitellin of the internal yolk granules located at the posterior-dorsal part and of the small peripheral yolk granules was utilized in 16 h and 2 days, respectively, after oviposition. A thin, very vitellin-poor layer was located between the periplasm and the vitellin-rich interior in the newly laid egg. it was always in close contact with the periphery where blastoderm and germ-band cells developed.
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  • 43
    ISSN: 1432-0878
    Keywords: Pituitary ; Adenohypophysis ; Luteinizing hormone (LH) ; Immunocytochemistry ; Lamprey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A mammalian-like immunoreactive luteinizing hormone (LH) has been localized in the cytoplasm of cells in the meso-adenohypophysis of the sea lamprey, Petromyzon marinus L. Specific staining for LH was absent in other regions of the lamprey pituitary.
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  • 44
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    Cell & tissue research 229 (1983), S. 541-550 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Hagfish ; Somatostatin neurons ; Vasotocin neurons ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The brain of the Pacific hagfish, Eptatretus stouti, was studied immunocytochemically using antisera against somatostatin (SRIH), arginine vasopressin (AVP), and adrenocorticotropic hormone (ACTH). SRIH-immunoreactive perikarya were distributed bilaterally in the postoptic nucleus and in the hypothalamic nucleus. Although several short, stained fibers were observed in the vicinity of the perikarya, SRIH-immunoreactivity was not found in the neurohypophysis, nor in other parts of the brain. On the other hand, presumed arginine vasotocin (AVT) perikarya were distributed in an arc-shaped region extending from the posterior part of the preoptic nucleus to the anterior-most end of the hypothalamic nucleus and projected their fibers to the neurohypophysis. Most presumptive AVT perikarya were located close to the paired prehypophysial arteries near the anterior end of the postoptic nucleus. In the neurohypophysis, abundant presumptive AVT-fibers terminated in the posterior dorsal wall, although some fibers terminated in the anterior dorsal wall and only a few fiber endings were found in the ventral wall. No ACTH-positive cells were detected in the hagfish brain or in the pituitary gland.
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  • 45
    ISSN: 1432-0878
    Keywords: Somatotrops ; Oncorhynchus ; Immunocytochemistry ; Immunodiffusion
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    Topics: Biology , Medicine
    Notes: Summary Pituitary somatotrops were localized in four species from the genus Oncorhynchus using an antiserum to chum salmon (Oncorhynchus keta) growth hormone. The antiserum cross-reacted specifically with the acidophils in the caudal pars distalis of all species tested. Corroborative immunodiffusion studies were also conducted with pituitary homogenates from the same species. Immunological identity of the GH molecule was observed for all species tested.
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  • 46
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    Cell & tissue research 233 (1983), S. 99-111 
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Vasopression ; Osmotic stress ; RIA ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A correlative radioimmunoassay (RIA) and immunocytochemical (ICC) study was carried out on vasopressin (VP) distribution and content in brains of normal and 3-day water-deprived rats. By RIA there were statistically significant differences in brain VP per pg/mg between normal and osmotically stressed specimens in hypothalamus (338.4 versus 134.4), thalamus (4.8 versus 0.9), septum (18.0 versus 3.4), striatum (1.6 versus 0.7) and amygdala (17.3 versus 1.3), but not in other brain regions measured. Pituitary VP decreased from 71.1 to 8.7 ng/mg, and plasma VP rose from 3.6 to 19.3 pg/ml during water deprivation. Application of the peroxidase-anti-peroxidase ICC method of Sternberger to vibratome sections showed that VP-immunoreactivity in dehydrated specimens decreased in perikarya of paraventricular nucleus and suprachiasmatic nucleus, while intrahypothalamic immunoreactive magnocellular fibers appeared more conspicuous due to proliferation of large Herring bodies. In extrahypothalamic sites VP-immunoreactivity in water-deprived rats was visibly reduced in periventricular thalamus and septum. Thus it is apparent that both intra- and extrahypothalamic VP are affected by osmotic stress, and these results are discussed within the context of current ideas relating to co-activation of neurosecretory cells that project to different sites.
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  • 47
    ISSN: 1432-0878
    Keywords: Lung development ; Fibronectin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of the rat lung is a process of continuing morphological change. Indications from work in other mammalian systems suggest that fibronectin may be important in the control of this process. The present study has examined embryonic, neonatal, and adult lung tissue of the rat by means of the peroxidase-antiperoxidase (PAP) technique to demonstrate fibronectin at the light-microscopic level. Positive reaction was observed with anti-fibronectin serum in all stages examined. Control sections treated with pre-immune serum or no primary serum gave negative results in each case. Fibronectin in adult tissue was localized to the alveolar surface and alveolar basal lamina. Neonatal tissue showed fibronectin on pulmonary tubule walls and in basal lamina while embryonic tissue revealed localization of the protein in the basal lamina and in association with small groups of cells at the base of septal buds. These findings suggest a role for fibronectin in the control of rat lung development. The results are discussed in terms of the known functions of fibronectin as a preliminary matrix for the subsequent deposition of collagenous connective tissue, as a cellular adhesion protein, and as surface-bound material for cellular migration.
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  • 48
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    Cell & tissue research 233 (1983), S. 389-402 
    ISSN: 1432-0878
    Keywords: Calcium-binding protein (CaBP) ; Immunocytochemistry ; Rat cerebellum ; Development ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific antibodies raised against a human 28 000 dalton cerebellar calcium-binding protein (CaBP) were used in an immunocytochemical study during development of the rat cerebellum. Both light and electron microscopy showed (1) that labelling was entirely restricted to the Purkinje cells, (2) that it appeared very early in Purkinje cell development, (3) that the entire cell was labelled from the tip of the smallest dendrites to the axonal terminals, and (4) that with increasing age, the immunoreaction appeared to be progressively restricted to the cell and organelle membranes.
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  • 49
    ISSN: 1432-0878
    Keywords: Nucleus lateralis tuberis ; Hypothalamo-hypophysial relationship ; Immunocytochemistry ; Prolactin cells ; Teleosts
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    Topics: Biology , Medicine
    Notes: Summary In the rainbow trout the pars lateralis is the most prominent part of the nucleus lateralis tuberis (NLT). To demonstrate a morphological relationship between this lateral part of the NLT and the pituitary, immunocytochemistry was applied as a staining method. Experiments were carried out on glutaraldehyde-picric acid-acetic acidfixed brain sections of mature male and female rainbow trout using the peroxidase-anti-peroxidase immune technique with an antiserum against 27-S-methylglucagon as the first antibody. Most of the cells in the NLT/pars lateralis reacted with the antiserum. Axons from these cells enter the pituitary, extending exclusively in the numerous neurohypophysial digitations in the pars intermedia. No immunoreactive neurohypophysial protrusions were found in those parts of the adenohypophysis where the gonadotropic cells are located, indicating that the lateral part of the NLT is not directly involved in the control of gonadotropin secretion. In addition to cells of the NLT/pars lateralis only prolactin cells in the rostral pars distalis of the adenohypophysis reacted with the antiserum used.
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  • 50
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    Cell & tissue research 228 (1983), S. 525-534 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasopressin ; Oxytocin ; Bed nucleus, stria terminalis ; Lateral septum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the dorsal and ventral portions of the bed nucleus of the stria terminalis of the rat numerous cell bodies immunoreactive for vasopressin and neurophysin II were found after colchicin pretreatment. These cells are predominantly multipolar but sometimes also bipolar, and have a width and length of approximately 9 and 16 μm, respectively. In the homozygous Brattleboro rat, which is deficient in vasopressin, no immunoreactive vasopressin was found in these cells. Following incubation with anti-oxytocin and anti-bovine neurophysin I, only magnocellular immunoreactive cell bodies were found in the septal region. The consequences of these results concerning the vasopressin fiber pathways in the brain are discussed.
