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  • Immunohistochemistry  (282)
  • Springer  (282)
  • Annual Reviews
  • Blackwell Publishing Ltd
  • 2020-2024
  • 2005-2009
  • 1990-1994  (185)
  • 1980-1984  (65)
  • 1975-1979  (32)
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  • 1
    ISSN: 1432-0827
    Keywords: Anticollagen antibodies ; Collagen types ; Immunohistochemistry ; Ossified posterior longitudinal ligament
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Immunohistochemical localization of types I, II, and III collagen in the ossified posterior longitudinal ligament of the cervical spine was studied using type-specific anticollagen antibodies. In contrast to the normal ligament which contains both types I and III collagens, the ossified matrix, composed of lamellar bone, contains only type I collagen, except for Haversian canals where type III is located in the inner wall. In the transitional region of preossifying ligaments, types III and I are both present. Type II collagen is present in the hyperplastic matrix of the ligament, and cartilage-like cells surrounded by type II collagen are aligned along nonossified ligaments adjacent to the preossifying region. A possible mechanism of matrix transition during the ossification process is given attention.
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  • 2
    ISSN: 1432-1432
    Keywords: Aphids ; Endosymbiosis ; Symbionin ; Chaperonin 60 ; Chaperonin 10 ; Immunoblotting ; Immunohistochemistry ; Primary symbiont ; Secondary symbiont ; Endosymbiotic evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary All aphids harbor symbiotrophic prokaryotes (“primary symbionts”) in a specialized-abdominal cell, the bacteriocyte. Chaperonin 60 (Cpn60, symbionin) and chaperonin 10 (Cpn10), which are high and low molecular weight heatshock proteins, were sought in tissues of more than 60 aphid species. The endosymbionts were compared immunologically and histologically. It was demonstrated that (1) there are two types of aphids in terms of the endosymbiotic system: some with only primary symbionts and others with, in addition, secondary symbionts; (2) the primary symbionts of various aphids are quite similar in morphology whereas the secondary symbionts vary; and (3) irrespective of the aphid species, Cpn60 is abundant in both the primary and secondary symbionts, while Cpn10 is abundant in the secondary symbionts but present in small amounts in the primary ones. Based on these results, we suggest that the primary symbionts have been derived from a prokaryote that was acquired by the common ancestor of aphids whereas the secondary symbionts have been acquired by various aphids independently after divergence of the aphid species. In addition, we point out the possibility that the prokaryotes under intracellular conditions have been subject to some common evolutionary pressures, and as a result, have come to resemble cell organelles.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 190 (1981), S. 127-131 
    ISSN: 1432-041X
    Keywords: Uteroglobin ; Radioimmunoassay ; Progesterone ; Osmotic minipumps ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This study was undertaken to determine whether the influx of progesterone into the uterine lumen of the rabbit, in the preimplantation period, is dependent onuteroglobin (UGL). Rabbits were ovariectomized and, three months later, treated with two defferent doses of progesterone. Purified UGL was injected into one uterine horn and, as a control,immunoglobulin G (IgG) was injected into the other. After four days, the animals were sacrificed their uteri flushed, and the progesterone content of the washes was determined by radioimmunoassay. Animals with the lower serum progesterone level (2.8 ng/ml) had a significantly different uterine horn progesterone content. The hormone accumulation in the horn containing UGL was 2.3 to 7.5 times higher than in the horn containing IgG. Animals with a higher serum progesterone level (7.2 ng/ml) showed no differences. The hormone content was equally high in both horns, presumably due to the synthesis of endogenous UGL being reactivated by the hormone treatment. The validity of these experiments as models for the events during early pregnancy and the physiological role of progesterone available inside the uterus are discussed.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 24 (1977), S. 73-79 
    ISSN: 1432-0827
    Keywords: Bone cells ; Cyclic GMP ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Guanosine 3′,5′-monophosphate (cyclic GMP, cGMP) was localized in bone cells by the use of an immunoglobulin-enzyme bridge method. We observed that in cat alveolar bone most osteoblasts did not stain for cGMP, while adjacent periodontal cells displayed cytoplasmic as well as nuclear staining. Numerous osteocytes contained diffuse reaction products over most or all of the cellular area. The method used in this study may be helpful in identifying specific hard tissue cell types whose function(s) involve cGMP.
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  • 5
    ISSN: 1432-0827
    Keywords: Decorin ; Proteoglycan-100 ; Heterotopic ossification ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Heterotopic ossification is a metabolically active process which shares several properties of orthotopic bone formation and, therefore, represents an excellent model for studying bone matrix components. Immunohistochemical methods were used to investigate the distribution pattern of the small proteoglycans decorin and proteoglycan-100 during different stages of heterotopic ossification of pressure sores of paraplegic patients. Decorin and proteoglycan-100 exhibited a substantially divergent distribution pattern. Decorin was detectable in the perivascular matrix of granulation tissue as well as in the stroma of heterotopic ossification. The ossification zone was stained most strongly. In contrast, proteoglycan-100 was predominantly detectable in fibroblasts and preosteoblasts in early areas of osteogenesis. In more mature forms of heterotopic ossification immunostaining was markedly reduced in osteoblasts and osteocytes and even absent in so-called bone-lining cells. However, at least some osteoclasts were strongly positive. These results suggest indicate that decorin and proteoglycan-100 are important components during the formal pathogenesis of heterotopic ossification. The expression of the small proteoglycans, especially of proteoglycan-100, correlates with different phases during heterotopic ossification, showing a maximum for proteoglycan-100 in matrix-forming cells in early phases of bone formation, but in osteoclasts in mature bone.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 55 (1994), S. 38-45 
    ISSN: 1432-0827
    Keywords: Vacuolar-type H+-ATPase ; Carbonic anhydrase II ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The localization of vacuolar-type H+-ATPase and carbonic anhydrase II (CA II) in rat incisor enamel organs at maturation was examined by light and electron microscopy. The immunoreactivity for both vacuolar-type H+-ATPase and CA II was intense on the ruffled border of ruffle-ended ameloblasts (RA), but moderate at the distal end of smooth-ended ameloblasts (SA). Immuno-gold particles indicated that CA II was not confined to the ruffled border of RA alone, but also distributed in the cytoplasm of RA and SA. These findings suggest that RA may secrete protons produced by CA II via the ruffled border into enamel by active transport of vacuolar-type H+-ATPase. Secreted protons may activate hydrolytic enzymes to degrade the organic components of enamel matrix. Vacuolar-type H+-ATPase on vesicles of SA suggests that a specific configuration of ruffled borders in RA may be formed by the fusion of vesicle membranes in the distal end of cytoplasm of SA.
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  • 7
    ISSN: 1432-1254
    Keywords: Temporal variation ; Cytochrome P-450 isozyme ; Testis ; Immunohistochemistry ; Cadinenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geography , Physics
    Notes: Abstract Temporal variations in cytochrome P-450 isozymes of rat testis, PB-P-450 (forms of cytochrome P-450 strongly induced by phenobarbital) and MC-P-448 (forms of cytochrome P-450 strongly induced by 3-methylcholanthrene), were investigated immunohistochemically by the avidin-biotin-complex method using specific antibodies against PB-P-450 and MC-P-448 isozymes. Immunoreactivity to both PB-P-450 and MC-P-448 isozymes was observed in Leydig cells. The number of PB-P-450 positive Leydig cells was found to undergo significant time-of-day variation with a peak time of 0000 hours (light phase from 0800 to 2000 hours). Injection of cadinenes (300 mg/kg per day intraperitoneally at 48 and 96 h before sacrifice) induced PB-P-450 isozyme but did not induce MC-P-448 isozyme. The induction of PB-P-450 isozyme by cadinenes was time dependent, and the early dark phase (2000 and 0000 hours) was most sensitive. These results suggest that temporal variation of cytochrome P-450 isozymes is one of the important physiological variations in detoxification and activation of various xenobiotics and chemicals in the testis.
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  • 8
    ISSN: 1432-0878
    Keywords: Key words: NADPH diaphorase ; Nitric oxide synthase ; Nervous system ; central ; Nervous system ; peripheral ; Immunohistochemistry ; Helix aspersa (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Enzyme histochemistry and immunocytochemistry were used to determine the distribution of neurons in the snail Helix aspersa which exhibited nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and/or immunoreactivity to nitric oxide synthase (NOS). NADPH diaphorase-positive cells and fibres were distributed extensively throughout the central and peripheral nervous system. NADPH diaphorase-positive fibres were present in all neuropil regions of the central and peripheral ganglia, in the major interganglionic connectives and in peripheral nerve roots. NADPH diaphorase-positive cell bodies were found consistently in the eyes, the lips, the tentacular ganglia and the procerebral lobes of the cerebral ganglia; staining of cell bodies elsewhere in the nervous system was capricious. The distribution of NOS-like immunoreactivity differed markedly from that of NADPH diaphorase activity. Small clusters of cells which exhibited NOS-like immunoreactivity were present in the cerebral and pedal ganglia; fibres which exhibited NOS-like immunoreactivity were present in restricted regions of the neuropil of the central ganglia. The disjunct distributions of NADPH diaphorase activity and NOS-like immunoreactivity in the nervous system of Helix suggest that the properties of neuronal NOS in molluscs may differ sigificantly from those described previously for vertebrate animals.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 277 (1994), S. 457-464 
    ISSN: 1432-0878
    Keywords: Mammary-derived growth inhibitor ; Fatty acid-binding proteins ; Differentiation ; Vascularization ; Immunohistochemistry ; Immunocytochemistry ; Mammary gland ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mammary-derived growth inhibitor (MDGI) has previously been localized in the mammary parencyma, dependent on the stage of differentiation of the mammary gland. Here, we have elucidated the distribution of MDGI in the mammary stroma by a combined immunohisto-and cytochemical analysis with antibodies raised against MDGI. Distinct staining of capillary endothelial cells has been revealed. Although its subcellular distribution resembles former observations in secretory epithelial cells, the expression of MDGI in capillary endothelial cells clearly precedes that in secretory epithelial cells. On the other hand, no endothelial MDGI staining has been detected in bovine heart, which contains a fatty acid-binding protein almost identical to MDGI. The localization of MDGI in the mammary capillary endothelium is discussed in terms of its possible involvement in the intracellular transport of hydrophobic ligands or in the regulation of endothelial cell proliferation.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 156 (1975), S. 167-171 
    ISSN: 1432-0878
    Keywords: Pancreatic islet cells ; Fourth cell type ; Human pancreatic polypeptide ; Localization ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary HPP is the human counterpart of APP—a polypeptide that has been isolated from avian pancreas and shown to exert pronounced biological effects. The availability of anti-HPP serum enabled us to use immunohistochemical procedures for the localization of HPP in human pancreas. A small population of pancreatic islet cells showed strong immunofluorescence after staining with anti-HPP serum. The HPP cells were mainly localized at the periphery of the islets. Sometimes they were also seen scattered in the exocrine parenchyma as well as within the epithelium of small to medium-sized ducts. While rare in the pancreas of adult man, HPP cells were abundant in certain parts of the pancreas of foetuses (18–20 weeks gestational age). Their staining properties showed them to be distinct from the A, B and D cells of the pancreatic islets.
