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  • Immunocytochemistry  (149)
  • Springer  (149)
  • 1980-1984  (149)
  • 1925-1929
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  • 1
    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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  • 2
    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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  • 3
    ISSN: 1432-2048
    Keywords: Cotyledons ; Endoplasmic reticulum ; Ferritin labeling ; Immunocytochemistry ; Phaseolus ; Protein (reserve) ; Reserve protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ultrastructure of the storage parenchyma cells of the cotyledons of developing bean (Phaseolus vulgaris L.) seeds was examined in ultrathin frozen sections of specimens fixed in a mixture of glutaraldehyde, formaldehyde and acrolein, infused with 1 M sucrose, and sectioned at-80° C. Ultrastructural preservation was excellent and the various subcellular organelles could readily be identified in sections which had been stained with uranyl acetate and embedded in Carbowax and methylcellulose. The cells contained large protein bodies, numerous long endoplasmic reticulum cisternae, mitochondria, dictyosomes, and electron-dense vesicles ranging in size from 0.2 to 1.0 μm. Indirect immunolabelling using rabbit immunoglobulin G against purified phaseolin (7S reserve protein), and ferritin-conjugated goat immunoglobulin G against rabbit immunoglobulin G was used to localize phaseolin. With a concentration of 0.1 mg/ml of anti-phaseolin immunoglobin G, heavy labeling with ferritin particles was observed ober the protein bodies, the cisternae of the endoplasmic reticulum, and the vesicles. The same structures were lightly labeled when the concentration of the primary antigen was 0.02 mg/ml. Ferritin particles were also found over the Golgi bodies. The absence of ferritin particles from other organelles such as mitochondria and from areas of cytoplasm devoid of organelles indicated the specificity of the staining, especially at the lower concentration of anti-phaseolin immunoglobulin G.
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  • 4
    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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  • 5
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; (PEP carboxylase) ; PEP carboxylase ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The localization of phosphoenol pyruvate carboxylase (EC 4.1.1.3.1.) in the leaf cells of Sorghum vulgare was investigated by using three techniques: the conventional aqueous and non aqueous methods gave conflicting results; the immunocytochemical techniques clearly showed that the enzyme is predominantly located in the cytoplasm of mesophyll cells.
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  • 6
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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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  • 7
    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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  • 8
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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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  • 9
    Electronic Resource
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    Protoplasma 105 (1981), S. 333-339 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Pea seed ; Protein histochemistry ; Storage proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The pea seed storage proteins legumin and vicilin have been localized by electron microscopy using a post-embedding immunocytochemical double-labelling technique. Highly purified antibodies from sheep, specific to legumin and vicilin, were sequentially reacted on sections of pea cotyledon tissue embedded in glycol methacrylate or Spurr's epoxy resin. The sheep antibodies were visualized indirectly by reaction with Staphylococcal Protein A labelled with colloidal gold. Two discrete sizes of colloidal gold particles can be used for simultaneous localization of two antigens. In near mature tissue, the storage proteins are restricted to organelles, called protein bodies. Individual protein bodies contain both legumin and vicilin.
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  • 10
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    Cell & tissue research 208 (1980), S. 499-505 
    ISSN: 1432-0878
    Keywords: Somatostatin-immunoreactive elements ; Brain ; Lizard ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The brain of the lizard, Ctenosauria pectinata, was studied light microscopically using an immunocytochemical staining method that is specific for neurohypophysial hormones and somatostatin. It was shown that the telencephalon and particularly the diencephalon contain somatostatin-producing perikarya, while somatostatinergic fibers occur in the entire brain. Similar to the situation in other vertebrates, somatostatin neurons in Ctenosauria pectinata form a population distinct from the neurohypophysial hormone-producing neurons. The small-sized somatostatin neurons were found in the cortex and the hypopallium of the telencephalon, and in two distinct clusters in the diencephalon: (1) ventral from, and partially overlapping with, the classical neurosecretory para ventricular nucleus; and (2) in the region of the infundibular (tuberal) nucleus. Somatostatin fibers were found among the classical neurosecretory fibers of the supraoptico-paraventricular system (tract, median eminence, neural lobe), near to and within the epiphysis, in the septum, in the vicinity of the tectum opticum and the cerebellum, and in the tegmentum.
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  • 11
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    Cell & tissue research 209 (1980), S. 365-370 
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide (PP) cells ; Paracrine secretion ; Human pancreas ; Immunocytochemistry ; Pancreatic endocrine tumours
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pancreatic polypeptide (PP) cells were studied in human endocrine pancreatic tumours and in normal human pancreata by immunohistochemical techniques and electron microscopy. The existence of long cytoplasmic processes was demonstrated both in tumours and normal tissue. These processes are in close contact with other endocrine cells or with acinar cells. This particular morphological aspect suggests that PP cells may control the function of other cells via paracrine secretion.
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  • 12
    ISSN: 1432-0878
    Keywords: Pancreatic polypeptide (PP)-like material ; Immunocytochemistry ; Neuropeptides ; Neurosecretory cells ; Calliphora erythrocephala
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The brain of the blowfly, Calliphora erythrocephala, has been studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibodies to bovine pancreatic polypeptide (BPP). A number of immunoreactive neurones have been localised, some corresponding to neurones previously identified tentatively as neurosecretory. This finding is further evidence that biologically active peptides, previously considered to be “vertebrate”, also exist in invertebrates. It also supports the concept of their evolutionary origin in nervous tissue.