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  • 51
    ISSN: 1432-0878
    Keywords: TRH ; Immunocytochemistry ; Autoradiography ; Arcuate nucleus ; Median eminence ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical and autoradiographic localization of thyrotropin-releasing hormone (TRH)-and 3H-TRH-binding sites was studied in the arcuate nucleus-median eminence region of the rat. TRH-like immunoreactivity was found in dense granular vesicles (90–140 nm in diameter) in TRH-like immunoreactive nerve fibers and terminals. In the median eminence, the immunoreactive terminals were observed to be in direct contact with the perivascular basal lamina of the portal vessel and to form synaptoid contacts with tanycytes. In the arcuate nucleus, the immunoreactive terminals were often found to form axosomatic and axo-axonic, and/or axo-dendritic synapses. The uptake of tritiated TRH into the nerve fibers and terminals of the median eminence was also observed by autoradiography and the distribution and localization of silver grains in them were analyzed quantitatively by circle analysis. Thirty minutes after intraventricular infusion of 3H-TRH, radioactive labeling occurred in type-2 and 3-nerve fibers and terminals containing dense granular vesicles in the median eminence. It is therefore suggested that the neurons labeled after 3H-TRH infusion possess certain functions as physiological recognition sites or receptors for TRH.
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  • 52
    ISSN: 1432-0878
    Keywords: Adipokinetic hormone ; Locusta migratoria ; Neurohormone ; Peptidergic neuron ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum was raised against tyrosine-adipokinetic hormone ([Tyr1]-AKH). In immunohistochemical procedures, it revealed the AKH cells in the glandular lobes of the corpora cardiaca (CC) of Locusta migratoria with high specificity. In addition, an immunologically related peptide was detected in certain neurons of the central nervous system which suggests that this AKH-like peptide may have a neurotransmitter function. The glandular lobes contain immunoreactive AKH cells in all postembryonic stages, and no essential differences in morphology and distribution of the cells in nymphs and adults were seen. The amount of AKH, stored predominantly in the cell projections, differs widely among cells and individuals. The brain of adults and nymphs contains several small populations of intensely stained neurons. In last-instar and adult specimens, each half contains 10–12 “normal”-sized neurons in the protocerebrum (including the optic lobe) and deutocerebrum, and in addition 15–18 small reactive neurons. Their axons and numerous branchings traverse the neuropile of proto-, deuto-, and tritocerebrum, except for the pedunculate bodies and antennal lobes. Some of the axons run into the storage lobe of the CC; it is unknown if their content is released into the haemolymph. Other axons run into the ganglia of the stomatogastric nervous system and into the circumoesophageal connectives. The suboesophageal ganglion also contains 8 immunoreactive neurons. It is unknown to which extent the immunoreactive substances in glandular and nervous tissue are chemically and physiologically related.
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  • 53
    ISSN: 1432-0878
    Keywords: Thyroid ; Pituitary ; Thyrotrops ; Gonadotrops ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Alcian blue and periodic acid-Schiff-positive “granular basophils” with electron-dense granules ±160 nm in diameter, and weakly developed irregular cisternae of the granular endoplasmic reticulum (GER) are present in the rostral pars distalis (RPD) and the proximal pars distalis (PPD) of immature rainbow trout. They all react with antisalmon αβ-gonadotropin. However, only granular basophils in the caudal RPD and the rostro-dorsal PPD, often bordering on and sometimes surrounded by neurohypophysial tissue, react with anti-human β-TSH. These cells, considered as the source of thyrotropin, show degranulation and dilatation of the GER-cisternae in fish treated with potassium perchlorate. The thyroids of the goitrogen-treated animals had relatively numerous small follicles with a high epithelium. The remaining granular basophils are gonadotrops.
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  • 54
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    Cell & tissue research 233 (1983), S. 233-236 
    ISSN: 1432-0878
    Keywords: Y-organ ; Steroid secretion ; Ecdysone ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary Ecdysone was demonstrated by ultrastructural immunocytochemistry to be present in the mitochondria of the Y-organs of the crayfish Orconectes limosus. This is in remarkable contrast to the prothoracic glands of insects and suggests substantial differences in the biosynthesis of the same hormone, ecdysone, in crustaceans and insects.
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  • 55
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    Cell & tissue research 234 (1983), S. 427-437 
    ISSN: 1432-0878
    Keywords: Corticotropes ; Rat fetus ; Ultrastructure ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary Corticotropes of rat fetuses aged 16, 18 and 21 days were localized by the indirect antibody-enzyme method on semithin sections of the pituitary. The development of the ultrastructure of these cells was observed on consecutive ultrathin sections. In comparison with previous data our present results show that identification of a fetal cell type cannot be based entirely on morphological criteria. The structural peculiarities of corticotropes obtained from studies in vivo are compared with those observed in cells maintained in vitro.
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  • 56
    ISSN: 1432-0878
    Keywords: Villin ; Intestinal brush border ; Contractile proteins ; Immunocytochemistry ; Immunostaining of blotted gels
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    Topics: Biology , Medicine
    Notes: Summary Villin, a 95,000 dalton polypeptide of intestinal brush border which is known to bundle or sever actin filaments in a Ca++-dependent manner, was localized in rat and chicken intestinal epithelium by means of immunocytochemistry at the light- and electron-microscopic levels. Specific antibodies to villin were raised in rabbits immunized with villin purified from chicken intestinal epithelium. Anti-villin bound selectively to the microvillus filament bundle from its tip down to the rootlets. These findings indicate that the well-known stability of rootlet filaments towards elevated Ca++ ion concentrations cannot be explained by the absence of villin. Therefore additional factors must exist which prevent the rootlets from Ca++-villin mediated disassembly.
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  • 57
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    Cell & tissue research 231 (1983), S. 205-213 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Insulin ; Glucagon ; Pancreatic polypeptide ; Diabetes mellitus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rats rendered diabetic by streptozotocin were subjected to pancreas transplantation. After twenty weeks, the duct-ligated pancreas transplant was studied morphometrically to determine the effect of duct occlusion on the various cell populations of the islets. Concomitantly, the streptozotocin-treated host pancreas was examined for a possible influence of the graft on the diabetic pattern of islet cell population. Twenty weeks after pancreas transplantation, the volume fractions of insulin, glucagon, somatostatin and pancreatic polypeptide cells in the graft islets did not differ from those of the normal control pancreas. In the pancreas of nontransplanted diabetic rats, insulin-positive B cells were reduced from 60–65% to less than 10% of the islet volume, whereas non-B cells were significantly increased in volume density. The changes in fractional volume of the various islet cells correlated fairly well with changes in plasma concentration of the corresponding pancreas hormones. In the recipient's own pancreas, the relative volumes of glucagon and somatostatin cells were unaffected by the pancreas transplant. However, the insulin cell mass was significantly increased, and comprised about 20% of the islet volume, while cells containing pancreatic polypeptide were found only sporadically.
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  • 58
    ISSN: 1432-0878
    Keywords: Pituitary gland ; S-100 protein ; Castration ; Thyroidectomy ; Postnatal development ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary folliculo-stellate cells and associated cells were studied immunohistochemically throughout the life of male and female rats. The marginal layer cells of the pars distalis and intermedia as well as the folliculo-stellate cells were immunostained with anti-S 100 protein serum. The immunostained folliculo-stellate cells in one section were identified as themselves by their ultrastructural properties on the adjacent section. Corticotrophs, thyrotrophs and prolactin cells, characterized by the stellate shape, were not immunostained with anti-S 100 protein serum. Reactivity for S-100 protein appeared on postnatal Day 6 in the marginal layer cells of the pars intermedia, and appeared on Day 10 in those cells of the pars distalis. No immunostained folliculostellate cells appeared before Day 6, but a few of them appeared on Day 10. Thereafter they increased in number, having more intense immunostaining with advancing age. In castrated rats, the immunostained folliculo-stellate cells appeared quite numerously, with branched cytoplasmic processes surrounding the gonadotrophs. In the thyroidectomized rats, however, folliculo-stellate cells lacked ramified cytoplasmic processes, and their topographic affinity for thyrotrophs was negligible.