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  • 11
    ISSN: 1432-0878
    Keywords: Key words: Chromogranins ; Serotonin ; Amylin ; Regulatory peptides ; Gut ; Monoclonal antibodies ; Immunohistochemistry ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Novel monoclonal antibodies to human chromogranin A (CgA) and chromogranin B (CgB) were used to investigate the presence of immunoreactive (-IR) elements in the alimentary tract of the green frog Rana esculenta. Numerous CgA-IR and a few CgB-IR endocrine cells were found within the gut mucosa, from the oesophagus to the cloaca, with some local differences in density. Co-localization studies demonstrated that they were co-stored in almost all the serotonin-IR, the amylin-IR or islet amyloid polypeptide-IR cells and in the peptide tyrosine tyrosine-IR cells located proximal to the pylorus, but not in those located in more caudal tracts. No other co-localization was demonstrated; substances investigated included somatostatin, substance P, gastrin/cholecystokinin, glucagon, glycentin, bombesin, secretin and neurotensin. CgA-IR and CgB-IR cells nearly always displayed argyrophilia with the Grimelius silver method
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  • 12
    ISSN: 1432-0878
    Keywords: Somatostatin ; Immunohistochemistry ; Neuroendocrine function
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Utilizing the unlabeled antibody peroxidase-antiperoxidase method, we have studied the distribution of somatostatin (growth hormone release-inhibiting hormone) in the guinea pig brain. An immunohistochemical reaction was found in both internal and external zone of the caudal and median portions of the median eminence. Reaction products were also present in the arcuate nucleus in proximity of the third ventricule. The immunostained elements resembled nerve processes. No cell bodies were stained. A positive reaction was also present in the whole organum vasculosum of the lamina terminalis. In the subfornical and subcommissural organs, somatostatin was observed in the ependymal and subependymal layers and the perivascular areas. In the pineal gland, a positive reaction was also found mainly in the perivascular areas. All the other brain areas were completely negative. These results establish that, in the hypothalamus, somatostatin is mainly concentrated in the median eminence and suggest that periventricular organs can play a role in the regulation of growth hormone secretion.
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  • 13
    ISSN: 1432-0878
    Keywords: Chromogranins ; Serotonin ; Amylin ; Regulatory peptides ; Gut ; Monoclonal antibodies ; Immunohistochemistry ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Novel monoclonal antibodies to human chromogranin A (CgA) and chromogranin B (CgB) were used to investigate the presence of immunoreactive (-IR) elements in the alimentary tract of the green frog Rana esculenta. Numerous CgA-IR and a few CgB-IR endocrine cells were found within the gut mucosa, from the oesophagus to the cloaca, with some local differences in density. Co-localization studies demonstrated that they were costored in almost all the serotonin-IR, the amylin-IR or islet amyloid polypeptide-IR cells and in the peptide tyrosine tyrosine-IR cells located proximal to the pylorus, but not in those located in more caudal tracts. No other co-localization was demonstrated; substances investigated included somatostatin, substance P, gastrin/cholecystokin, glucagon, glycentin, bombesin, secretin and neurotensin. CgA-IR and CgB-IR cells nearly always displayed argyrophilia with the Grimelius silver method
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  • 14
    ISSN: 1432-0878
    Keywords: Key words: Corpuscles of Stannius ; Stanniocalcin ; Immunocytochemistry ; Immunohistochemistry ; Western blot ; Lepisosteus osseus (Holostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Stanniocalcin-immunoreactive cells were localized in the corpuscles of Stannius of a holostean fish, the garpike (Lepisosteus osseus), using antisera against salmon and trout stanniocalcins and the peroxidase-antiperoxidase and protein A-gold immunohistochemical methods. The stanniocalcin-immunoreactive cells were periodic acid-Schiff-positive, and antibody staining was abolished if the antiserum was preabsorbed with corpuscle homogenate. Immunocytochemistry revealed two reactive cell types in the glandular parenchyma, and immunoreactivity was confined to the secretory granules. Staining of the granules was also abolished when the antisera were blocked with crude corpuscle homogenate. When corpuscle extracts from garpike were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot analysis, a single dense band was evident with a molecular weight of ∼68 kDa under non-reducing conditions, whereas three bands were observed (∼29, ∼31, and ∼34 kDa) under reducing conditions. Staining of all bands disappeared following preabsorption of the antiserum with salmon stanniocalcin, trout stanniocalcin, or garpike corpuscle extract. The results are compared with stanniocalcins from another extant holostean, the bowfin (Amia calva), and from more modern bony fishes, the teleosts.
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  • 15
    ISSN: 1432-0878
    Keywords: Corpuscles of Stannius ; Stanniocalcin ; Immunocytochemistry ; Immunohistochemistry ; Western blot ; Lepisosteus osseus (Holostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Stanniocalcin-immunoreactive cells were localized in the corpuscles of Stannius of a holostean fish, the garpike (Lepisosteus osseus), using antisera against salmon and trout stanniocalcins and the peroxidase-antiperoxidase and protein A-gold immunohistochemical methods. The stanniocalcin-immunoreactive cells were periodic acid-Schiff-positive, and antibody staining was abolished if the antiserum was preabsorbed with corpuscle homogenate. Immunocytochemistry revealed two reactive cell types in the glandular parenchyma, and immunoreactivity was confined to the secretory granules. Staining of the granules was also abolished when the antisera were blocked with crude corpuscle homogenate. When corpuscle extracts from garpike were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot analysis, a single dense band was evident with a molecular weight of ∼68 kDa under non-reducing conditions, whereas three bands were observed (∼29, ∼31, and ∼34 kDa) under reducing conditions. Staining of all bands disappeared following preabsorption of the antiserum with salmon stanniocalcin, trout stanniocalcin, or garpike corpuscle extract. The results are compared with stanniocalcins from another extant holostean, the bowfin (Amia calva), and from more modern bony fishes, the teleosts.
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  • 16
    Electronic Resource
    Electronic Resource
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    Cell & tissue research 277 (1994), S. 325-331 
    ISSN: 1432-0878
    Keywords: c-Myc ; c-Fos ; Vasoactive intestinal polypeptide (VIP) ; Immunohistochemistry ; Intestine, small ; Enteric nervous system ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical double labelling of the enteric nervous system of the guinea pig ileum was performed with a monoclonal antibody (anti-MYC 033) directed against a peptide sequence of the human c-Myc protein together with antibodies directed against either the neuron-specific antigens neuron-specific enolase or PGP 9.5 or the glia-specific marker S-100 to demonstrate that anti-MYC 033 labelled the nuclei of all enteric neurons but not glia. This strategy was also employed to demonstrate that another anti-c-Myc monoclonal anti-body, anti-MYC 070, labelled the nuclei of all neurons and glia, as well as perhaps all other cells in these preparations. A polyclonal antiserum raised against a peptide sequence of the human c-Fos protein (anti-FOS 4) was shown to label the identical nuclei as anti-MYC 033. The ganglionic density of nuclei labelled by anti-FOS 4 was found to be similar to previous measures of the ganglionic density of neurons. Double labelling with anti-MYC 033 and an antiserum directed against vasoactive intestinal polypeptide was performed to reexamine the ganglionic density of neurons that express this neuropeptide. Our results suggest that the ganglionic density of these neurons might be less than previously determined.