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  • 13
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    Cell & tissue research 215 (1981), S. 499-504 
    ISSN: 1432-0878
    Keywords: Androgen receptors ; Pituitary gland ; Rhesus monkey ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The uptake and retention of radiolabelled dihydrotestosterone by the pituitary gland was examined in the rhesus monkey. Two animals were given an intravenous injection of 1.0μg/kg 3H-dihydrotestosterone (DHT) alone while one monkey received both the labelled androgen and 100μg/kg of unlabelled steroid. One and a half hours later, they were sacrificed. The pituitary glands were removed and processed for autoradiography and immunocytochemistry. Autoradiographic localization of DHT was discernible in the partes nervosa, intermedia and distalis, albeit the highest concentration of radiolabelled cells was noted in the pars distalis. Immunocytochemical staining with antibodies to rat PRL, human TSHβ and ovine LHβ revealed a population of steroid-concentrating cells that contained TSH and a second group that contained LH. None of the cells that reacted with the anti-PRL serum were radiolabelled.
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  • 14
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    Cell & tissue research 215 (1981), S. 75-86 
    ISSN: 1432-0878
    Keywords: Pituitary cells ; Estrogen ; Progesterone ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of progesterone pretreatment on in vivo 3H-estrogen uptake by five anterior pituitary cell types was analyzed by means of a quantitative autoradiographic-immunocytochemical technique. Male and female rats castrated for 14 days show nuclear concentration of label in all five cell types one h after injection of 3H-estradiol, whether progesterone treated or not. The order of labeling intensity is gonadotropes ≧ lactotropes = somatotropes 〉 thyrotropes = corticotropes. Progesterone treatment induces a dramatic sexual dimorphism in estrogen uptake; it significantly increases 3H-estrogen uptake in all female cell types. In males, progesterone decreases uptake in gonadotropes while not altering uptake in other cell types.
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  • 15
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biologically active peptides ; Lymnaea stagnalis ; Neurohormone ; Neurotransmitter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Perikarya and nerve fibers were immunocytochemically identified in the central nervous system of the pond snail Lymnaea stagnalis by means of the unlabelled antibody enzyme method with antisera to 15 biologically active peptides of vertebrates: vasopressin, vasotocin, oxytocin, α-melanocyte stimulating hormone (α-MSH), met-enkephalin, somatostatin, glucagon, insulin, glucose-dependent insulinotropic peptide (GIP), vaso-active intestinal polypeptide (VIP), gastrin, secretin, pancreatic polypeptide (PP), Substance P, calcitonin. No immunostaining was obtained with antisera to β-endorphin, cholecystokinin (CCK), neurophysin I and II. Particular neurons could be identified with two antisera (anti-vasopressin/vasotocin, anti-α-MSH/metenkephalin, anti-substance P/PP, anti-PP/gastrin). Apparently this indicates that populations of cells identified with a given antiserum may consist of more than one cell type. Only a few of the new peptidergic cells appeared to be identical with classical neurosecretory cells. Thus the growth hormone producing Light Green Cells stained with anti-somatostatin and the axon terminals of the ovulation hormone producing Caudo-Dorsal Cells with anti-met-enkephalin. Whether this indicates structural identity of the growth hormone with somatostatin and of the ovulation hormone with met-enkephalin remains to be investigated. Just like the classical neurosecretory cells a number of the new peptidergic cells (anti-glucagon, -insulin, -met-enkephalin, -somatostatin, and -PP positive cells) send their axons to the peripheries of commissures, connectives or nerves. Thus these cells can be considered as probably neuroendocrine. The classical neurosecretory cells release their products into the haemolymph from these sites. Other new peptidergic cells (e.g., anti-vasopressin, -vasotocin, -oxytocin and -GIP positive cells) have axons that terminate, probably synaptically, on other neurons, indicating that they are “more conventional” neurons, their products being neurotransmitters/neuromodulators. It can also not be excluded that some cells of a population containing a given peptide are neuroendocrine and others make contact with other neurons.
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  • 16
    ISSN: 1432-0878
    Keywords: Pituitary ; Immunocytochemistry ; Prolactin ; Somatotropin ; Lepidosiren
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular binding sites of anti-oPRL IgG and anti-bSTH IgG were demonstrated in the pituitary glands of Lepidosiren paradoxa, Rana temporaria and Ambystoma mexicanum by means of the unlabeled antibody enzyme method by light and electron microscopy (the latter only in Lepidosiren). With the light microscope PRL or PRL-like substances and STH or STH-like substances were revealed in two different cell types in the distal lobe corresponding to the acidophils. However, as a result of the insufficient differentiation of the acidophils in Lepidosiren after staining with Brookes' procedure it was not possible to distinguish the two types of acidophils in this animal. Treatment with low dilutions of both anti-oPRL and anti-bSTH IgG revealed simultaneous immunocytochemical staining in both types of acidophils in Lepidosiren and in Rana. These results, indicating that there is antigenic cross-reaction between anti-oPRL and anti-bSTH IgG and both PRL and STH in these animals, are discussed. The electron microscopic investigations of Lepidosiren revealed that the specific anti-oPRL IgG reactive cells contain granules ranging in size from 200 to 300 nm, while the specific anti-bSTH IgG reactive cells contain smaller immunoreactive granules ranging from 80 to 160nm.