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  • 59
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    Cell & tissue research 231 (1983), S. 527-532 
    ISSN: 1432-0878
    Keywords: S-100 ; Müller cells ; Immunocytochemistry
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    Notes: Summary The localization of S-100 protein in the retina was studied by immuno-electron microscopy. The immunoreaction for S-100 is specifically confined to Müller cells and astrocytes. Long slender cytoplasmic processes of Müller cells and their participation in the formation of the inner and outer limiting membrane were clearly demonstrated with this immunoreaction. The cytoplasmic processes were also revealed to form continuous sheets surrounding blood vessels. Some somata of Müller cells were found in the outer nuclear layer. The results show that immunostaining for S-100 is useful for selective visualization of Müller cells in the mature retina.
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  • 60
    ISSN: 1432-0878
    Keywords: Baboon ; Pituitary gland ; Gonadotrophs ; Mammotrophs ; Somatotrophs ; Thyrotrophs ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary The identification and regionalization of four pituitary parenchymal cell types, gonadotrophs, mammotrophs, somatotrophs and thyrotrophs, were studied in the baboon (Papio cynocephalus) hypophysis using immunocytochemistry. The gonadotrophs were homogeneously distributed throughout the entire pars distalis. Both mammotrophs and somatotrophs predominate at the superior and inferior poles of the organ. The medial and anteromedial regions are populated by mammotrophs and thyrotrophs, while the lateral and posterior portions of the pars distalis contain large numbers of somatotrophs.
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  • 61
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Human TSH β subunit ; Teleosts ; TSH cells
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    Topics: Biology , Medicine
    Notes: Summary A rabbit antiserum raised against human thyrotropin (TSH) β subunit (anti-human β-TSH) was used to identify TSH cells in the pituitaries of five species of teleosts by means of the unlabeled antibody peroxidase-antiperoxidase (PAP) technique. The anti-human β-TSH reacted only with polygonal basophils located in the rostral pars distalis (RPD) in the pituitaries of sexually immature eel (Anguilla japonica). In the pituitaries of medaka (Oryzias latipes) and white-edged rockfish (Sebastes taczanowskii), this serum reacted specifically with only one cell-type located in the antero-dorsal proximal pars distalis (PPD) and antero-ventral PPD, respectively. In the pituitaries of two species of salmonids, sexually immature rainbow trout (Salmo gairdneri) and sexually mature chum salmon (Oncorhynchus keta), the anti-human β-TSH bound to the polygonal basophils in the ventral PPD adjacent to the RPD. Thus, the present immunocytochemical study identified polygonal basophils located in the RPD or PPD as TSH cells in five species of teleosts.
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  • 62
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    Cell & tissue research 232 (1983), S. 129-140 
    ISSN: 1432-0878
    Keywords: Serotonin ; Optic lobes ; Insecta ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular localization of 5-HT in the optic lobes of three insect species was assayed with the use of antibodies raised against 5-HT. In Schistocerca, Periplaneta, and Calliphora all neuropil regions of the optic lobe, the lamina, medulla and lobula, contain 5-HT-immunoreactive varicose fibres in different patterns, like columns and layers. Such fibres also connect the lobula to neuropil in the lateral protocerebrum. In Calliphora also 5-HT-positive fibres of the medulla and lobula plate have projections to the lateral protocerebrum, whereas the origin of the lamina fibres is not certain. In all species the processes displaying 5-HT-like immunoreactivity appear to be derived from a relatively small number of cell bodies, each neuron thus having processes over a large volume of the neuropil of the optic lobe in different layers.
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  • 63
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    Cell & tissue research 234 (1983), S. 547-559 
    ISSN: 1432-0878
    Keywords: Myelin proteolipids ; Oligodendrocytes ; Golgi apparatus ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Purified antibodies directed against myelin proteolipids were isolated by affinity chromatography of whole serum obtained from rabbits inoculated with myelin. These antibodies were specific for light, medium and dark oligodendrocytes. Astrocytes, neurons and their processes were not reactive. Immunocytochemical investigations showed that the membranes of the Golgi complex are highly labeled by these antibodies. Diffuse cytoplasmic labeling was only observed on the light and medium oligodendrocytes and was absent from the dark types. Vesicles possessing a punctate staining were detected in the vicinity of the Golgi complex and the oligodendroglial membrane. A discontinuous labeling of the plasmalemma appears to be characteristic of the actively myelinating light and medium oligodendrocytes. In compact myelin sheaths positive immunostaining was only detected at the dense line. The immunocytochemical localization of the myelin proteolipids in the oligodendrocytes is in accordance with previously published biochemical data.
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  • 64
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    Cell & tissue research 233 (1983), S. 69-80 
    ISSN: 1432-0878
    Keywords: Somatostatin ; Hypothalamus ; Immunocytochemistry ; Human ; Rhesus monkey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical methods were used to compare the localization of somatostatin (SRIF) in the human and rhesus monkey hypothalamus. The distribution of SRIF-containing cell bodies and fibers is similar in the two species. Perikarya are located predominantly in the periventricular region and to a lesser extent in the ventromedial nucleus. Fibers occur in dense clusters within the periventricular region, ventromedial nucleus, arcuate nucleus, median eminence, and pericommissural area of both species. Analysis of serial sections suggests that fibers originate from cells in the periventricular region, extend ventrally through the ventromedial and arcuate nuclei to terminate around the portal vessels of the infundibular stalk, and thereby participate in the regulation of anterior pituitary function. Somatostatinergic fibers are also found surrounding non-immunoreactive perikarya in the ventromedial nucleus and periventricular region of both primates. This arrangement may support somatostatin's postulated role as a neurotransmitter or neuromodulator. The strong similarity between the localization of hypothalamic SRIF in the human and rhesus monkey supports the use of the rhesus monkey as a model for the study of somatostatin as a neuroendocrine regulator in the human.
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  • 65
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasopressin ; Dorsomedial hypothalamus ; Amygdala ; Locus coeruleus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Recently, the existence of a vasopressin-immunoreactive cell group was described in the bed nucleus of the stria terminalis (van Leeuwen and Caffé 1983). In the present investigation additional nuclei containing vasopressin-immunoreactive cells were found, after colchicine pretreatment, in the dorsomedial hypothalamus, medial amygdaloid nucleus and the locus coeruleus. Vasopressin-immunoreactive cells in the dorsomedial hypothalamus and medial amygdaloid nucleus are small (8–14 μm and 10–14 μm, respectively), while those in the locus coeruleus are medium-sized (20–25 μm). Incubation with anti-bovine neurophysin II and anti-rat neurophysin revealed staining of the same cell group in the above-mentioned areas. None of these cell groups show stained cells after incubation with anti-oxytocin and anti-bovine neurophysin I. When sections of the homozygous Brattleboro rat, which shows a deficiency in vasopressin synthesis, are incubated with anti-vasopressin, anti-bovine neurophysin II, or anti-rat neurophysin, no immunoreactivity can be observed in these brain regions. The above-mentioned cell groups may contribute to the vasopressinergic innervation of brain sites that have been reported to persist after lesioning of the suprachiasmatic, paraventricular and bed nuclei of the stria terminalis.