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  • 17
    Electronic Resource
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    Cell & tissue research 170 (1976), S. 415-424 
    ISSN: 1432-0878
    Keywords: Uterine specific proteins ; Oviduct ; Uterus ; Blastocyst of the Rabbit ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization and release of uteroglobin (UGL) were investigated immunohistologically in the oviducts and uteri of female rabbits from oestrus through the 7th day post coitum and the blastocyst on the 7th day post coitum. UGL was detected within Fallopian tube cells even during oestrus. Granules of UGL appeared toward the bases of these cells. Subsequently, the cells became almost entirely filled with UGL. Drop-like protrusions of the apical cytoplasm suggest a mechanism of apocrine extrusion. All stages of filling and extrusion were visible during the entire preimplantation period. During oestrus, synthesis of UGL within uterine cells becomes sufficiently advanced so that extrusion has either already begun or is about to begin. UGL positive material first appears in the supranuclear regions. Later the entire cytoplasm shows a positive reaction. An uneven distribution of UGL cells is observed in the endometrium. Since only the glands adjacent to the myometrium and cells of the cavum epithelium contain UGL, a striking mosaic of UGL positive and negative cells results. The present report is the first detecting UGL in single cells of the blastocyst. Both entodermal and ectodermal cells proved to be UGL positive. The synthesis and section of UGL in the oviduct and uterus and the possible origins of UGL in the blastocyst are discussed.
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  • 18
    Electronic Resource
    Electronic Resource
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    Cell & tissue research 170 (1976), S. 425-434 
    ISSN: 1432-0878
    Keywords: Uterine specific proteins ; Uterus of the rabbit ; Immunohistochemistry ; Oestrogen-Progesterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rabbit uterine uteroglobin (UGL) was studied by electrophoretic and immunological methods following normal copulation, after ovariectomy and progesterone treatment, 17β-oestradiol and combined progesterone treatment, 17β-oestradiol treatment alone and after HCG-induced pseudopregnancy. Electrophoretic studies show the amount of ULG in uterine secretions, the immunological investigations indicate the intracellular localization of ULG and the distribution of ULG-positive cells in the endometrium. No obvious differences were found between the uteri 7 days after injection with chorion-gonadotropin and those 7 days following normal copulation. No differences could be demonstrated between the uteri of animals 35 days following ovariectomy and subsequent progesterone treatment on Days 31–33 and those of normal 7 d. post coitum (p.c.) animals. Uteri from animals treated with progesterone on Days 2–5 p.c. contained more ULG-positive cells than controls. 17β-oestradiol treatment with and without subsequent progesterone treatment resulted, in both gravid and ovariectomized animals, in the formation of a tall columnar endometrial epithelium. Treatment with 17β-oestradiol on Days 1 and 2 p.c. led to a decrease in the number of UGL-positive cells at 7 days p.c. Even after ovariectomy with 17β-oestradiol substitution, UGL-positive cells were still present in the endometrium. However a secretion of any magnitude could not be detected. The importance of differentiation between synthesis and secretion (= release) as distinct phases of the glandular response is especially emphasised by the latter findings.
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  • 19
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Japanese quail ; Immunohistochemistry ; LH producing cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cells that produce luteinizing hormone (LH) in the adenohypophysis of the Japanese quail were identified immunohistochemically using anti-chicken LH serum and horseradish peroxidase-labeled goat anti-rabbit gamma globulin serum. The LH cells are localized in the caudal lobe of the pars distalis. They are elongate in shape and are polarized toward the sinusoids, especially in their active states. Alterations in size of LH cells are directly related to changes in circulating LH levels as induced by castration or photostimulation. The LH cells identified immunohistochemically were only stained by alcian blue with periodic acid-Schiff (PAS), alcian blue and orange G. PAS-positive gonadotropic cells in the cephalic lobe were stained immunohistochemically only slightly if at all using anti-chicken LH serum and consequently may be FSH producing cells. In the cephalic lobe another type of basophilic cell was stained with alcian blue. These cells were also stained immunohistochemically with anti-chicken LH serum. These cells may possibly be identified as TSH cells due to the characteristics of the antichicken LH serum used in this study which cross react with LH and TSH but only slightly with FSH, and also on the basis of previous light and electron microscopic studies.
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  • 20
    Electronic Resource
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    Cell & tissue research 166 (1976), S. 407-412 
    ISSN: 1432-0878
    Keywords: Pituitary primordium ; Cytodifferentiation ; Rat ; Organ culture ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rathke's pouches isolated from rat fetuses on day 12 were maintained in organ culture for 9 days and investigated immunohistochemically to test whether or not the hypothalamus is involved in the cytodifferentiation of the adenohypophysis. The unlabeled antibody enzyme method demonstrated that the cultured tissue contains different types of glandular cells, i.e., adrenocorticotropin (ACTH)-, growth hormone (GH)-, luteinizing hormone (LH)-, thyrotropin (TSH)-, and prolactin-producing cells. Indirect evidence was also obtained to indicate the presence of melanocyte stimulating hormone (MSH)-cells. These findings suggest that adenohypophysial primordial cells of rats start to synthesize their respective hormones without stimuli from neurosecretory substances of the brain which are known to be essential for the maintenance of the secretory activity of the adult gland.
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  • 21
    Electronic Resource
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    Cell & tissue research 200 (1979), S. 83-90 
    ISSN: 1432-0878
    Keywords: Spinal cord ; Tupaia ; Somatostatin ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Somatostatinergic nerves in the spinal cord of the monkey were investigated utilizing immunohistochemistry with various antibodies against synthetic somatostatin. In contrast to earlier investigations, it is shown that somatostatinergic nerve endings occur in most of the areas of the grey matter of the spinal cord. The somatostatinergic axons are, however, characteristically distributed in three main regions: (1) Densely-packed endings are seen in lamina II of the substantia gelatinosa, forming a crescent-shaped pattern in the columna dorsalis. Somatostatin immunoreactivity is also seen in lamina I and in the Lissauer tract. (2) A fine network of fibers is observed around the central canal; the endings are concentrated on special cell bodies. Some single perikarya are also stained in this region. (3) A loose network of single fibers is found ending on perikarya of the columna lateralis or ventralis. The perikarya of the nerve axons, with the exception of those terminating in the columna dorsalis, have as yet not been identified. In order to better understand the somatostatinergic system of the spinal cord, these newly-detected somatostatinergic nerves must be studied and their exact pathways analyzed.
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  • 22
    Electronic Resource
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    Cell & tissue research 277 (1994), S. 325-331 
    ISSN: 1432-0878
    Keywords: Key words: c-Myc ; c-Fos ; Vasoactive intestinal polypeptide (VIP) ; Immunohistochemistry ; Intestine ; small ; Enteric nervous system ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical double labelling of the enteric nervous system of the guinea pig ileum was performed with a monoclonal antibody (anti-MYC 033) directed against a peptide sequence of the human c-Myc protein together with antibodies directed against either the neuron-specific antigens neuron-specific enolase or PGP 9.5 or the glia-specific marker S-100 to demonstrate that anti-MYC 033 labelled the nuclei of all enteric neurons but not glia. This strategy was also employed to demonstrate that another anti-c-Myc monoclonal antibody, anti-MYC 070, labelled the nuclei of all neurons and glia, as well as perhaps all other cells in these preparations. A polyclonal antiserum raised against a peptide sequence of the human c-Fos protein (anti-FOS 4) was shown to label the identical nuclei as anti-MYC 033. The ganglionic density of nuclei labelled by anti-FOS 4 was found to be similar to previous measures of the ganglionic density of neurons. Double labelling with anti-MYC 033 and an antiserum directed against vasoactive intestinal polypeptide was performed to reexamine the ganglionic density of neurons that express this neuropeptide. Our results suggest that the ganglionic density of these neurons might be less than previously determined.
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  • 23
    ISSN: 1432-0878
    Keywords: Central nervous system ; Neuropeptides ; Immunohistochemistry ; Amphioxus, Branchiostoma belcheri (Cephalochorda)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunohistochemical localization of nine different neuropeptides was studied in the central nervous system of the amphioxus, Branchiostoma belcheri. In the brain, perikarya immunoreactive for urotensin I and FMRFamide were localized in the vicinity of the central canal. One of the processes of each of these perikarya was found to cross the dorso ventral slit-like lumen of the central canal. Oxytocin-immunoreactive short fibers, but not perikarya, were detected in the ventral part of the brain. Perikarya immunoreactive for arginine vasopressin/vasotocin, oxytocin and FMRFamide were widely distributed in the spinal cord. Arginine vasopressin/vasotocin-immunoreactive fibers often made contacts with Rohde cell axons. Angiotensin II-immunoreactive perikarya were observed in the posterior half of the spinal cord, and urotensin I-immunoreactive perikarya were found in the caudal region of the spinal cord. Cholecystokinin/gastrin-immunoreactive fibers, but not perikarya, were detected in the spinal cord; some extended as far as the ependymal layer of the cerebral ventricle. No colocalization of the peptides examined was observed. No immunoreactivity for atrial and brain natriuretic peptides nor for urotensin II was detected. The present study indicates that there are at least six separate neuronal systems that contain different peptides, respectively, in the central nervous system of the amphioxus. Their functions remain to be determined.