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  • 17
    ISSN: 1432-0878
    Keywords: Hypothalamus (rat) ; Differentiation ; Transplant ; Histofluorescence ; Immunocytochemistry ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Hypothalamic tissue from 16 to 18-day fetal rats was transplanted onto the choroidal pia overlying the superior colliculus in adult female rats. After survival periods of 2 weeks to 19 months, brains containing transplants were processed for monoamine fluorescence histochemistry, immunohistochemistry for three neuropeptides (LHRH, somatostatin, neurophysin), or for autoradiography in ovariectomized hosts that received [3H] estradiol. Most of the transplants survived and retained or increased in size; 14 of 25 transplants examined by fluorescence histochemistry were found to contain median eminence-like structures. In almost all of the transplants that were stained for neuropeptides, beaded processes and occasional cell bodies were observed. Although immunoreactive fibers were found near blood vessels, no palisade arrangement typical of the normal median eminence was evident. Each of the hypothalamic transplants on which steroid autoradiography was performed contained clusters of estrophilic neurons, the intensity of labeling of which was comparable to that seen in the host hypothalamus. These results indicate that many characteristic morphological and chemical features of the hypothalamus, which are not evident in the 16 to 18-day fetus, are elaborated in transplants during the survival period in the host. Transplantation of fetal hypothalamus to adult choroidal pia thus appears to be a valuable approach for studying the factors, humoral or neural, that regulate the differentiation of this brain region.
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  • 18
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    Cell & tissue research 206 (1980), S. 355-365 
    ISSN: 1432-0878
    Keywords: LRH neurons ; Prosimians (Tupaia, Galago) ; Fluorescence ; microscopy ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Reactive LRH neurons were characterized in prosimians (Tupaia and Galago) by immunofluorescence using rabbit immunesera against unconjugated synthetic LRH, or LRH conjugated with bovine serum albumin. These neurons, which vary individually in number in one species, are mainly concentrated in the rostral hypothalamus (medial preoptic area and anterior hypothalamic area) and in the lamina terminalis. In contrast to the simians and man, immunoreactive perikarya were not routinely found in the mediobasal hypothalamus of the prosimians investigated in the present study. Reactive axons of the hypothalamo-hypophyseal tract are more numerous and conspicuous in the retrochiasmatic area and in the postinfundibular eminence. They give rise to radiating collaterals ending mainly around the capillaries of the primary portal plexus of the median eminence and of the infundibular stem (where they are generally more numerous). Reactive axons of the preopticoterminal tract, originating from the perikarya of the lamina terminalis, end around the capillaries of the vascular organ or below and between the ependymal cells lining its ventricular side. In Galago a small but very distinct tract of reactive axons runs under the optic chiasma, between the lamina terminalis and the ventral labium of the infundibulum. Very fine reactive extrahypothalamic axons were observed in the posterior part of the habenular ganglia, along the preamygdaloid portion of the stria terminalis and along the blood vessels of the parolfactory area.
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  • 19
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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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  • 20
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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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  • 21
    ISSN: 1432-0878
    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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  • 22
    ISSN: 1432-0878
    Keywords: Neurotensin ; Hypothalamus ; Coturnix quail ; Immunocytochemistry ; Neuropeptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hypothalamus of Japanese quail, Coturnix coturnix japonica, has been studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibodies to synthetic neurotensin (NT). A number of immunoreactive neuronal perikarya occur in the medial preoptic nucleus of the rostral hypothalamus and a few in the accessory part of paraventricular nucleus and dorsal portion of the infundibular nucleus. Some of them correspond to the parvocellular neurons previously identified tentatively as neurosecretory (Mikami et al. 1975, 1976). Large numbers of immunoreactive neuronal fibers are found in the preoptic area, which extend as a remarkable fiber tract from this area to the ventral septal area and to the subfornical organ. A few immunoreactive fibers also extend ventrocaudally to the infundibular nucleus and to the neural lobe.
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  • 23
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    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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  • 24
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    Cell & tissue research 225 (1982), S. 79-93 
    ISSN: 1432-0878
    Keywords: Immunoreactive prolactin ; Immunocytochemistry ; Circumventricular organs ; Choroid plexus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunoreactive prolactin (IMP) has been localized in the male rat brain using the soluble peroxidase-anti-peroxidase (PAP) technique. In normal untreated animals, reaction product was seen in choroid plexus (CP) and in ependymal cells of the ventricular lining with heaviest concentrations of positively staining cells in the 3rd ventricle near the subcommisural organ (SCO), in the lateral ventricles near the subfornical organ (SFO), and in the 4th ventricle near the area postrema (AP). IMP was also present in numerous ependymal cells resembling tanycytes in the cerebral aqueduct, central canal of the spinal cord at the level of the AP, the organum vasculosum of the lamina terminalis (OVLT) and the floor of the infundibular recess. Immunoreactive cells resembling neurons were localized within the substance of the AP, SCO, and OVLT. IMP was also present in fibers of the zona externa of the median eminence and infundibular stalk; a few cells of the pars tuberalis contained reaction product. Hypophysectomized rats and bromocriptine-treated rats exhibited a similar staining pattern except that bromocriptine treatment eliminated IMP from most CP cells. Hypophysectomy, bromocriptine or estrogen treatment enhanced staining for IMP in cells of the pars tuberalis; estrogen treatment or hypophysectomy produced an increase in the number and distribution of immunoreactive cells as well as increased density of reaction product in cells of the medial habenular nucleus. The functional relevance of prolactin in these locations in the brain, the possible routes of transport of prolactin from the pituitary gland to the central nervous system, and the strong suggestion of extra-pituitary sites of synthesis of a prolactin-like hormone are discussed.