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  • 66
    ISSN: 1432-0878
    Keywords: α-Endorphin ; Immunocytochemistry ; Median neurosecretory cells ; Peptidergic neurones ; Calliphora vomitoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A group of the 24–26 paraldehyde fuchsin-positive median neurosecretory cells (MNC) in the pars intercerebralis of the brain of the blowfly, Calliphora vomitoria, has shown immunoreactivity towards three different antibodies to α-endorphin, a peptide that corresponds to the amino acid sequence present between residues 61 and 76 of the precursor molecule, β-lipotropin (β-LPH). The immunoreactive material could be followed in axons within the median bundle, the tract through which neurosecretory material from the MNC is passed down to the corpus cardiacum (CC). The α-endorphin-immunoreactive material was observed leaving the CC in the cardiac-recurrent nerve, dorsal to the proventriculus, in the direction of the abdomen. The cells that contain the α-endorphin-like material are different from those of the MNC that contain insulin-, pancreatic polypeptide-, and gastrin/CCK-like peptides. This finding demonstrates the considerable complexity and peptidergic nature of the MNC and constitutes further evidence that morphinomimetic-like peptides are present in the nervous system of invertebrates.
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  • 67
    ISSN: 1432-0878
    Keywords: Angiotensinogen ; Liver ; Kidney ; Immunocytochemistry
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    Notes: Summary The renin substrate, angiotensinogen, was localized by immunocytochemistry in liver and kidney of normal rats by the use of an antiserum directed against pure rat angiotensinogen. This substrate was also examined in rats after bilateral nephrectomy, which is known to increase plasma angiotensinogen, and in rats treated with colchicine, which inhibits serum protein secretion. In normal rat liver, light microscopy showed the presence of immunoreactive material in a very few cells. The number of stained hepatocytes rose in rats treated with colchicine or after bilateral nephrectomy. Immuno-staining increased further when rats were both nephrectomized and colchicine treated. In the kidney, angiotensinogen was specifically located as granular formations in nephrocytes of the proximal tubule but never in the granular cells of the juxtaglomerular apparatus. The localization of these granular formations under the brush border suggests that angiotensinogen is reabsorbed from the glomerular ultrafiltrate rather than synthesized in the kidney.
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  • 68
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    Cell & tissue research 233 (1983), S. 471-474 
    ISSN: 1432-0878
    Keywords: Prolactinergic neurons ; Immunocytochemistry ; Cerebral ganglion ; Styela plicata
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    Topics: Biology , Medicine
    Notes: Summary Peptidergic neurons have been detected in the cerebral ganglion of the ascidian Styela plicata by means of cytochemical methods. After incubation with a mammalian antibody to human prolactin the perikarya show a strong immunoreactivity. The possible function of prolactin-like peptides in the nervous system of protochordates is discussed.
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  • 69
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    Cell & tissue research 234 (1983), S. 125-134 
    ISSN: 1432-0878
    Keywords: Vasopressin ; Immunocytochemistry ; Electron microscopy ; Hypothalamus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vasopressin-containing neurons, identified by immunocytochemistry, are located predominantly in the posterior magnocellular division of the paraventricular nucleus of the rat hypothalamus. By electron microscopy, the immunoreaction product is seen within the cell bodies and neuronal processes. In the perikarya and dendritic processes, the immunoreactive material is associated primarily with neurosecretory granules. Axonal processes, identified by their content of microtubules and accumulation of neurosecretory granules, show the immunoreaction product in association with both of these organelles. Afferent axo-dendritic, axo-somatic and putative axo-axonic synapses with immunostained vasopressinergic neurons can be identified. The presynaptic profiles do not contain immunoreactive material. This study contributes to the ultrastructural characterization of vasopressinergic neurons in the paraventricular nucleus and of their afferent synaptic input.
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  • 70
    ISSN: 1432-0878
    Keywords: Fibronectin ; Lung ; Development ; Ultrastructure ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In a previous study changes in the macrodistribution of fibronectin during rat-lung development were examined. Using the peroxidase-antiperoxidase immunocytochemical technique, we have demonstrated the presence of fibronectin in embryonic, neonatal, and adult rat lung at the ultrastructural level. In the embryo, fibronectin is found both in an intra-and extracellular association with isolated pneumoblasts, and in a periodic distribution along the basal lamina. The neonate displays fibronectin in an intracellular association with early type-I cells and on their basal and luminal surfaces, but not in association with type-II cells. Neonatal basal lamina is diffusely labeled by anti-fibronectin antiserum. Fibronectin in adult tissue is found both intracellularly and on the basal and luminal surfaces of type-I cells but not in type-II cells. The basal lamina and interstitial connective tissue are slightly or non-reactive. These observations confirm and extend our initial suggestion that fibronectin is involved in rat-lung development.
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  • 71
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Uteroglobin ; Rabbit ; Epididymis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the immunoperoxidase technique and specific goat antiserum to uteroglobin, selective immunochemical staining products are localized in the epididymal epithelium of the caput and proximal corpus region, at the adluminal border of the cauda epididymidis and, as well known, in epithelial cells of the endometrium of pregnant and progesterone-treated rabbits. Specific staining is also seen on spermatozoa. A uteroglobin-like antigen has been similarly localized in alveolar and bronchial epithelial cells of the lung. Testis, prostate, seminal vesicle and ductus deferens do not seem to contain in their tissues immunoreactive uteroglobin-like antigens. Similarly, the uterus and ductus epididymidis of immature rabbits are devoid of immunoreactivity. The presence of uteroglobin-like antigens in tissues other than the endometrium, particularly the ductus epididymidis, stimulates new discussions on the function of this protein in reproductive physiology and fertility research.
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  • 72
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ontogeny ; LHRH ; Brain ; Teleost fish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have traced the development from birth to puberty of the ir-LHRH producing centers, the nucleus olfactoretinalis (NOR), nucleus praeopticus periventricularis (NPP) and nucleus lateralis tuberis pars posterior (NLT), in male sibling platyfish genetically determined by differing P alleles to mature at two different ages. In sexually immature fish of both genotypes ir-LHRH is first localized in the NOR in perikarya and neuronal fibers and in the NPP where it is limited to the fibers. At the onset of puberty in early maturing fish (11 weeks), ir-LHRH accumulates in perikarya and fibers of the NPP and NLT. Late maturing siblings first have ir-perikarya in the NPP and a few ir-fibers in the NLT at the same developmental stage but at a later age (25–26 weeks). Ir-perikarya are still not seen in the NLT in late-maturing fish even when maturation is complete. This pattern of a sequential development of the three ir-LHRH containing areas of the brain, which we refer to as a “cascade effect”, is similar in early and late genotypes. It is directly correlated to stage of development and not to chronological age. It is clear that the appearance of LHRH producing brain centers precedes, and is presumably essential for, the completion of the development of pituitary gonadotrops and the subsequent maturation of the gonads. The functional significance of the NOR in puberty and reproduction and as a possible site of action for the P gene is discussed.
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  • 73
    ISSN: 1432-0878
    Keywords: Pars intermedia ; Oxytocin ; Dopamine ; Serotonin ; Immunocytochemistry ; Rabbit, hare
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pars intermedia of the pituitary in the rabbit and hare is abundantly innervated by axons reacting selectively with antibodies against oxytocin. These axons contain dense secretory vesicles about 140 nm in diameter, i.e., smaller than those in the neurosecretory axons of the neural lobe. No fiber elements staining for other peptides (vasopressin, somatostatin, substance P) were observed in the pars intermedia, except rare leu-enkephalin axons restricted to the rostral zone of the gland. Dopaminergic innervation appears to be completely absent from the intermediate lobe. This was shown by the lack of reaction with an antibody against tyrosine-hydroxylase, which did reveal a well-developed tubero-infundibular system of nerve fibers. Axons reacting with an antibody against serotonin were irregularly distributed in the pars intermedia. In the absence of dopaminergic axons, the extensive oxytocin-like innervation may play a major role in regulating the melanotrophic cell activity in the Leporidae.