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  • 24
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    Cell & tissue research 176 (1977), S. 529-551 
    ISSN: 1432-0878
    Keywords: Pancreatic islets ; Correlative light and electron microscopy ; Immunohistochemistry ; Insulin ; Glucagon ; Xiphophorus helleri
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé Cinq catégories cellulaires sont décrites dans l'îlot endocrine du poisson téléostéen Xiphophorus helleri. Chacune est définie par un ensemble de caractères histochimiques et ultrastructuraux, ce qui peut être fait avec certitude grâce à l'étude comparée de coupes ultra-fines et sémi-fines successives. Les techniques d'immunofluorescence ont été appliquées à du matériel préparé pour la microscopie électronique, à l'aide de sérums anti-glucagon et anti-insuline de porc. Un des résultats les plus intéressants est la démonstration de l'existence de 2 catégories de cellules A2: les cellules Adams positives, qui ont des grains de sécrétion amorphes et de section circulaire (≪ cellules à grains ronds ≫) et les cellules Adams négatives, dont de nombreux grains sont des cristaux (≪cellules à grains cristallins≫). Les cellules B et les cellules A1 constituent deux autres types cellulaires. Les ≪cellules claires≫, qui ne réagissent à aucune des techniques employées, représentent un type cellulaire distinct des précédents. L'influence de la fixation se révèle très importante, aussi bien sur les affinités tinctoriales des cellules que sur leurs caractères ultrastructuraux. The authors are grateful to Mrs. L.G. Heding, Novo Industri A/S, Bagsvaerd, for antiporcine insulin and glucagon sera, to Dr. S. Syed Ali, Elektronenmikroskopie, Zentrum für Anatomie und Cytobiologie, Universität Giessen, for preparing FITC-labeled γ-globulin, and to Mr. D. Streicher, Laboratoire de Zoologie et Embryologie expérimentale, Université Louis Pasteur, Strasbourg, for technical assistance
    Notes: Summary By means of correlative light and electron microscopy, five pancreatic islet cell categories are described in the teleost fish, Xiphophorus helleri, each of which has specific light microscopic appearance and fine structure. Different histochemical techniques have been used, including immunofluorescence with antiporcine insulin and glucagon sera. In addition to B- and A1-cells, two categories of A2-cells have been observed, both reacting with antiporcine glucagon serum: A2-cells with round granules gave a positive reaction for tryptophan; A2-cells with crystalline granules gave a negative reaction with the same staining technique on the same section. The “clear cells”, the last category, were not specifically stained by any of the staining methods carried out in this investigation. The influence of fixation on staining affinities and on ultrastructure was shown to be considerable.
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  • 25
    ISSN: 1432-0878
    Keywords: NADPH diaphorase ; Immunohistochemistry ; Gastrointestinal tract ; Nitric oxide ; Histochemistry ; Guinea-pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and abundance of nitric oxide synthase (NOS)-containing neurons and their terminals in the gastrointestinal tract of the guinea-pig were examined in detail using NADPH diaphorase histochemistry and NOS immunohistochemistry. NOS-containing cell bodies were found in the myenteric plexus throughout the gastrointestinal tract and in the submucous plexus of the stomach, colon and rectum. NOS-containing neurons comprised between 12% (in the duodenum) and 54% (in the esophagus) of total myenteric neurons. In the ileum, NOS neurons represented 19% of total myenteric neurons. Most of the NOS neurons throughout the gastrointestinal tract possessed lamellar dendrites and a single axon. NOS-containing terminals were abundant in the circular muscle, including that of the sphincters, but were rare in the longitudinal muscle, except for the taeniae of the caecum. The muscularis mucosae of the esophagus, stomach, colon and rectum received a medium to dense innervation by NOS terminals. Within myenteric ganglia, NOS-containing terminals were extremely sparse in the esophagus, stomach and duodenum, common in the ileum and distal colon and extremely dense in the proximal colon and rectum. The submucous plexus in the ileum and large intestine contained a sparse plexus of NOS-containing terminals. NOS terminals were not observed in the mucosa of any region. We conclude that throughout the gastrointestinal tract of the guinea-pig, NOS neurons are inhibitory motor neurons to the circular muscle; in the ileum and large intestine, NOS neurons may also function as interneurons.
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  • 26
    ISSN: 1432-0878
    Keywords: Substance P ; Calcitonin gene ; related peptide ; Cervical spinal nerve ; Immunohistochemistry ; Primary afferents ; Mouse (ICR)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We demonstrate the existence of nerve fibers possessing substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactivity in the mouse cervical ventral roots. The distribution of the SP and CGRP fibers was similar, but CGRP fibers were generally more numerous. Both types entered the ventral pia mater or formed hairpin loops, but they did not enter the spinal cord directly through these roots. SP and CGRP fibers in the ventral roots were thin and had many varicosities. We suggest that these SP and CGRP fibers are involved not only in a sensory mechanism, but also in other functions, via the release of SP and CGRP from varicosities in the ventral roots.
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  • 27
    ISSN: 1432-0878
    Keywords: Key words: Pituitary ; Gonadotrophs ; LHβ ; FSHβ ; Immunohistochemistry ; Rana japonica (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical localization of lutropin β (LHβ) and follitropin β (FSHβ) in the pituitary gland of the frog Rana japonica was studied by the peroxidase-anti-peroxidase method and the two-face, double-labeling method with different-sized gold particles at the light- and electron-microscopic levels, respectively, using monoclonal antibodies against bullfrog LHβ and FSHβ. Light-microscopic immunohistochemistry indicated that approximately 66.0% of all the gonadotrophs in the pituitary contained both LHβ and FSHβ, whereas 33.4% of gonadotrophs contained only LHβ, and 0.6% contained only FSHβ. The staining intensity of LHβ and FSHβ varied from cell to cell. The gonadotrophs were classified into four types (Types I-IV) in terms of their ultrastructural and immunolabeling characteristics. Moreover, several secretory granule types were recognized according to differences in their shape and electron density. In all the cell types, both LHβ and FSHβ were often seen in the same secretory granules, but the proportion of granules bearing both hormones ranged from 5.5% in Type I to 32.7% in Type IV. Most secretory granules in Types I and II were immunolabeled with LHβ alone, whereas a small number of granules were immunolabeled with FSHβ alone. More immunolabeled FSHβ granules were present in Types III and IV than in Types I and II.
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  • 28
    ISSN: 1432-0878
    Keywords: NADPH diaphorase ; Nitric oxide synthase ; Nervous system, central ; Nervous system, peripheral ; Immunohistochemistry ; Helix aspersa (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Enzyme histochemistry and immunocytochemistry were used to determine the distribution of neurons in the snail Helix aspersa which exhibited nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and/or immunoreactivity to nitric oxide synthase (NOS). NADPH diaphorase-positive cells and fibres were distributed extensively throughout the central and peripheral nervous system. NADPH diaphorase-positive fibres were present in all neuropil regions of the central and peripheral ganglia, in the major interganglionic connectives and in peripheral nerve roots. NADPH diaphorase-positive cell bodies were found consistently in the eyes, the lips, the tentacular ganglia and the procerebral lobes of the cerebral ganglia; staining of cell bodies elsewhere in the nervous system was capricious. The distribution of NOS-like immunoreactivity differed markedly from that of NADPH diaphorase activity. Small clusters of cells which exhibited NOS-like immunoreactivity were present in the cerebral and pedal ganglia; fibres which exhibited NOS-like immunoreactivity were present in restricted regions of the neuropil of the central ganglia. The disjunct distributions of NADPH diaphorase activity and NOS-like immunoreactivity in the neurvous system of Helix suggest that the properties of neuronal NOS in molluscs may differ sigificantly from those described previously for vertebrate animals.
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  • 29
    ISSN: 1432-0878
    Keywords: Pituitary ; Gonadotrophs ; LHβ ; FSHβ ; Immunohistochemistry ; Rana japonica (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical localization of lutropin β (LHβ) and follitropin β (FSHβ) in the pituitary gland of the frog Rana japonica was studied by the peroxidase-anti-peroxidase method and the two-face, double-labeling method with different-sized gold particles at the light-and electron-microscopic levels, respectively, using monoclonal antibodies against bullfrog LHβ and FSHβ. Light-microscopic immunohistochemistry indicated that approximately 66.0% of all the gonadotrophs in the pituitary contained both LHβ and FSHβ, whereas 33.4% of gonadotrophs contained only LHβ, and 0.6% contained only FSHβ. The staining intensity of LHβ and FSHβ varied from cell to cell. The gonadotrophs were classified into four types (Types I–IV) in terms of their ultrastructural and immunolabeling characteristics. Moreover, several secretory granule types were recognized according to differences in their shape and electron density. In all the cell types, both LHβ and FSHβ were often seen in the same secretory granules, but the proportion of granules bearing both hormones ranged from 5.5% in Type I to 32.7% in Type IV. Most secretory granules in Types I and II were immunolabeled with LHβ alone, whereas a small number of granules were immunolabeled with FSHβ alone. More immunolabeled FSHβ granules were present in Types III and IV than in Types I and II.
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  • 30
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    Cell & tissue research 277 (1994), S. 457-464 
    ISSN: 1432-0878
    Keywords: Key words: Mammary-derived growth inhibitor ; Fatty acid-binding proteins ; Differentiation ; Vascularization ; Immunohistochemistry ; Immunocytochemistry ; Mammary gland ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Mammary-derived growth inhibitor (MDGI) has previously been localized in the mammary parenchyma, dependent on the stage of differentiation of the mammary gland. Here, we have elucidated the distribution of MDGI in the mammary stroma by a combined immunohisto- and cytochemical analysis with antibodies raised against MDGI. Distinct staining of capillary endothelial cells has been revealed. Although its subcellular distribution resembles former observations in secretory epithelial cells, the expression of MDGI in capillary endothelial cells clearly precedes that in secretory epithelial cells. On the other hand, no endothelial MDGI staining has been detected in bovine heart, which contains a fatty acid-binding protein almost identical to MDGI. The localization of MDGI in the mammary capillary endothelium is discussed in terms of its possible involvement in the intracellular transport of hydrophobic ligands or in the regulation of endothelial cell proliferation.