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  • 25
    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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  • 26
    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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  • 27
    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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  • 28
    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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  • 29
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    Cell & tissue research 226 (1982), S. 231-235 
    ISSN: 1432-0878
    Keywords: Secretin-like cells ; Immunocytochemistry ; Digestive tract ; Styela plicata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Secretin-like cells have been detected in the digestive tract of the ascidian Styela plicata by means of immunofluorescent and immunocytochemical methods. Especially, in the esophageal epithelium there are immunoreactive cells (S2) in which a biogenic amine (5-HT) and a regulatory peptide (secretin) occur together. In the gastric epithelium only secretin-like cells (S1) are present. Tests of cross-reactivity performed with glucagon, GIF and VIP, have confirmed the presence of a secretin-like molecule only in the S1 and S2 cells.
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  • 30
    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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  • 31
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    Cell & tissue research 227 (1982), S. 423-427 
    ISSN: 1432-0878
    Keywords: Liver (Rat) ; Albumin ; Metabolism ; Hepatocytes ; Immunocytochemistry ; Nutrition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunoperoxidase method was used to identify albumin in hepatocytes of rats before and after periods of starvation. All hepatocytes in fed rats contained a relatively large amount of nascent albumin. Overnight fasting reduced the number of hepatocytes with a large amount of albumin to primarily those surrounding terminal hepatic venules. These were estimated to be about 30% of the population. The other cells had only a slight amount of albumin. After 48 h of fasting all hepatocytes contained a low level of albumin.
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  • 32
    ISSN: 1432-0878
    Keywords: Enolase (neuron-specific) ; S-100 protein ; Carotid body ; Immunocytochemistry
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    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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  • 33
    ISSN: 1432-0878
    Keywords: Nucleus lateralis tuberis ; Hypothalamo-hypophysial relationship ; Immunocytochemistry ; Prolactin cells ; Teleosts
    Source: Springer Online Journal Archives 1860-2000
    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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  • 34
    ISSN: 1432-0878
    Keywords: Villin ; Intestinal brush border ; Contractile proteins ; Immunocytochemistry ; Immunostaining of blotted gels
    Source: Springer Online Journal Archives 1860-2000
    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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  • 35
    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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  • 36
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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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  • 37
    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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  • 38
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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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  • 39
    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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  • 40
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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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  • 41
    ISSN: 1432-0878
    Keywords: Somatotrops ; Oncorhynchus ; Immunocytochemistry ; Immunodiffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pituitary somatotrops were localized in four species from the genus Oncorhynchus using an antiserum to chum salmon (Oncorhynchus keta) growth hormone. The antiserum cross-reacted specifically with the acidophils in the caudal pars distalis of all species tested. Corroborative immunodiffusion studies were also conducted with pituitary homogenates from the same species. Immunological identity of the GH molecule was observed for all species tested.
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  • 42
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    Cell & tissue research 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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  • 43
    ISSN: 1432-0878
    Keywords: ACTH-opiocortin system ; Immunocytochemistry ; Median eminence ; Neural lobe
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary By immunocytochemical methods, the present study describes ACTH-immunoreactive fibers in the pituitary stalk and neural lobe. This opiocortin-hypothalamo-neural lobe projection arises in a bed nucleus of perikarya in the basal hypothalamus, follows supraoptico-hypophyseal fibers in the zona interna of the median eminence, and distributes throughout the neural lobe. No ACTH-immunoreactive fibers project to the zona externa; some are present in the subependymal layer and at the lateral margins of the median eminence. Further studies must identify the role of these fibers in posterior lobe function. It remains also to be determined whether this system terminates upon primary pituitary portal capillaries and delivers opiocortin neuropeptides to the adenohypophysis.