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  • 74
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    Cell & tissue research 228 (1983), S. 265-276 
    ISSN: 1432-0878
    Keywords: Met- and leu-enkephalin ; Immunocytochemistry ; Ultrastructure ; Lateral septum ; Guinea-pig brain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By the use of antisera to met-enkephalin and leu-enkephalin, enkephalin-containing structures were visualized in the lateral septum of the guinea-pig brain. The present results do not reveal immunoreactive perikarya in this area. The immunostaining is exclusively located in numerous nerve fibers and endings mostly encompassing neuronal perikarya, which accounts for the fact that at the light-microscopic level cellular somata appear to be immunostained. The immunoreactive terminals and fibers contain granules approximately 110nm in diameter and synaptic vesicles. The origin and the functional role of these numerous enkephalin terminals remain to be established. This work was supported by a grant from INSERM N∘ 804012
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  • 75
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    Cell & tissue research 234 (1983), S. 319-333 
    ISSN: 1432-0878
    Keywords: Serotonin ; Goldfish ; Immunocytochemistry ; Central nervous system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of immunoreactive serotonin (5-HT) was investigated in the brain and pituitary gland of the goldfish with the use of indirect immunofluorescence technique. Immunoreactive cerebrospinal fluid-contacting neurons were found in the nucleus recessus lateralis and in the nucleus recessus posterions of the hypothalamus. Additional immunoreactive neurons were observed ventro-lateral to the posterior commissure in the nucleus dorsolateralis thalami. This group of cell bodies extends posteriorly as far as the rostral midbrain tegmentum. At the level of the isthmus, numerous immunoreactive cell bodies were located medially between the fiber bundles of the fasciculus longitudinalis medialis. Finally, a few isolated immunoreactive cells were observed in the medulla oblongata. In the pituitary gland, immunoreactive fibers and cells were found primarily in the pars distalis. The origin of the numerous fibers of the telencephalon is not clear, however, they may arise from the perikarya located in the raphe region. The general organization of the ascending and descending 5-HT-systems of the teleost brain appears to be similar to that described in mammals.
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  • 76
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    Cell & tissue research 234 (1983), S. 439-450 
    ISSN: 1432-0878
    Keywords: Avidin ; Avidin-biotin interaction ; Biotin affinity histochemistry ; Biotin hydrazide ; Immunocytochemistry ; Magnum gland ; Secretion ; Oviduct
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of endogenous avidin was studied cytochemically in the magnum tissue of the oviduct of laying hens. Two methods, based on an interaction of avidin-biotin with biotin hydrazide-peroxidase (B-HRP) as an affinity reagent, and on an immunoperoxidase technique, were tested by morphological analysis. The data obtained by both methods showed that in the magnum B-HRP is a strictly substitutive reagent for endogenous avidin. Avidin was clearly demonstrated in large amounts in the secretory granules of some epithelial cells and tubular gland cells, but was absent from mucous cells, the goblet cells, which had been believed to be the location of avidin production, and from ciliated cells. These granules had previously been demonstrated by both electron-microscopic cytochemical techniques. Especially in acinar cells, they were nonhomogeneous with a speckled core and a dense peripheral part. They ranged in size from 500 to 2200nm in diameter in the gland and 180 to 720 nm in the epithelium. Columnar epithelial cells containing avidin granules had a strong resemblance to those of the protodifferentiated tubular gland cells in the magnum of chicks pretreated with daily estrogen or estrogen plus progesterone, and might have migrated towards the acinus as substitutional secretory cells. Therefore, the acinar cells of the magnum, considered to be composed of several secretory protein-producing systems, are dependent on estrogen and/or progesterone in the oviduct of the laying hen.
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  • 77
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    Cell & tissue research 222 (1982), S. 81-100 
    ISSN: 1432-0878
    Keywords: Turtle ; Immunocytochemistry ; Pars tuberalis ; Pituitary ; Hormones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Examination of pituitaries from young and adult turtles representing four families, reveals that in addition to the abundant juxtaneural pars tuberalis (JuxPT) found in this class of reptiles, there is generally a substantial amount of pars tuberalis (PT) tissue closely associated with the pars distalis (PD). The PT forms a cortical layer especially conspicuous around the anterior tip of the PD in some species (Trionyx, Kinosternon, Sternotherus), or it forms a thick dorsal layer of tissue irregularly extending onto the sides of the PD in others (Pseudemys, Chrysemys, Lepidochelys, Chelonia). Immunocytochemical studies using unlabelled second antibody and peroxidase-antiperoxidase reveal that in turtles of all ages, the PT tissue allied with the PD (the PTinterna) is composed primarily of cells containing glycoprotein hormones (FSH, LH and TSH), especially the gonadotropins. The juxPT, however, consists mainly of secretory cells unstained by the antisera tested and includes only a small number of gonadotropes and thyrotropes. Although usually widely distributed in the testudinate adenohypophysis, the great majority of gonadotropes and thyrotropes present in the hatchling are in the PTinterna. It is probable that a concentration of these cells in the PTinterna is widespread among vertebrates. In all turtles examined, lactotropes occur principally in the anterior and ventral part of the PD proper; somatotropes are posterior and dorsal. Corticotropes are concentrated as the lactotropes in the anterior PD, but some are also scattered throughout the posterior half of the gland. Lactotropes, corticotropes, and with a few exceptions, somatotropes, do not occur in PT tissue in the turtle.
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  • 78
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    Cell & tissue research 223 (1982), S. 287-303 
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Avian brain ; Endocrine glands ; Avian embryos ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In embryos of the domestic mallard, domestic fowl, and Japanese quail vasotocin-, mesotocin-, luliberin (LHRH)-, met-enkephalin-, cortico- tropin-, and somatostatin-immunoreactive perikarya and fiber formations were visualized at different incubation stages by means of the PAP technique (Sternberger 1979). The most striking results were: (1) Vasotocin-, mesotocin-, and luliberin-immunoreactive systems display, up to the late embryonic period, morphological features most probably related to a neurohormonal function. (2) Met-enkephalin immunoreactivity appears very late during embryonic life; it is restricted to fiber networks and not found in perikarya. (3) Corticotropin immunoreactivity is observed in the tuberal region temporarily at the end of the second and the beginning of the last third of the incubation period. (4) Somatostatin-immunoreactive material is present (i) at the end of the first third of incubation, in association with the olfactory system; (ii) during the same period, adjacent to thin-layered portions of the roof of the brain; (iii) shortly thereafter, in cells of both pancreatic primordia and thyroid gland; and (iv) onward from the middle of the incubation period, in a mesencephalic cell group. The striking difference, in the early embryo, between the mature somatostatin system and the immature character of the surrounding tissues may indicate that somatostatin plays a role in the development of the brain, as well as the pancreas, and the thyroid gland.
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  • 79
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    Cell & tissue research 223 (1982), S. 695-698 
    ISSN: 1432-0878
    Keywords: Pineal gland ; Luteinizing hormone-releasing hormone (LHRH) ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactive luteinizing hormone-releasing hormone (LHRH)-like material has been demonstrated in the pineal gland of the adult rat. The objective of the present study was to examine the ontogenetic development of this LHRH-like substance in the rat pineal with the peroxidase-antiperoxidase (PAP) method of Sternberger. LHRH-like immunoreactive material was not observed in pineal glands of newborn rats. The amount of material increased progressively from the 6th–12th day of postnatal development. On day 12, the amount of LHRH-like immunoreactivity was consistent and comparable in all pineal glands of male and female animals examined.
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  • 80
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    Cell & tissue research 224 (1982), S. 25-31 
    ISSN: 1432-0878
    Keywords: Spleen ; Ig-containing cells ; Immunocytochemistry ; Germinal center ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphology and localisation of IgM- and IgG-containing cells in the spleen of rats immunized with sheep red-blood cells (SRBC) were studied by combining immunohistochemical reactions with routine histological and histochemical methods. It was shown that IgM cells occur only in the outer periarteriolar lymphocyte sheath (PALS), whereas IgG cells are present throughout the whole PALS. It has been concluded that these cell types have different routes of migration. Both IgM- and IgG-containing cells were found in the germinal center in close relation with the extracellular immune complexes. The significance of this localisation is discussed.