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  • 31
    ISSN: 1432-0878
    Keywords: Vasopressin ; Immunohistochemistry ; Hypothalamus ; Hypophysis ; Murids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Vasopressin was immunohistochemically localized at the electron microscopic (EM) level in the hypothalamic-neurohypophysial system (HNS) of three murids. Antiserum to vasopressin was produced in rabbits injected with lysine vasopressin (LVP) conjugated to egg albumin (EA), anti-EA being precipitated prior to staining. Sternberger's unlabeled antibody peroxidase technique was employed, immunoreactivity being designated by peroxidase anti-peroxidase (PAP) molecules and electron opacity. Immunoreactive neurosecretory granules (NSG) were found in the perikarya of the supraoptic nucleus (SON) in all three murids investigated, although far more profusely in the two wild strains. Immunoreactive axonal NSG were observed in the inner and outer zones of the median eminence (ME), and within most of the axons and terminals in the neurohypophysis. The concentration of primary serum effective for staining the SON (1∶10–1∶50) was far higher than that required for the ME and the neurohypophysis (1:500–1:1,200). AntiLVP also induced electron opacity of granules in cells of the pars intermedia (PI). Discussion centers on the significance of immunoreactive NSG in the neurosecretory (NS) perikarya, on the possibility of an extragranular pool of hormone, and on speculation about the electron opacity of the PI granules.
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  • 32
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    Cell & tissue research 172 (1976), S. 447-454 
    ISSN: 1432-0878
    Keywords: Pancreas ; Horse ; Immunohistochemistry ; Insulin ; Glucagon ; Somatostatin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Equine pancreas was investigated with immunohistochemical methods to study the distribution of endocrine cells immunoreactive to antiinsulin, anti-glucagon, and anti-somatostatin. A-cells demonstrable by antiglucagon are located in the center of Langerhans islets and frequently in the duct epithelium. Few A-cells are seen associated to acini. Anti-insulin reactive B-cells form a large zone around the center of the Langerhans islets in which some B-cells lie between exocrine cells and others, although few, are located in the duct epithelium. D-cells stained with anti-somatostatin serum form a discontinuous outermost zone around the Langerhans islets. In some islets the D-cells are also observed among the B-cells or between the border of A- and B-cells. Single D-cells are seen in the duct epithelium or between acinar cells. In younger horses, endocrine cells are more frequently associated in bulges of the duct system. The histotopographic relation between these endocrine cell types is discussed with respect to its functional significance.
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  • 33
    ISSN: 1432-0878
    Keywords: LH-RH neurons ; Hypothalamus ; Rat ; Immunohistochemistry ; Radioimmunoassay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of the perikarya of LH-RH neurons in the rat hypothalamus and their pathways to the median eminence were studied by immunohistochemistry and radioimmunoassay after placing stereotaxic electrolytic lesions in several parts of the hypothalamus. The principal location of the cell somata was found to be in the ventral part of the medial preoptic area; their pathways were classified into a main baso-lateral pathway and an accessory descending pathway branching off from the former. The main pathway was found to cross in the vicinity of the corresponding neuronal perikarya. The central median eminence and the dorsal and ventral walls of the tubero-infundibular sulcus of the caudal part of the median eminence are innervated mainly by the baso-lateral pathway. On the other hand, the rostral and most caudal portions of the median eminence are innervated principally by the descending pathway and have a subsidiary dual innervation. The projection of LH-RH neurons to the OVLT is believed to originate from perikarya adjacent to this circumventricular organ.
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  • 34
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    Cell & tissue research 235 (1984), S. 703-705 
    ISSN: 1432-0878
    Keywords: Enterochromaffin cells ; Serotonin ; Immunohistochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphology of enterochromaffin (EC) cells in the human fundic mucosa was investigated at the lightmicroscopic level by means of the unlabeled peroxidase anti-peroxidase method, with the use of a highly specific anti-serotonin serum. EC-cells in the human fundic mucosa were sparsely distributed below the neck portion of the gland, but were found to be rather numerous in its lower half. Immunohistochemistry revealed marked pleomorphic and seemingly polynuclear EC-cells or cells with long, sometimes multipolar cytoplasmic processes. In addition, luminal contacts and contiguity between EC-cells, or interglandular connections were also encountered. The present immunohistochemical procedure permits, for the first time, a clear-cut morphological visualization of the entire population of EC-cells, and reveals the distinctive morphological features of these cells in the human fundic mucosa. These morphological findings imply that EC-cells in the fundic mucosa may be crucial in gastric function.
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  • 35
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    Cell & tissue research 237 (1984), S. 239-244 
    ISSN: 1432-0878
    Keywords: Islet A cell ; Catecholamine ; Combined microscopy ; Immunohistochemistry ; Fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to identify pancreatic islet cells emitting formaldehyde-induced fluorescence (FIF), the pancreatic islets of the domestic fowl were studied by combined fluorescence, ultrastructural, silver-impregnation and immunohistochemical methods in the same section or in consecutive semi-thin and ultra-thin sections. The results indicate that islet cells emitting intense FIF exhibit a strongly argyrophil reaction with the Grimelius' silver method and also immunohistochemical reaction with anti-glucagon serum, but not with anti-5-HT serum. Therefore, the fowl islet A cell, a peptide hormone-producing cell, stores simultaneously catecholamine as biogenic amine. The islet B and D cells did not display any FIF, any argyrophil reaction with the Grimelius' silver method, or any immunoreactivity with anti-glucagon or anti-5-HT sera. The fluorescent but non-argyrophil cells dispersed in the exocrine acinus may well be PP cells.
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  • 36
    ISSN: 1432-0878
    Keywords: Pineal complex (pineal and parapineal organs) ; Development, ontogenetic ; Photoreceptor cells ; Immunohistochemistry ; Serotonin (5-HT) ; Opsin ; Teleost (Gasterosteus aculeatus)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 5-hydroxytryptamine (5-HT, serotonin)- and opsin-immunoreactive sites were studied in the developing pineal complex of the stickleback, Gasterosteus aculeatus L., by use of light-microscopic indirect immunoperoxidase techniques. 5-HT immunoreactivity first occurs in the pineal organ at the age of 80 h after fertilization and appears to be localized in cells of the photoreceptor type. The outer segments of a few pineal photosensory cells exhibit opsin immunoreactivity at the age of 84 h after fertilization. The number of cells seems to increase until the pineal organ is completely developed. The increase in the number of 5-HT immunoreactive perikarya runs parallel in time to that of the opsinimmunoreactive outer segments. The cells of the parapineal organ show neither opsin nor 5-HT immunoreactivity. The retina of the embryonic stickleback does not display opsin immunoreactivity until after hatching, which takes place about 144 h after fertilization. These results suggest, in the three-spined stickleback, an earlier light-perception capacity for the developing pineal organ than for the retina.
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  • 37
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    Cell & tissue research 238 (1984), S. 421-423 
    ISSN: 1432-0878
    Keywords: Gut endocrine cells ; Ontogeny ; Immunohistochemistry ; Pancreatic polypeptide ; Insect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ontogeny of the endocrine cells of the gut of the cockroach Periplaneta americana was studied by immunohistochemistry. During embryogenesis, the midgut begins to be formed as an outgrowth of the foregut and hindgut invaginations. Gut endocrine cells with pancreatic polypeptide (PP)-like immunoreactivity begin to appear at the anterior and posterior ends of the forming midgut. These cells are restricted to the midgut epithelium, and no mitotic cells with PP-like immunoreactivity are observed. These results strongly suggest that the gut endocrine cells, at least those with PP-like immunoreactivity, are derived from precursor cells they have in common with other epithelial cells of the midgut.
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  • 38
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    Cell & tissue research 259 (1990), S. 93-103 
    ISSN: 1432-0878
    Keywords: Metamorphosis ; Imaginal disc ; Sensory neurons ; Immunohistochemistry ; Phormia regina, Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The development of the sensory neurons of the legs of the blowfly,Phormia regina has been described from the third instar larva to the late pupa using immunohistochemical staining. The leg discs of the third instar larva contain 8 neurons of which 5 come to lie in the fifth tarsomere of the developing leg. Whereas 2 neurons persist at least to the late pupa, the other cells degenerate. The first neurons of gustatory sensilla arise in the fifth tarsomere at about 1.5 h after formation of the puparium. Most of these sensilla, however, appear within a short time period beginning at about 18 h. The femoral chordotonal sensory neurons first appear at the time of formation of the puparium, as a mass of cells situated in the distal femur. During later pupal development 2 groups of these cells come to lie at the femur-trochanter border, where they become the proximal femoral chordotonal organ of the adult; the remaining cells become the distal femoral chordotonal organ. Other scolopidial neurons appear later in development. The nerve pathways of the late pupal leg are established either by the axons of the cells that are present in the larval leg disc or by new outgrowing processes of sensory neurons. In the tibia, the initial direction of new outgrowth differs in different regions of the segment: proximal tibial neurons grow distally, while distal tibial neurons grow initially proximally.