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  • 44
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Human TSH β subunit ; Teleosts ; TSH cells
    Source: Springer Online Journal Archives 1860-2000
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    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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  • 45
    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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  • 46
    ISSN: 1432-0878
    Keywords: Proctolin ; Periplaneta americana ; Terminal ganglion-hindgut system ; Immunocytochemistry ; Cobalt iontophoresis technique
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the American cockroach, the distribution and connections of neuronal elements of the terminal ganglion-proctodeal nerve-hindgut system were investigated by means of immunohistochemical methods and axonal CoCl2 iontophoresis. Proctolinlike immunoreactivity was localized within neurons of the terminal ganglion projecting into the proctodeal nerve on the one hand, and in nerve cells without a direct connection to this system on the other. Immunohistochemically, in whole mount preparations fibres of the proctodeal nerve and terminal structures in the hindgut musculature exhibit strong proctolinlike immunoreactivity. At the light- and electron-microscopic levels the pathways of about 30 somata of the proctodeal neural system were characterized by cobalt chloride iontophoresis. The relationships of cobalt filled and immunoreactive neuronal structures are discussed. For the preparation of tritiated proctolin we thank Dr. S. Reißmann, WB Biochemie, Sektion Biologie, FSU Jena
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  • 47
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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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  • 48
    ISSN: 1432-0878
    Keywords: Retinal photoreceptors ; Opsin ; Optic nerve ; Immunocytochemistry ; Cyclostome, Myxine glutinosa
    Source: Springer Online Journal Archives 1860-2000
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    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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  • 49
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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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  • 50
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    Cell & tissue research 234 (1983), S. 125-134 
    ISSN: 1432-0878
    Keywords: Vasopressin ; Immunocytochemistry ; Electron microscopy ; Hypothalamus ; Rat
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    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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  • 51
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    Cell & tissue research 234 (1983), S. 319-333 
    ISSN: 1432-0878
    Keywords: Serotonin ; Goldfish ; Immunocytochemistry ; Central nervous system
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    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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  • 52
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    Keywords: Urotensin ; Caudal neurosecretory system ; Immunocytochemistry
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    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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  • 53
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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
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    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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  • 54
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    Keywords: FMRFamide ; Bovine pancreatic polypeptide ; Immunocytochemistry ; Peptidergic neurons ; Leptinotarsa decemlineata
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    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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  • 55
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    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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  • 56
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    Cell & tissue research 236 (1984), S. 171-180 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
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    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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  • 57
    ISSN: 1432-0878
    Keywords: ACTH cells ; Immunocytochemistry ; Morphometry ; Spontaneous hypertension
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    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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  • 58
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    Keywords: Corticotropin-releasing factor ; Immunocytochemistry ; Hypothalamus ; Domestic fowl
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    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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  • 59
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    Keywords: Pituitary gland ; Goat ; Folliculo-stellate cell ; GH cell ; S-100 protein ; Immunocytochemistry
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    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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  • 60
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    Cell & tissue research 212 (1980), S. 63-72 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Brush border ; Epithelial cells ; Vitamin D ; Cell maturation
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    Topics: Biology , Medicine
    Notes: Summary The vitamin D-dependent calcium-binding protein (CaBP) was studied in relation to the age of the cell, in isolated epithelial cell populations removed from rat duodenum. Alkaline phosphatase and thymidine kinase activities were used as markers to characterize differentiated villus cells and undifferentiated (mitotically active) crypt cells, respectively. CaBP distribution along the length of the villus, as established by radioimmunoassay, appears as a gradient increasing from the crypt to the tip of the villus. CaBP concentration in cells is shown to be (i) negatively correlated with the thymidine kinase activity of cells, and (ii) positively correlated with the alkaline phosphatase activity of cells. This indicates that CaBP is absent in crypt cells and appears in differentiated cells with the development of the brush border. Thus CaBP, like alkaline phosphatase, can be considered as an indicator of enterocyte maturation. These data were also confirmed by studying the cellular localization of the protein. In addition both indirect immunofluorescence and immunoperoxidase staining methods reveal that antibody against CaBP decorates the terminal web, but not the microvilli of the brush border of mature absorptive cells. The results suggest that CaBP may act as a modulator of some Ca2+-mediated biochemical processes at the level of the enterocyte brush border.
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  • 61
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    Keywords: α- and β-Endorphins ; Met-enkephalin ; Brain ; Immunocytochemistry ; Rana temporaria (Amphibia, Anura)
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    Notes: Summary In the brain of Rana temporaria, two distinct systems reactive with α- and β-endorphin antisera, respectively, and with a met-enkephalin antiserum, have been detected immunohistochemically. Neurons reacting with α- and β-endorphin antisera are located (1) in the preoptic nucleus, and (2) in the pars ventralis of the tuber cinereum. Immunoreactive nerve fibres of both groups of perikarya end in the infundibular floor near the capillaries and the preoptico-hypophysial tract. Control reactions have shown that the immunoreactivity is suppressed by the corresponding antigens, but also by β-LPH. In view of these results the immunoreactive systems examined correspond to an α/β-endorphin system or a lipotropinergic system. Neurons reacting with the met-enkephalin antiserum are located in the paraventricular organ. Intense immunofluorescence was observed in the infundibular floor. Controls show that the labelling by met-enkephalin antiserum is exclusively suppressed by met-enkephalin. In the pituitary gland, on the other hand, α- and β-endorphin antisera reveal: 1) the MSH/ACTH-like cells of the pars intermedia and 2) the ACTH-like cells of the pars distalis.
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  • 62
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    Keywords: Digestive tract ; Fish ; Gastrin-immunoreactive cells ; Pancreatic polypeptide-immunoreactive cells ; Somatostatin-immunoreactive cells ; Immunocytochemistry
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    Notes: Summary Gastrin, pancreatic polypeptide and somatostatin immunoreactive cells in the gut of two fish with stomachs (perch and catfish) and a stomachless fish (carp) were studied by immunocytochemistry. In the gastric mucosa of perch and catfish, cells showing gastrin and somatostatin-like immunoreactivity are found, scattered among the surface mucous cells and mucous neck cells. No pancreatic polypeptide (P.P.) immunoreactive cells are detected in the gastric mucosa. Cells showing gastrin and P.P.-like immunoreactivity are observed in the intestinal mucosa of perch, catfish and carp. In this location no somatostatin immunoreactive cells are found.