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  • 81
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    Cell & tissue research 224 (1982), S. 361-368 
    ISSN: 1432-0878
    Keywords: Steroids ; Pituitary ; Gonadotrops ; Immunocytochemistry ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical reactions with anti-h(uman) TSHβ and anti-c(arp) GTHβ indicate that in juvenile rainbow trout, the dorsal basophils of the proximal pars distalis (ppd) are TSH cells, and that the ventral ones are GTH cells. Small GTH cells first appear in some fish at D(ay) 50 following fertilization (= D17 from hatching), when gonads are still undifferentiated. GTH cells increase greatly in number and size after D100. Contrary to treatment with 11β(OH)androstenedione, addition of methyl testosterone, progesterone and 17α(OH)progesterone stimulate the development of granular GTH cells in rainbow trout, shortly after hatching. It is suggested that steroid receptors in the brain or pituitary discriminate between 11-steroids and other steroids rather than between aromatizable androgens and other steroids.
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  • 82
    ISSN: 1432-0878
    Keywords: Vertebrate-type neuropeptides ; Neuroendocrine system of insects ; Neurosecretion ; Peptidergic neurons ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence and differential distribution of substances antigenically related to known vertebrate neuropeptides demonstrated within the corpus cardiacum of the insect Leucophaea are as follows: Of ten mammalian antisera tested, six yielded substantial immunoreactive deposits resembling oxytocin, somatostatin, Substance P, met-enkephalin, bombesin, and neurotensin, respectively. In the remaining four, the reaction was moderate (vasopressin, β-endorphin) or marginal (LH-RF, calcitonin). With regard to their regional distribution, these biochemically distinct reaction products seem to fall into two groups: (1) Materials resembling oxytocin, vasopressin, met-enkephalin, β-endorphin (and presumably also neurotensin and LH-RF) predominate in the central release area of the organ and are considered to be of extrinsic (cerebral) origin. (2) Substances localized primarily in areas rich in intrinsic glandular cells of the corpus cardiacum, and revealed by antisera raised against somatostatin, Substance P, and bombesin, are judged to be synthesized and stored within this organ. In peptidergic fibers entering the adjacent corpora allata, thus far Substance P-, β-endorphin-, and LH-RF-like immunoreactivities have been demonstrated. Some of these “new” neuropeptides may be contained in classical neurosecretory neurons, formerly identified by less specific methods, others must be assigned to additional peptidergic neurons heretofore unknown.
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  • 83
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Cardioactive peptide ; Lymnaea stagnalis ; Neurohormone ; Neurotransmitter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With an antiserum to the molluscan cardioactive tetrapeptide FMRF-amide immunoreactive perikarya and nerve fibers were identified in the central and peripheral nervous system of the pond snail Lymnaea stagnalis. Their localization is described. The same antiserum yielded reactive product in particular cells of the epithelium of the alimentary tract. The use of two different fixatives, glutaraldehyde, and a mixture of glutaraldehyde, picric acid, and acetic acid (GPA) showed that certain nerve cells can be identified only in material fixed with either the one or the other of these two fixatives, a result which indicates that in Lymnaea more than one FMRF-amide-like substance may occur. “Positive” axon endings were found in the periphery of various nerves, i.e., in places where neurohormones are released into the blood. Other fibers were found to end, probably synaptically, on other neurons, on epithelial cells in the stomach, and between muscle cells in various parts of the body, e.g., in the heart. In these cases the FMRF-amide-like substance may function as a neurotransmitter or a neuromodulator.
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  • 84
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    Keywords: Pancreatic polypeptide (PP)-like material ; Immunocytochemistry ; Nervous system ; Periplaneta americana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The central and visceral nervous systems of the cockroach Periplaneta americana were studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibody to bovine pancreatic polypeptide (PP). PP-like immunoreactive neuron somata are most numerous in the brain; at least 6 pairs of cell groups occur in clearly defined regions. Three pairs of cells each are also present in the suboesophageal ganglion and the thoracic ganglia, one pair of a single cell each in the first abdominal and the frontal ganglia, and 4 to 6 pairs of single cells in the terminal ganglion. No reactive cells were found in the retrocerebral complex and the second to the fifth abdominal ganglia. The axons containing PP-like immunoreactivity issue many branches that are distributed in the entire brain-retrocerebral complex, ventral cord, and visceral nervous system. PP-like immunoreactive material produced in the brain seems to be transported by three routes: protocerebrum to corpora cardiaca (-allata) through the nervi corporis cardiaci, tritocerebrum to visceral nervous system through frontal commissures, and to ventral cord through circumoesophageal connectives. A possible homology between the mammalian brain-GEP (gastro-enteropancreatic) system and the brain-midgut system of this insect is discussed.
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  • 85
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    Keywords: Luteinizing hormone ; releasing hormone ; Immunocytochemistry ; Distribution ; Forebrain ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of luteinizing hormone-releasing hormone (LHRH)-immunoreactive perikarya and processes was examined, in the untreated rat, with the unlabeled antibody enzyme method of immunocytochemistry on thick 50 μm vibratome sections. LHRH neurons were primarily observed in the preoptico-anterior hypothalamic and septal areas. Projections from these cell bodies to the median eminence form three distinct pathways, one laterally along the course of the optic tracts, one medially through the periventricular stratum of the third ventricle, and one through the tractus infundibularis. In addition, some of these cell bodies project to the organum vasculosum of the lamina terminalis (OVLT) and the subfornical organ (SFO). LHRH immunoreactive neurons were also noted in the anterior olfactory regions; they project along the medial olfactory tract to the olfactory bulb.
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  • 86
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    Keywords: Neuropeptides ; Substance P ; Uvea ; Immunocytochemistry ; Iris ; Smooth muscle contraction ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Substance P-immunoreactive nerve terminals were found in several locations in the anterior segment of the rabbit eye. In the iris they occurred in the sphincter muscle and were randomly distributed in the iris stroma with some fibres running close to the dilator muscle. In the ciliary body these immunoreactive elements were few and occurred within bundles of nerve fibres, while in the ciliary processes they were more numerous with a predominantly subepithelial location. Blood vessels in the anterior uvea were often surrounded by substance P-immunoreactive fibres. No substance P-fibres were found in the cornea, while the sclera contained very few such elements. Using conventional in vitro techniques it was found that the sphincter pupillae muscle of the iris responded to electrical stimulation with a contraction that was resistant to cholinergic and adrenergic blockade, but was inhibited by the neuronal blocker tetrodotoxin. This indicates the existence of a non-cholinergic, non-adrenergic neuronal mediator of the contractile response. Exogenously applied substance P produced a long-lasting contraction of the spincter muscle, an observation compatible with the view that substance P is the noncholinergic, non-adrenergic neurotransmitter involved.
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  • 87
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    Keywords: Cholecystokinin ; Spinal cord ; Immunocytochemistry ; Rat ; Neuropeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cholecystokinin-like immunoreactivity has been demonstrated by radioimmunoassay and immunocytochemistry in the spinal cord of various mammals, in particular in nerve fibers of the superficial layers of the posterior column, but had not been detected in neuronal cell bodies. We report immunohistochemical evidence for the presence of a group of cholecystokinincontaining neuronal cell bodies in the lumbar spinal cord of the rat. This group of cells is only visualized after direct injection of colchicine into the spinal cord and is located near the central canal in the intermedio-medial nucleus of area X of Rexed.