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  • 39
    ISSN: 1432-0878
    Keywords: Vasopressin ; Oxytocin ; Tyrosine hydroxylase ; Dopamine ; Galanin ; Dynorphin ; Cholecystokinin ; Salt-loading ; Supraoptic nucleus ; Paraventricular nucleus ; Neurophypophysis ; In situ hybridization ; Immunohistochemistry ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In situ hybridization histochemistry and indirect immunofluorescence histochemistry were used to study changes in the expression of vasopressin (VP), oxytocin (OXY), tyrosine hydroxylase (TH), galanin (GAL), dynorphin (DYN) and cholecystokinin (CCK) in hypothalamic magnocellular neurons of the paraventricular (PVN) and supraoptic (SON) nuclei of rats. After prolonged administration of 2% sodium chloride as drinking water (salt-loading), the treatment increased the levels of VP, OXY, TH, GAL, DYN and CCK mRNA in the PVN and SON. The increase in CCK mRNA was, however, proportionally higher in the PVN than in the SON. Within cell bodies of the PVN and SON of salt-loaded rats, a depletion of VP- and OXY-like immunoreactivity (LI) and an increase in TH-LI were seen. In salt-loaded/colchicine-treated rats, a marked decrease in GAL- and DYN-LI, but no specific changes in CCK-LI were observed. Within nerve fibers of the posterior pituitary of salt-loaded rats, a marked depletion of VP-, GAL- and DYN-LI was found. Less pronounced depletion was observed in OXY- and CCK-LI, and no specific changes in TH-LI were seen. The results show that high plasma osmolality induces increased mRNA levels for VP, OXY, TH, GAL, DYN and CCK, presumably indicating increased synthesis, an increased export from cell somata of VP, OXY, GAL and DYN, and a decrease in levels of these peptides in the posterior pituitary, suggesting increased release. The catecholamine-synthesizing enzyme TH, however, which has a cytoplasmic localization and is not released from nerve endings, remains high in the cell bodies and nerve endings during this state of increased activity.
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  • 40
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    Cell & tissue research 260 (1990), S. 415-419 
    ISSN: 1432-0878
    Keywords: GABA ; Vestibular organ ; Hair cells ; Immunohistochemistry ; Guinea pig (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Neurotransmitters involved in the vestibular system are largely uncharacterized. On the basis of results of earlier electrophysiological and immunohistochemical experiments, glutamate and gamma-amino-butyric acid (GABA) have been proposed in both mammalian and non-mammalian species as afferent transmitters between the sensory cell and the afferent dendrite. GABA is also suspected to act as an efferent neurotransmitter in the cochlea. We describe in this study the immunocytochemical localization of GABA within the vestibular end organs in the guinea pig. GABA immunoreactivity was found in the calyceal nerve endings surrounding type I hair cells of the vestibular epithelia. The most significant labelings were obtained in the crista ampullaris. Labeling was more difficult to observe in the utricular and saccular macula. These results contribute to the recent proposal that the calyx has a secretory function, and suggest that GABA may have a modulatory influence upon the type I hair cells.
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  • 41
    ISSN: 1432-0878
    Keywords: Substance P ; Neuropeptide Y ; Calcitonin gene-related peptide (CGRP) ; Cholecystokinin (CCK) ; Immunohistochemistry ; Neuropeptide coexistence ; Planorbarius corneus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of the vertebrate-like neuropeptides substance P (SP), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and cholecystokinin (CCK8) in the central nervous system of the freshwater snail Planorbarius corneus has been studied using specific antisera and single and double immunohistochemistry. A widespread but precise distribution of immunore-activity (IR) in neurons and fibers of almost all the ganglia is observed for each antiserum. A comparison of the IR with classical neurosecretory staining (AB/AY) shows a partial overlap only for CGRP and CCK8. Whereas CGRP-IR is found in some Yellow Cells in the left parietal ganglion, CCK8-IR is found in Yellow Green, Green and Brown Cells in the viscero-parietal complex. Studies employing double-sequential methods or simultaneous immunofluorescence have shown that, with regard to the tested antisera, CCK8- and NPY-IR are colocalized in a limited number of cells and fibers in the buccal and visceral ganglia, whereas CCK8- and SP-IR are colocalized only rarely in neurons in the left cerebral ganglion. The possible roles in P. corneus of the investigated neuropeptides and the contribution that molluscan models may offer to the knowledge of the basic properties of neuropeptides are discussed.
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  • 42
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    Cell & tissue research 261 (1990), S. 323-331 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Innervation ; Substance P ; Immunohistochemistry ; Dog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pars distalis of the anterior pituitary is known to be regulated by hypothalamic hormones. Recently, we have discovered the presence of substance P-like immunoreactive nerve fibers in the pars distalis of the monkeys. Substance P-like immunoreactivity in the pars distalis of the dog was investigated in this study. A substantial amount of substance P-like immunoreactive nerve fibers with a large amount of varicosities were found. They were widely distributed in the gland, more abundant along its periphery. Most of them were closely related to the glandular tissue, some were located on vascular walls. Substance P-like immunoreactive nerve fibers were also found in the meningeal sheath of the anterior pituitary. They could be followed into the parenchyma of the gland.
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  • 43
    ISSN: 1432-0878
    Keywords: Gut ; Macrophages ; High-endothelial venules ; Immunohistochemistry ; Mouse BALB/c
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary This study concerns the distribution of macrophages and dendritic cells (DC) in the gastrointestinal tract of the mouse. Heterogeneity of macrophage population was found by using the MOMA-1, MOMA-2, ERTR-9, Mac-1 and F4/80 monoclonal antibodies. MOMA-1, ERTR-9, Mac-1 and F4/80+ cells were detected mostly at the villous cores in the lamina propria of the villi, whereas MOMA-2+ cells were primarily found around the crypts at the base of the villi. These MOMA-2+ cells revealed a granular appearance throughout the cytoplasm and displayed a strong acid phosphatase (AcPh) activity. Few MOMA-2+ cells were seen at the top of the villi in the epithelium. Although MOMA-1 and ERTR-9+ cells have similar morphology and the same distribution patterns in the lamina propria, they are likely different populations, because in Peyer's patches (PP), MOMA-1+ cells were present, whereas ERTR-9+ cells could not be detected. Both populations displayed AcPh activity. Strongly stained Mac-1+ cells were abundantly seen in the lamina propria of the small intestine. F4/80+ cells were rare. NLDC-145+ cells with AcPh activity and weak Ia staining were also found. In the PP-associated villi and in the T-dependent area of PP, dendritic NLDC-145+ cells, which were strongly Ia positive, were detected. MIDC-8+ cells were found only in the T-dependent area. Few NLDC-145+ cells (dendritic cells) were found in the upper part of the oesophagus. These cells were also stained with the MIDC-8 antibody. The MECA-325 monoclonal antibody recognized high endothelial venules (HEV) in PP and blood vessels at the base of the villi of the jejunumileum and caecum. Unlike in PP, the endothelium of the venules in the villi was flat.
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  • 44
    ISSN: 1432-0878
    Keywords: Met-enkephalin ; Immunohistochemistry ; Intestine, small ; Neuron, enteric ; Laser microsurgery ; Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve elements in the small intestine of the bullfrog. Rana catesbeiana, were studied by immunohistochemistry with anti-methionine enkephalin antisera and by nerve lesion experiments, using laser irradiation. Methionine-enkephalin immunopositive nerve fibers occur in the myenteric plexus, circular muscle layer, submucosa, and mucosa. Immunopositive nerve cell bodies in the myenteric plexus have dendrite-like and a long axon-like processes. In the froglet (3 months after metamorphosis), these axon-like processes lead posteriorly in the nerve strand of the myenteric plexus. Some bifurcate, one branch continuing posteriorly, the other doubling back to lead anteriorly; both form terminal varicose fibers in the circular muscle layer. Nerve lesion experiments, in the adult bullfrog, resulted in accumulations of methionine-enkephalin immunoreactivity at the oral and hinder edges of the laser-irradiated necrotic area; there were sprouting and nonsprouting immunopositive stumps. It is suggested that bidirectional flow of methionine-enkephalin in the myenteric plexus is mediated via the anterior and posterior branches of the axon-like process. The difference in sprouting behavior of immunopositive nerve fiber stumps, after nerve lesion, is discussed with reference to regional differences of the axon-like process.
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  • 45
    ISSN: 1432-0878
    Keywords: Cholera toxin B subunit ; Motoneurons ; Primary afferent neurons ; Spinal cord ; Retrograde transport ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Morphological changes in the motor and sensory neurons in the lumbar spinal cord and the dorsal root ganglia were investigated at different survival times following the injection of the B subunit of cholera toxin (CTB) into the medial gastrocnemius muscle. Unconjugated CTB, visualized immunohistochemically, was found to be retrogradely transported through ventral and dorsal roots to motor neurons in the anterior horn, each lamina in the posterior horn, and ganglion cells in the dorsal root ganglia at L3–L6. The largest numbers of labeled motor neurons and ganglion cells were observed 72 h after the injection of CTB. Thereafter, labeled ganglion cells were significantly decreased in number, whereas the amount of labeled motor neurons showed a slight reduction. Motor neurons had extensive dendritic trees filled with CTB, reaching lamina VII and even the pia mater of the lateral funiculus. Labeling was also seen in the posterior horn, but the central and medial parts of laminae II and III had the most extensively labeled varicose fibers, the origin of which was the dorsal root ganglion cells. The results indicate that CTB is taken up by nerve terminals and can serve as a sensitive retrogradely transported marker for identifying neurons that innervate a specific muscle.
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  • 46
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    Cell & tissue research 272 (1993), S. 147-154 
    ISSN: 1432-0878
    Keywords: Histamine ; Immunohistochemistry ; Brain, vertebrate ; Catecholamines ; Triturus carnifex (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of immunoreactivity for histamine was studied in the brain of the urodele Triturus carnifex using the indirect immunofluorescence method. Histamine-immunoreactive cell bodies were localized in the caudal hypothalamus within the dorsolateral walls of the infundibular recesses. These immunoreactive cell bodies were pear-shaped, bipolar and frequently of the cerebrospinal-fluid-contacting type. Histaminergic nerve fibers were detected in almost all parts of the brain. Dense innervation was seen in the telencephalic medial pallium and ventral striatum, the neuropil of the preoptic area, the septum, the paraventricular organ, the posterior commissure, the caudal hypothalamus, the ventral and lateral mesencephalic tegmentum. Medium density innervation was observed in the lateral mesencephalic tegmentum and optic tectum. Poor innervation was present in the telencephalic dorsal pallium and in the central gray of the medulla oblongata. Few fibers occurred in the olfactory bulbs and in the telencephalic lateral pallium. Double immunofluorescence staining, using an antibody against tyrosine hydroxylase, showed that histamine-immunostained somata and those containing tyrosine-hydroxylase-like immunoreactivity were co-distributed in the tuberal hypothalamus. No co-occurrence of histamine-like and tyrosine hydroxylase-like immunostaining was seen in the same neuron. The pattern of histamine-immunoreactive neurons in the newt was similar to that described in other vertebrates. Our observations, carried out on the apparently simplified brain of the newt confirm that the basic histaminergic system is well conserved throughout vertebrates.