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  • 63
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    Cell & tissue research 221 (1981), S. 257-267 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Human pituitary hormone antisera ; Platyfish ; Pituitary gland
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    Topics: Biology , Medicine
    Notes: Summary In this communication we describe the immunocytochemical cross-reactivity between antisera to various human pituitary hormones and specific hormone producing cell types in the pituitary gland of sexually mature male platyfish (Xiphophorus maculatus). Antisera to human pituitary hormones cross-reacted either with cells known to produce corresponding hormones (or hormone subunits) in the platyfish (e.g., ACTH, prolactin, TSH β, LH α, FSH α, TSH α) or with no pituitary cells at all (e.g., LH β, FSH β). The one exception was antiserum to human growth hormone which cross-reacted with MSH and ACTH producing cells. The platyfish pituitary is proposed as a test system for immunocytochemically screening antisera for purity and specificity in order to determine their applicability in particular studies.
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  • 64
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    Cell & tissue research 220 (1981), S. 279-292 
    ISSN: 1432-0878
    Keywords: Endocrine cells ; Gastrointestinal tract ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary The distribution of gastrin-, cholecystokinin-, glucagon-, secretin-, vasoactive intestinal polypeptide-, substance P-, bombesin-, neurotensin-, motilin-, somatostatin- and avian pancreatic polypeptide-like cells, demonstrated by indirect immunocytochemistry, was studied in samples from the following regions: proventriculus, gizzard, pylorus, duodenum, upper and lower ileum, caeca and rectum. The pylorus is particularly rich in gastrin-, neurotensin- and somatostatin-like cells. No cells immunoreactive for gastric inhibitory polypeptide or insulin were detected. In a number of instances the same cells were found to stain with antisera raised to different gut peptides. This happened with antisera detecting gastrin- and neurotensin-like cells, with antisera to avian pancreatic polypeptide and glucagon and with antisera to secretin, vasoactive intestinal polypeptide, glucagon and substance P. The possibility that antigenic determinants to more than one peptide are contained in certain endocrine-like cells is considered.
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  • 65
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    Cell & tissue research 219 (1981), S. 213-216 
    ISSN: 1432-0878
    Keywords: Relaxin ; Human pregnancy ; Corpus luteum ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary Corpora lutea from 12 pregnant women were prepared for immunohistochemical localization of relaxin using a highly specific antiserum. A positive response is given by luteal cells that are diffusely distributed throughout the corpus luteum. These cells do not form a distinctive group in any particular area. A negative response is seen in the adjacent ovarian tissue, and also in nongestational corpora lutea in an early luteal phase.
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  • 66
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    Keywords: Hypothalamic deafferentation ; Immunocytochemistry ; Vasopressin ; ACTH/opiocortin ; LHRH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Luteinizing hormone-releasing hormone (LHRH), vasopressin, and corticotropin systems were examined by immunocytochemical methods in male rats 2 to 20 days after deafferentation of the basal hypothalamus. Axonal degeneration of the vasopressin system (whose perikarya lie rostral to the island) and the corticotropin system (whose perikarya lie within the island) was examined and compared with the response of the LHRH system. Vasopressin immunoreactive staining was absent in the internal zone of the median eminence 10 and 20 days after deafferentation. Disruption of the efferent projections of the opiocortin system caused the loss of almost all fiber staining outside the island by the 5th postoperative day. LHRH staining in the median eminence was modestly reduced in 5 days, considerably reduced in 10 days and negligible 20 days after deafferentation. At 10 and/or 20 days after deafferentation densely stained fibers of all three systems were observed on both sides of the cut. Invasive vasopressinergic fibers reached the lateral median eminence by the 20th postoperative day. This study reports on the response of three neuropeptide systems after complete deafferentation and demonstrates that regeneration can occur across the knife cut.
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  • 67
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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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  • 68
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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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  • 69
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    Cell & tissue research 227 (1982), S. 93-112 
    ISSN: 1432-0878
    Keywords: Neurophysins ; Immunocytochemistry ; Age-dependent changes ; Hypophysectomy ; Rat
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    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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  • 70
    ISSN: 1432-0878
    Keywords: Enteroendocrine cells ; Pancreatic endocrine cells ; Gastroenteropancreatic hormones ; Immunocytochemistry ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Four immunoreactive endocrine cell types can be distinguished in the pancreatic islets of B. conchonius: insulin-producing B cells, somatostatin-producing A1 (= D) cells, glucagon-producing A2 cells and pancreatic poly-peptide-producing PP cells. The principal islet of this species contains only a few PP cells, while many PP cells are present in the smaller islets. Except for the B cell all pancreatic endocrine cell types are also present in the pancreatic duct. At least six enteroendocrine cell types are present in the gut of B. conchonius: 1. a cell type (I) with small secretory granules, present throughout the intestine, and possibly involved in the regulation of gut motility; 2. a C-terminal gastrin immunoreactive cell, probably producing a caerulein-like peptide; these cells are located at the upper parts of the folds, especially in the proximal part of the intestinal bulb; 3. a met-enkephalin-immunoreactive cell, present throughout the first segment; 4. a glucagon-immunoreactive cell, which is rare in the first segment; 5. a PP-immunoreactive cell, mainly present in the first half of the first segment; 6. an immunoreactive cell, which cannot at present be specified, located in the intestinal bulb. The latter four cell types are mostly located in the basal parts of the folds, although some PP-immunoreactive cells can also be found in the upper parts. Most if not all enteroendocrine cells are of the open type. The possible functions of all enteroendocrine cell types are discussed.