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  • 88
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    Keywords: Substance P ; Immunocytochemistry ; Neurohypophysis ; Hypothalamus ; Mouse
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Substance-P immunoreactivity has been located in semithin sections of mouse hypothalami and pituitaries and compared with the distribution of other hypothalamic peptides. In the mouse, nerve fibres and terminals reacting with antibodies against substance P (SP) were detected both in the external zone of the median eminence (ME) and in the neural lobe of the pituitary. Immunoreactive SP (ISP) axons of the ME did not react with antibodies against other peptides, i.e. arginine-vasopressin (AVP), oxytocin (OT), somatostatin and enkephalin, and were also negative with an antibody to serotonin. In the neural lobe, SP immunostaining occurred in AVP but not in OT axons. In the hypothalamus, ISP axons were widespread but conspicuously lacking in areas containing AVP neurones, i. e. in the suprachiasmatic nucleus and the clusters of AVP cells in the SO and PV nuclei. In contrast, multiple ISP endings were seen in contact with OT neurones. Immunoreactive cell bodies, only detected after colchicine treatment, belonged to two distinct classes of neurones: 1) single AVP neurones of the SO and PV nuclei; 2) specific (staining only for SP) neurones, scattered or grouped in different areas of the hypothalamus, not showing relationship with any circumscribed nucleus. These results reinforce the opinion that SP can be released as a neurohormone into the vascular portal system and can directly affect the pars distalis. The presence of immunoreactive SP in the neural lobe, which has not been reported in species other than the mouse, may have a different physiological significance.
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  • 89
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    Keywords: Substantia nigra (rat) ; Substance P ; Striatonigral pathway ; Immunocytochemistry ; Synaptic vesicles ; Synapses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A monoclonal antibody that recognises the C-terminal part of substance P was used to study immunoreactive structures in the substantia nigra by the unlabeled antibody, peroxidase-antiperoxidase procedure. Immunoreactivity was present in nerve fibres in all parts of the substantia nigra, particularly in the pars reticulata and pars lateralis. Electron microscopically two types of bouton immunoreactive for substance P were found: Type 1 contained large electron-lucent vesicles, occasional large granulated vesicles and formed symmetrical synapses with dendrites. Type 2 boutons contained smaller, round electron-lucent vesicles, many large granular vesicles and formed asymmetrical synapses (having prominent postjunctional dense bodies) with dendrites and perikarya. Immunoreactive fibres with varicosities that had been identified light microscopically were studied in serial sections in the electron microscope. Each identified varicosity contained synaptic vesicles and formed a single synapse. An individual fibre formed boutons of only one kind (type 1 or type 2) and could form multiple synapses with the same neuron. Thus, an identified fibre in the pars compacta had eight varicosities, each of which was in synaptic contacts (type 2) with the dendrites or soma of the same neuron. The results are consistent with the concept that substance P is a synaptic transmitter in the substantia nigra and indicate that neurons in this region may receive a significant input from substance P-containing afferents, and that there are at least two types of such afferent fibres.
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  • 90
    ISSN: 1432-0878
    Keywords: Somatostatin ; Immunocytochemistry ; CSF-contacting neurons ; Hypothalamus ; Xenopus laevis, tadpoles ; Amphibia, Anura
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In untreated tadpoles of Xenopus laevis, stage 60 (Nieuwkoop and Faber 1956), somatostatin-immunoreactive perikarya and fiber networks are already discernible in the pallium and the septum, as well as in the anterior and posterior hypothalamus. In addition, somatostatin-immunoreactive cerebrospinal fluid (CSF)-contacting neurons were found in the periventricular gray of the most caudal division of the ventral tuber cinereum. The results are discussed with respect to the inhibitory influence of the system of CSF-contacting and other somatostatin-immunoreactive neurons on the anterior pituitary and other target sites, especially during the climax of metamorphosis.
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  • 91
    ISSN: 1432-0878
    Keywords: Domestic fowl ; Hypothalamus ; LHRH-neurosecretion ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of LHRH-containing neuronal elements was investigated in the domestic fowl by means of immunohistochemical techniques. LHRH antisera were raised against synthetic LHRH in the rabbit. The antiserum used in the present study cross-reacted with LHRH of mammalian and avian tissues. LHRH-immunoreactive perikarya are located in the preoptic and in the septal areas, and in the bulbus olfactorius; however, no LHRH-immuno-reactive perikarya were found in the tuberal part of the hypothalamus. LHRH-immunoreactive fibers course from these areas toward the median eminence mainly along the wall of the third ventricle in the form of a periventricular network. Originating from the same cell groups other fibers run caudally immediately above the optic chiasma, forming the median bundle of the tractus preoptico-infundibularis. The third bundle running toward the OVLT is named the tractus preoptico-terminalis. In addition to these structures, LHRH-containing fibers and terminals were also present in different regions of the limbic system, in the dorsal part of the hippocampus, in the tuberculum and bulbus olfactorius, as well as in the optic lobe, nuclei commissurales tectales, organon subcommissurale, periaqueductal area, and pars ventralis mesencephali. The general distribution of the LHRH system in the chicken corresponds principally to that described previously in rodents (Sétáló et al. 1976, 1978). However, some subtle differences were demonstrated between the location of the LHRH system in birds and mammals.
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  • 92
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasoactive intestinal polypeptide ; Brain-gut peptide ; Pyloric antrum ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactivity to VIP by endocrine-like cells in the feline pyloric mucosa was examined by using three kinds of region-specific anti-porcine VIP sera. VIP-immunoreactive endocrine-like cells were detected clearly with all of the VIP antisera used. They were located mainly around the neck of the pyloric glands. Some of these endocrine-like cells showed dilution-dependent immunoreactivity against VIP antisera. The immunostaining intensity of VIP-immunoreactive endocrine-like cells showing dilution-independence could not be distinguished from those of nerve elements. The present results suggest that the immunoreactivity with properties very similar to those of authentic VIP may be present in the endocrine-like cells of the feline pyloric glands.
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  • 93
    ISSN: 1432-0878
    Keywords: Rat brain ; Hypothalamus ; Prolactin ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of prolactin (PRL)-like material is demonstrated in the brain of rats with the aid of anti-ovine PRL (oPRL) IgG as primary antibody in the unlabeled antibody-enzyme method. Immunoreactive deposits are visualized as an intraneuronal constituent with a widespread distribution in the hypothalamus and neural lobe of the pituitary. Dense networks of reactive nerve terminals derived from two prominent fibre tracts, a ventral (VHT) and a dorsal hypothalamo-neurohypophysial tract (DHT) are seen. The VHT is confined to the median eminence and pars oralis tuberis, the DHT to the pars caudalis tuberis. Both fibre tracts pass through the infundibular stalk into the neural lobe. The origin of the immunoreactive nerve terminals can be elucidated only to some extent. The VHT gives off beaded fibres entering the ependymal and glandular layer of the median eminence. Immunoreactive perikarya are observed in the supraoptic nucleus, the paraventricular nucleus, the anterior hypothalamic nucleus, the anterior commissural nucleus, the preoptic nucleus and the interstitial nucleus of the stria terminalis. A few of the immunoreactive perikarya are observed in close connection with brain vessels and the ependymal cells of the third ventricle. The results indicate that the anti-oPRL has a unique region specificity implying that only a segment of the mammalian PRL molecule is present in these nuclei of the brain. Fragments of PRL may function as neuromodulators or neurotransmitters in the rat brain.