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  • 47
    ISSN: 1432-0878
    Keywords: Serotonin ; Pancreas ; Phylogenic study ; Immunohistochemistry ; Teleosts ; Chicken ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution pattern of serotonin (5HT) in the pancreas was studied immunohistochemically by using a 5HT monoclonal antibody in various vertebrates including the eel, bullfrog, South African clawed toad, turtle, chicken, mouse, rat, guinea-pig, cat, dog and human. In all species examined, except the bullfrog, 5HT-like immunoreactivity was observed in nerve fibers, in endocrine cells, or in both. Positive nerve fibers were found in the eel, turtle, mouse, rat and guinea-pig. These fibers ran mainly along the blood vessels and partly through the gap between the exocrine glands. In the eel and guinea-pig, positive fibers invaded the pancreatic islet. Occasionally, these positive fibers were found adjacent to the surface of both exocrine and endocrine cells, suggesting a regulatory role of 5HT in pancreatic function. 5HT-positive endocrine cells were observed in the pancreas of all species except for the bullfrog and rat. In the eel and in mammals such as the mouse, guinea-pig, cat, dog and human, 5HT-positive cells were mainly observed within the pancreatic islet. In the South African clawed toad, turtle and chicken, the positive cells were mainly in the exocrine region. The present study indicates that the distribution patterns of 5HT in the pancreas varies considerably among different species.
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  • 48
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    Cell & tissue research 263 (1991), S. 431-438 
    ISSN: 1432-0878
    Keywords: Connective tissue ; EHS tumor ; Basement membrane ; Amyloid P component ; Immunohistochemistry ; Mouse (C57BL/6)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A new minute connective tissue structure, referred to as “pentosome”, has been investigated by electron microscopy and its nature has been examined by immunoperoxidase tests. Pentosomes are 3.5-nm wide, particulate structures that have been observed in the posterior chamber of the eye, the connective tissue spaces of the mouse foot-pad and the matrix of the mouse EHS tumor. They are usually found in the vicinity of microfibrils whether they are free or associated with elastic fibers. They tend to be organized into groups forming a three-dimensional semi-crystalline lattice at 10-nm intervals, but are connected by fine filaments. At high magnification, pentosomes appear as hollow structures composed of two parallel pentagons, which respectively measure 2.7 and 3.5 nm, and are held together by a cross-bar. A series of immunoperoxidase tests has only shown antigenicity against a serum protein, the amyloid P component. However, pentosomes are only about one-third the size of the 8.5-nm wide, disk-like segments of the amyloid P molecule. Since they could be subunits of these molecules, such subunits were prepared and compared with pentosomes; they appeared to be identical. It is concluded that the pentosomes found in connective tissue are AP subunits.
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  • 49
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    Cell & tissue research 268 (1992), S. 341-358 
    ISSN: 1432-0878
    Keywords: Neuronal development ; Neuropil ; Immunohistochemistry ; Serotonin ; FMRF-amide ; Brain, invertebrate ; Protocerebrum ; Tenebrio molitor (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuroarchitecture of the central complex, a prominent neuropil in the midbrain of the holometabolan, Tenebrio molitor, is described throughout larval development. The analysis is based on classical silver impregnations and on fate-mapping of identified neurons using antisera against serotonin and FMRF-amide. In T. molitor, the central body is present in the first larval instar, and is formed by side branches of contralaterally projecting neurons. Glial cells surround eight neuropil compartments in the first larval instar. These subdivisions in the organization of the fan-shaped body are maintained throughout development. Intrinsic interneurons are found from the 5th larval instar onwards. In the last larval stage, the central complex consists of the fan-shaped body, the protocerebral bridge, and the anlage of the ellipsoid body. The cellular architecture of the fan-shaped body of the last larval instar resembles the basic structural characteristics of the adult. Serotonin-immunoreactive neurons and FMRF-amide immunoreactive neurons in the midbrain of the first larval instar show the basic structural features of the respective imaginal cells. The structural organizations of larval and adult midbrain are compared.
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  • 50
    ISSN: 1432-0878
    Keywords: Tyrosine hydroxylase ; Dopamine ; Serotonin ; Immunohistochemistry ; CNS amphibian ; Triturus alpestris (Urodela)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Mapping of monoaminergic systems in the brain of the newt Triturus alpestris was achieved with antisera against (1) thyrosine hydroxylase (TH), (2) formaldehyde-conjugated dopamine (DA), and (3) formaldehyde-conjugated serotonin (5-HT). In the telencephalon, the striatum was densely innervated by a large number of 5-HT-, DA-and TH-immunoreactive (IR) fibers; IR fibers were more scattered in the amygdala, the medial and lateral forebrain bundles, and the anterior commissure. In the anterior and medial diencephalon, TH-IR perikarya contacting the cerebrospinal fluid (CSF-C perikarya) were located in the preoptic recess organ (PRO), the organum vasculosum laminae terminalis and the suprachiasmatic nucleus. Numerous TH-IR perikarya, not contacting the CSF, were present in the posterior preoptic nucleus and the ventral thalamus. At this level, DA-IR CSF-C neurons were only located in the PRO. In the posterior diencephalon, large populations of 5-HT-IR and DA-IR CSF-C perikarya were found in the paraventricular organ (PVO) and the nucleus infundibularis dorsalis (NID); the dorsal part of the NID additionally presented TH-IR CSF-C perikarya. Most regions of the diencephalon showed an intense monoaminergic innervation. In addition, numerous TH-IR, DA-IR and 5-HT-IR fibers, orginating from the anterior and posterior hypothalamic nuclei, extended ventrally and reached the median eminence and the pars intermedia of the pituitary gland. In the midbrain, TH-IR perikarya were located dorsally in the pretectal area. Ventrally, a large group of TH-IR cell bodies and some weakly stained DA-IR and 5-HT-IR neurons were observed in the posterior tuberculum. No dopaminergic system equivalent to the substantia nigra was revealed. The possible significance of the differences in the distribution of TH-IR and DA-IR neurons is discussed, with special reference to the CSF-C neurons.
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  • 51
    ISSN: 1432-0878
    Keywords: Neural development ; Plasticity ; Serotonin ; Immunohistochemistry ; Coho salmon, Oncorhynchus kisutch (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In coho salmon (Oncorhynchus kisutch), smolt transformation has been shown to be associated with sequential surges of neurotransmitters in the brain. In order to determine if the surge of serotonin (5-HT) is correlated with structural changes, we have used immunocytochemistry to observe changes in the serotonin immunoreactivity before, during and after the 5-HT surge. The following stages were studied: 12-month-old freshwater presmolts, 17-month-old freshwater presmolts, 18-month-old saltwater smolts, 19-month-old saltwater postsmolt, 24-month-old postsmolt, and adult spawners. In the 17-month-old samples, but not at any other stage, we found a set of transient (serotonin-immunoreactive) 5-HT-immunoreactive neurons in the lateral preoptic area, as well as a discrete population of 5-HT-immunoreactive neurons in the lateral part of the dorsal right habenular nucleus. In addition, a higher density of serotonergic fibers was found in the telencephalon at this stage compared to the following two stages. Since the transient 5-HT-immunoreactive structures presented here do not appear simultaneously with the 5-HT total brain concentration surge, we conclude that they are unlikely to be the source of the 5-HT surge, but are probably related to other developmental changes in the brain associated with smolt transformation.
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  • 52
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    Cell & tissue research 268 (1992), S. 513-519 
    ISSN: 1432-0878
    Keywords: Placenta ; Macrophages ; Reticulum cells ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The placenta is a unique mixture of histoincompatible cells derived from mother and fetus. The aim of the present study was to obtain information on the development of macrophage subpopulations and reticulum cells during pregnancy in the placenta. Placentas of Wistar rats were removed at several stages of gestation, and were studied by immunohistochemical techniques applying monoclonal antibodies against macrophage subpopulations, lymphoid cells and reticulum cells. The expression of MHC class-II antigens was also studied. Throughout gestation macrophages were demonstrable in large numbers in the endometrium, in the myometrium and in the metrial gland, which is a compartment developing in the myometrium of pregnant rodents. In the labyrinth, a placenta compartment consisting of fetal cells, macrophages (probably of fetal origin) were already found on day 15. In the spongiotrophoblast and decidua basalis, which are layers of the placenta containing both maternal and fetal cells, only a few macrophages were recognized throughout gestation. The monoclonal antibody ED11, raised against reticulum cells, recognized fiber-like structures lining the blood sinuses of the spongiotrophoblast, in which only maternal blood is circulating. As the antigen recognized by ED11 is believed to play a role in the trapping of immune complexes, the spongiotrophoblast may play a role in the protection of the fetus from circulating immune complexes.