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  • 71
    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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  • 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
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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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  • 74
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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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  • 75
    ISSN: 1432-0878
    Keywords: Crustacean hyperglycemic hormone ; Astacus leptodactylus ; Immunocytochemistry ; Quantitative electron microscopy ; Secretory cycle
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    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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  • 76
    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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  • 77
    ISSN: 1432-0878
    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; 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 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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  • 78
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    Cell & tissue research 215 (1981), S. 41-45 
    ISSN: 1432-0878
    Keywords: Hypothalamic lesions ; Somatostatin fibers ; Median eminence ; Neural lobe ; Immunocytochemistry ; Frog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A light microscopic immunocytochemical study of the brain of frogs with hypothalamic lesions was performed in order to obtain evidence concerning the origin of somatostatin fibers in the median eminence and neural lobe of the hypophysis. The results indicate that the somatostatin fibers of the neural lobe originate from somatostatin perikarya located in the prechiasmatic part of the hypothalamus and possibly also in the telencephalon. The somatostatin neurons of the pars ventralis tuberis cinerei do not send axons to the neural lobe. The frog median eminence contains axon terminals of somatostatin neurons located in the pars ventralis of the tuber cinereum. Many other somatostatin fibers of the frog median eminence originate from somatostatin neurons located outside the tuber cinereum. Most of these neurons probably lie in the preoptic hypothalamic region.
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  • 79
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Corticotrophs ; Immunocytochemistry ; Growth hormone ; Prolactin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two antisera, Y-10 and Y-18 were raised in rabbits against synthetic human β-endorphin conjugated to bovine serum albumin and keyhole limpet haemocyanin respectively. Antiserum Y-10 has been shown by radioimmunoassay to be highly specific for human β-endorphin with minimal or no cross-reactivity against other pituitary peptides whilst antiserum Y-18 crossreacted on an equimolar basis against β-endorphin and β-lipotropin. When used in the immunohistochemical procedure, both antisera specifically stained the corticotrophs in human anterior pituitary tissue. A similar effect was observed when antiserum Y-18 was applied to rat anterior pituitary tissue in the immunohistochemical procedure. Y-10 antiserum, on the other hand, stained not only rat corticotrophs but also somatotrophs. The somatotrophin staining could not be attributed to the enkephalins reported to be present in these cells. The non-specific β-endorphin antiserum Y-18 was used to stain anterior pituitaries from dehydrated and adrenalectomized rats as well as rats of the Brattleboro strain. In tissues from the three experimental animals, cells that stained positively for β-endorphin did not give a positive immunoreaction for ACTH and vice versa in some other sections. It is concluded that under the physiological conditions, formalin fixation of the tissue causes the proopiocortin molecule to be “trapped” in a conformation such that either ACTH or β-endorphin-like determinants are available for reacting with the appropriate antiserum.
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  • 80
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Vasoactive intestinal polypeptide ; Radioimmunoassay ; Gut hormones ; Anura
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical and radioimmunological techniques with region specific antisera have been used to identify a vasoactive intestinal polypeptide-like material in the anuran intestine. Seven species of Anura were investigated: Bombina bombina, Alytes obstetricans, Rana temporaria, Rana esculenta, Hyla arborea, Hyla crepitans and Bufo bufo. In five of the species (A. obstetricans, R. temporaria, H. arborea, H. crepitans and B. bufo) vasoactive intestinal polypeptide-like immunoreactive mucosal endocrine cells and nerve fibres in all layers of the gut wall, were detected by both immunofluorescence and peroxidase-antiperoxidase methods. In the other two species, R. esculenta and B. bombina, no mucosal endocrine cells were detected although the vasoactive intestinal polypeptide-immunoreactive nerve fibres were plentiful. Radioimmunoassay showed the presence of significant amounts of vasoactive intestinal polypeptide-immunoreactivity in intestinal extracts from all species. The highest quantities were present in those anurans with both immunostained cells and nerves. Gel permeation chromatography showed that most of the vasoactive intestinal polypeptide-like peptide eluted in a position identical to that of natural mammalian (porcine) vasoactive intestinal polypeptide. The results indicate that a vasoactive intestinal polypeptide-like peptide is well represented in the Anura and that it is immunologically very similar to the mammalian peptide.