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  • 94
    ISSN: 1432-0878
    Keywords: Cryo-ultramicrotomy ; Immunocytochemistry ; Pituitary gland ; Gonadotropic cells ; Salmo gairdneri
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The most frequently occurring cell types in the pars distalis of the pituitary gland of the rainbow trout, (i) the lactotropic, (ii) the gonadotropic, and (iii) the somatotropic cells, were identified in cryosections. Their morphological characteristics were compared with those of Epon-embedded material. Cell location, cell form, position of the nucleus, arrangement of rough endoplasmic reticulum and sizes of secretory granules proved to be useful parameters for identification. The size distribution of secretory granules of corresponding cells in cryosections and Epon sections proved to be similar. Additionally, both the immunoferritin and the unlabeled antibody enzyme method were applied for the immunocytochemical labeling of gonadotropic hormone-producing cells in cryosections. Anti-salmon-GTH as well as anti-carp-GTH serum showed the presence of GTH in both the smaller and the larger granules of the classical GTH cells, but also produced a reaction in TSH cells. Labelling of TSH cells was absent when using anti-β-carp-GTH. Specificity of the reaction depended upon the degree of dilution of the anti-GTH serum. Results with dilutions of 1∶4,000 and 1∶8,000 in the unlabeled antibody enzyme method, and of 1∶8,000 up to 1∶32,000 in the immunoferritin technique were optimal. Acid phosphatase activity in the smaller granules was demonstrated by enzyme cytochemistry in Epon sections. The relationship of the presence of hormone in these granules is discussed. The high sensitivity of the immunocytochemical labeling procedure is discussed with respect to cryo-ultramicrotomy.
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  • 95
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    Cell & tissue research 227 (1982), S. 67-77 
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide ; Neurosecretory cells ; Neuropeptides ; Calliphora vomitoria ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of a neuropeptide, previously shown to have the same or a very similar amino acid composition as vertebrate pancreatic polypeptide (PP), has been studied in the nervous system and gut of the blowfly, Calliphora vomitoria. Neurones immunoreactive to a bovine PP antiserum occur in the thoracic and abdominal ganglionic components of the central nervous system, in addition to the brain and suboesophageal ganglion. Pancreatic polypeptide appears to be relayed from its cells of origin to a neurohaemal organ in the dorsal sheath of the thoracic ganglion. PP immunoreactivity is also found in cells of the hypocerebral ganglion of the stomatogastric nervous system and in associated nerve fibres. The mid-gut contains PP-positive material in flask-shaped cells of its epithelial lining.
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  • 96
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    Cell & tissue research 227 (1982), S. 93-112 
    ISSN: 1432-0878
    Keywords: Neurophysins ; Immunocytochemistry ; Age-dependent changes ; Hypophysectomy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothalamo-extrahypophyseal neurophysin pathways (HEH) and the three hypothalamic nuclei secreting neurophysins, the supraoptic (SON), paraventricular (PVN) and suprachiasmatic (SCN) nuclei, of normal and hypophysectomized rats were studied by application of the immunoperoxidase procedure. Eight well-defined HEH pathways were recognized. Their main sites of projection were: lateral septum and subfornical organ (1 and 2); tractus diagonalis (3); medial nucleus of the amygdala and lateral ventricle (4); nucleus periventricularis thalami, nucleus habenulae lateralis and periaqueductal gray (5); periaqueductal gray, pineal organ, collicular recess and subependymal region of the fourth ventricle (6); dorsomedial nucleus and premammillary area (7); perimammillary region, corpus trapezoideum, ventral surface of medulla oblongata, nucleus tractus solitarii, nucleus commissuralis, substantia gelatinosa and formatio reticularis lateralis of the medulla oblongata and spinal cord (8). Neurophysin fibers of unknown origin were found in the frontal cerebral cortex. It was noted that in pathway 5 the amount of immunostainable material undergoes changes with age. The three neurophysin-secreting nuclei reacted differently following hypophysectomy. Among the HEH pathways the only one that seemed to be affected by hypophysectomy was that innervating the lateral septum. It is suggested that the neurons that survive hypophysectomy either do not project to the neural lobe or, alternatively, display axon collaterals projecting outside the neural lobe. Such a neuronal population could be the origin of the HEH pathways.
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  • 97
    ISSN: 1432-0878
    Keywords: Enolase (neuron-specific) ; S-100 protein ; Carotid body ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light- and electron-immunocytochemical investigation with the peroxidase-antiperoxidase (PAP) procedure revealed neuron-specific enolase and S-100 protein-like immunoreactivities specifically localized in the chief cells and the sustentacular cells of the rat carotid body, respectively. This finding suggests a neuron-like nature of the chief cells and a glia-like nature of the sustentacular cells on both embryological and functional bases.
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  • 98
    ISSN: 1432-0878
    Keywords: Newborn rat epidermis ; Soluble epidermal protein ; Thiolproteinase inhibitor ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Subcellular distribution of a thiol-proteinase inhibitor protein was determined in the epidermis of the newborn rat by light and electron microscopy. This protein was highly soluble in basal cells and concentrated on ribosomes in the perinuclear region. Solubility in Tris buffer decreased in granular and cornified cells in which the protein appeared on polysomes which were attached on other cellular structures such as dense homogenous deposits and tonofilaments. The protein also appeared to be deposited on the plasma membrane and became insoluble in Tris buffer at 37° C, but solubilized in 1 M phosphate buffer. Location of the protein around keratohyalin granules or by the plasma membrane suggested that the inhibitor protein bound to cysteinerich protein of the epidermis with or without forming a thiol-proteinase inhibitor complex. The thiol-proteinase inhibitor protein seems to contribute to epidermal cell differentiation at multiple points through changes in its solubility and subcellular localization from basal cells to cornified cells.
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 223 (1982), S. 627-639 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Progesterone-binding protein (PBP) ; Placenta ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular localization of progesterone-binding protein (PBP) in the guinea-pig placenta was studied by use of immunocytochemical procedures. Within the chorioallantoic placenta, a strong positive reaction was observed in the interlobar and marginal trophoblast from the third week of gestation to term. PBP was localized in the cytoplasm of the syncytiotrophoblast, and the nuclei were never stained. At the ultrastructural level, the immunoreaction was associated with the rough endoplasmic reticulum, the Golgi apparatus and the perinuclear space. No deposits were seen in any other cell organelles. This localization strongly suggests that the interlobar syncytium is related to PBP synthesis. In the labyrinth, a weak immunoreaction was observed by light microscopy around some blood lacunae. At the ultrastructural level the dense deposits were localized in vesicles located near the maternal lacunae. The distribution of PBP was also studied by light microscopy in other tissues from pregnant guinea-pig. No PBP, or PBP-like material, was detected inside cells from liver, muscle, heart, lung, kidney, ovary, and uterus. A weak immunoreaction for PBP was detected in vascularized zones of these organs. These observations strongly suggest that PBP, a protein related to gestation in the guinea-pig, is elaborated by the placental tissue of this hystricomorph rodent. PBP is the first steroid-binding plasma protein shown to be of extrahepatic origin.
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  • 100
    ISSN: 1432-0878
    Keywords: Gonadotrophs ; Male rats ; Immunocytochemistry ; Cytophotometry ; Image analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Quantitative studies on the population of the gonadotropic cells in the pituitary of adult male rats were performed after trypsic dissociation of the pituitary glands and immunoenzymatic staining with anti-β-LH or anti-β-FSH antisera. Number, area and extinction of labelled cells were measured by use of an image analyser and a cytophotometer. The gonadotrophs represent approximately 14% of the pituitary cells. The mean area of gonadotrophs is significantly larger after staining with anti-β-LH serum than after staining with anti-β-FSH serum. Planimetric measurement of the gonadotrophs reveals their variability in size ranging between 30 and 160 μm2. Moreover, the size distribution depends on the staining serum used: more numerous small-sized cells (〈75 μm2) are stained with anti-β-FSH serum than with anti-β-LH serum, which conversely stains more numerous large-sized cells. Cytophotometric measurements indicate that immunostaining varies greatly among cells of the same size class and that the staining intensity appears to increase according to the cell size. These results emphasize the morphofunctional heterogeneity of the gonadotropic cell population.
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