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  • 81
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Substance P ; Chicken gut ; Radioimmunoassay ; Immunocytochemistry ; Smooth muscle motility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution and cellular localization of substance P in the chicken gut was studied by immunocytochemistry and immunochemistry. Substance P-containing nerve fibers are numerous in the gut wall. They occur in the smooth muscle layer as well as in the mucosa, where they are associated with blood vessels or surround the intestinal crypts. The fibers are particularly numerous in the myenteric and submucosal plexuses, where substance P-containing nerve-cell perikarya are also encountered. Substance P was found also in scattered endocrine cells of the small intestine, caeca and colon. Previously, bombesin-containing cells, which are numerous in the proventriculus, have been mistakenly identified as substance P cells due to crossreactivity of certain antisera against substance P. Immunochemistry revealed the highest concentration of substance P in the duodenum. The gel chromatographic behavior of chicken substance P differs slightly from that of synthetic bovine substance P, suggesting that chicken substance P differs structurally from mammalian substance P. Substance P-containing nerve fibers in the chicken gut develop slowly after hatching, apparently beginning in the duodenum; at approximately 20 weeks after hatching the distribution pattern is fully developed. A functional investigation was performed on the isolated chicken caecum to clarify the role of substance P in the contractile behavior of smooth muscle. Substance P contracted the caecum over a wide dose range; the contractile response was greater in 20 week-old chickens than in 4 and 10 week-old animals. Electrical field stimulation caused a relaxation of the caecum and a contraction upon cessation of stimulation. Neither of these responses, both of which are neurally mediated, were inhibited by adrenergic and cholinergic blockade. It is conceivable that the contractile response following electrical stimulation is caused by substance P released from nerve fibers in the smooth muscle.
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  • 82
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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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  • 83
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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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  • 84
    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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  • 85
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    Cell & tissue research 233 (1983), S. 233-236 
    ISSN: 1432-0878
    Keywords: Y-organ ; Steroid secretion ; Ecdysone ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    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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  • 86
    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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  • 87
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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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  • 88
    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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  • 89
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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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  • 90
    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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  • 91
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Supraoptic nucleus (SON) ; Oxytocin neurones ; Neuronal appositions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    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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  • 92
    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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  • 93
    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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  • 94
    ISSN: 1432-0878
    Keywords: Mesotocin neurons ; Vasotocin neurons ; Avian hypothalamus ; Preoptic area ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neurosecretory systems producing mesotocin (MT) and vasotocin (VT) (the avian homologues of oxytocin and vasopressin, respectively) were characterized in the brains of the domestic mallard and Japanese quail by means of indirect immunofluorescence techniques using specific antisera. In the anterior preoptic region, including the organum vasculosum of the lamina terminalis, and at different levels of the supraoptic and paraventricular nuclei, separate mesotocin- and vasotocin-producing neurons were identified. Mesotocinergic and vasotocinergic neurons were also located in the tuberomammillary area, among the ectomammillary tract fibers. The supraoptico-neurohypophysial tract, formed by vasotocin- and mesotocincontaining axons, enters the internal zone of the median eminence and ends in the posterior lobe of the pituitary. The external zone of the rostral median eminence appears to contain vasotocin and mesotocin fibers, which terminate in close contact with the capillaries of the hypophysial portal system.
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  • 95
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; FMRF-amide ; Central nervous system ; Neurohormone ; Neurotransmitter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With an antiserum to the molluscan cardio-excitatory tetrapeptide FMRF-amide neurons and/or nerve fibers were immunocytochemically identified in the central nervous systems of a snail (Lymnaea stagnalis), some insects (Leptinotarsa decemlineata, Periplaneta americana, Locusta migratoria, Pieris brassicae), a fish (Poecilia latipinna) and a mammal (mouse). The fact that immunoreactive material was observed in neurohaemal organs (corpora cardiaca of the insects) as well as in axon terminals ending on other neurons, seems to indicate that this peptide can function as a neurohormone and/or as a neurotransmitter. The results sustain the hypothesis that biologically active peptides have a wide distribution in the animal kingdom.
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  • 96
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 213 (1980), S. 53-68 
    ISSN: 1432-0878
    Keywords: Peptidergic neuroendocrine system ; Domestic fowl ; Metenkephalin system ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the central nervous system of the male domestic fowl, met-enkephalin (ENK) immunoreactive perikarya and fiber tracts as well as extensive but sharply delimited fiber networks were visualized by means of the PAP technique. The most striking results were: (1) The demonstration of an association of ENK-containing structures with branchial nerves; (2) the spatial relationship of ENK-containing perikarya and fibers to somatostatin (SOM) and arginine-vasotocin (AVT)-immunoreactive systems; (3) the presence of dense and extensive ENK fiber networks within (a) the caudo-basal wall of the third ventricle and (b) the septal-preoptic area; in both regions mainly ENK fibers, but also SOM and AVT fibers, may cross to the contralateral side.
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  • 97
    ISSN: 1432-0878
    Keywords: 17–39 ACTH ; LHRH terminals ; Median eminence ; Guinea pig ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary At the ultrastructural level a peptide immunocytochemically related to ACTH was localized in LHRH-containing terminals and nerve fibers of the palisade zone in the median eminence of the guinea pig. Precise comparisons of adjacent ultrathin sections stained for 17–39 ACTH and for LHRH made it possible to state clearly that in the two cases the staining was confined to granules containing both peptides. The significance of such an association remains open to discussion.
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  • 98
    ISSN: 1432-0878
    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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  • 99
    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.
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  • 100
    ISSN: 1432-0878
    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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