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  • 53
    ISSN: 1432-0878
    Keywords: FMRFamide ; Catch-relaxing peptide (CARP) ; Radula muscle ; Immunohistochemistry ; Rapana thomasiana (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In a prosobranch mollusc, Rapana thomasiana, the catch-relaxing peptide H-Ala-Met-Pro-Met-Leu-Arg-Leu-NH2 (CARP) was found to depress the contraction of the radula protractor and retractor elicited by electrical stimulations. The action of CARP was in contrast to that of other neuropeptides, H-Phe-Met-Arg-Phe-NH2 (FMRFamide) and H-Phe-Leu-Arg-Phe-NH2 (FLRFamide), which enhanced the contraction of the radula protractor and retractor, respectively. By immunohistochemical examinations, FMRFamide-like immunoreactive neurons were found on the rostral side of the right buccal ganglion and the caudal side of the left ganglion, where some CARP-like immunoreactive neurons were also distributed, indicating a possible coexistence of FMRFamide and CARP. FMRFamide- and CARP-like immunoreactivities were also detected in the neuropile of buccal ganglia, radula nerves arising from the ganglia, and nerve fibers in the radula muscles. The present results suggest that FMRFamide- and CARP-like peptides are involved in the regulation of the contraction of the radula muscles.
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  • 54
    ISSN: 1432-0878
    Keywords: Bromodeoxyuridine ; Immunohistochemistry ; Fundic gland ; Stomach ; Development ; Xenopus laevis (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cellular differentiation and migration in the fundic glands of adult and larval Xenopus laevis have been examined using bromodeoxyuridine-immunohistochemistry. In the adult fundic gland, cumulative labeling with bromodeoxyuridine revealed a proliferative cell zone between the surface mucous cells and mucous neck cells, in what is referred to as the neck portion of the gland. The labeling-index of mucous neck cells had rapidly increased by week-5. The labeling-index of oxynticopeptic cells showed a more delayed increase until week-7, coincident with the decrease in the labeling of mucous neck cells. In the immature fundic glands of larvae, the labeled proliferating cells were randomly distributed throughout the developing gastric mucosa. During metamorphosis, the labeling-index of immature epithelial cells was highest at stage 63. Following administration of bromodeoxyurdine at this, stage, there was no significant loss of labeled epithelial cells during the metamorphosing period. Furthermore, there was no significant difference in the labeling-indices among the epithelial cells, such as surface mucous cells/generative cells, mucous neck cells, and oxynticopeptic cells, 7 days after administration. Cellular differentiation and migration pathways of epithelial cells in the fundic gland of adult X. laevis and its larvae are discussed.
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  • 55
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    Cell & tissue research 269 (1992), S. 289-297 
    ISSN: 1432-0878
    Keywords: Calbindin D-28k ; Olfactory bulb ; Calcium-binding proteins ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have studied the distribution of calbindin D-28k immunoreactivity in the rat olfactory bulb using specific monoclonal antibodies and the avidin-biotin-immunoperoxidase method. The largest number of positive neurons was located in the periglomerular layer. These neurons were identified as periglomerular cells; they have been described also by other authors as calbindin-positive elements. Close to these neurons, a second population of nerve cells was identified as superficial shortaxon neurons. The remaining layers showed a smaller number of stained elements. Other labeled neurons were located along the external border of the external plexiform layer; the scarce neurons marking its internal border were identified as van Gehuchten cells. No immunoreactive structures were found in the mitral cell layer, although we observed another population of immunostained short-axon cells at its internal border. Some reactive structures, identified by us as horizontal and vertical cells of Cajal, were located in the boundary zone between the internal plexiform layer and the granule layer. In the white matter, we found a neuronal type characterized by its large size and oriented arborization of varicose dendrites.
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  • 56
    ISSN: 1432-0878
    Keywords: α2-Macroglobulin receptor ; Low-density lipoprotein receptor-related protein ; Tissue distribution ; Brain ; Macrophages ; Immunohistochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The hepatic α1-macroglobulin receptor (α2MR)/low density lipoprotein receptor-related protein (LRP) binds and endocytoses α2-macroglobulin-proteinase complexes in plasma. In addition, it binds lipoproteins, a novel 40 kDa protein, and complexes between plasminogen activators and plasminogen activator inhibitor type-1. This study shows, for the first time, the tissue distribution of α2MR/LRP as determined by immunohistochemistry with specific monoclonal antibodies. The analysis revealed α2MR/LRP-expression in a restricted spectrum of cell types, including neurons and astrocytes in the central nervous system, epithelial cells of the gastrointestinal tract, smooth muscle cells, fibroblasts, Leydig cells in testis, granulosa cells in ovary, and dendritic interstitial cells of kidney. Monocytederived cells displayed marked α2MR/LRP expression in the phagocytes of liver, lung and lymphoid tissues, but no or low expression in antigen-presenting cells including Langerhans' cells of the skin. The high abundance of α2MR/LRP in certain cell types of most organs suggests two main routes for α2MR/LRP ligand clearance: (1) systemic removal in liver of circulating ligands, and (2) non-hepatic interstitial removal in different organs, including the brain.
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  • 57
    ISSN: 1432-0878
    Keywords: Peyer's patches ; Collagen ; Laminin ; Fibronectin ; Extracellular matrix ; Lymphocytes ; Immunohistochemistry ; Mouse (BALB/c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution patterns of extracellular matrix elements were determined to ascertain whether they play a role in the localization of lymphocytes in discrete T-cell, B-cell and dome antigen-processing domains within Peyer's patches. Antibodies against collagen types I, III and IV, laminin and fibronectin were applied to cryosections of mouse Peyer's patches and localized by direct or indirect immunoperoxidase methods. T-cell domains were identified with a monoclonal antibody against Thy-1.2. Labeled reticular fibers in distinctive patterns were more numerous in parafollicular and dome areas than within follicles. Germinal centers contained few such fibers. In parafollicular areas, fibers were oriented predominantly toward follicle domes; their distribution corresponded to T-cell zones and lymphocyte traffic areas, with their orientation being parallel to the migration pathways of lymphocytes from high endothelial venules to the antigen-processing domes. Subepithelial and subendothelial basal laminae were immunopositive for type-IV collagen, laminin and fibronectin. The dome subepithelial basal lamina had pore-like discontinuities through which lymphocytes migrated to and from the epithelium. The correspondence of the distribution patterns of extracellular matrix to specific functional domains of Peyer's patches suggests that this matrix provides a structural framework for lymphocyte migration and localization.
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  • 58
    ISSN: 1432-0878
    Keywords: Immunohistochemistry ; Pelvic nerves ; Neuropeptides ; Large intestine ; Bufo marinus (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution and colocalization of neuropeptides and 5-hydroxytryptamine in the posterior portion of the large intestine of the toad was studied using single- and dual-label immunohistochemistry. Neurons containing colocalized galanin/somatostatin or vasoactive intestinal peptide alone were observed along intramural pelvic nerves. Some of the galanin/somatostatin neurons also contained 5-hydroxytryptamine. Synaptic boutons containing colocalized calcitonin gene-related peptide/vasoactive intestinal peptide were associated with the galanin/somatostatin neurons. The muscle of the large intestine was also innervated by axons containing galamin/somatostatin, vasoactive intestinal peptide/calcitonin gene-related peptide or vasoactive intestinal peptide alone. Nerve fibres containing calcitonin gene-related peptide/substance P, probably representing primary afferent nerves, were also associated with muscle bundles. Submucosal blood vessels carried dense plexuses of fibres containing vasoactive intestinal peptide alone or and calcitonin gene-related peptide/substance P. Adrenergic perivascular nerves also contained galanin and neuropeptide Y.
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  • 59
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    Cell & tissue research 269 (1992), S. 525-534 
    ISSN: 1432-0878
    Keywords: Calcitonin gene-related peptide (CGRP) ; Immunohistochemistry ; Nervous system, central ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of calcitonin gene-related peptide (CGRP), immunoreactive structures in the central nervous system of the frog, Rana esculenta, was studied using the peroxidase immunohistochemical method. Immunoreactive perikarya were found in all major parts of the brain. In the forebrain, neurons of the septohipocampal formation, the amygdala, the ventromedial and posterocentral thalamic nuclei, and the cerebrospinal fluid contacting neurons in the diencephalic periventricular organ showed immunoreactivity. The pear-shaped neurons of the optic tectum, and perikarya of the oculomotor nucleus in the midbrain were also immunoreactive. In the hindbrain, neurons of the cranial nerve motor nuclei, neurons of the superior vestibular nucleus, giant cells of the reticular formation, and preganglionic parasympathetic neurons of the superior salivatory nucleus were stained. Motoneurons presented immunostaining also in the spinal cord. Immunoreactive fibers were shown to occur in the olfactory tract, the striatum, the tegmentum and the basis mesencephali, the descending tract of the trigeminal nerve, the solitary tract, Lissauer's tract, and the dorsal horn of spinal cord. A comparison of the distribution of CGRP immunoreactivity in the mammalian and amphibian central nervous system revealed that, in relation to the size of the brain, CGRP is more extensively distributed in the amphibian than in the mammalian limbic system.
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  • 60
    ISSN: 1432-0878
    Keywords: Calbindin ; Immunohistochemistry ; Teleosts ; Visual system ; Entreleation ; Cyprinus carpio ; Tinca tinca (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the ABC immunohistochemical method, we investigated the distribution of calbindinlike immunoreactive structures in the optic tectum of normal fish, Tinca tinca, and from normal and unilaterally eye-enucleated fish, Cyprinus carpio. In nonoperated individuals of both species the optic tectum contained numerous immunoreactive neurons with strongly positive somata located in the stratum periventriculare and a thick immunolabeled dendritic shaft ascending radially toward the stratum fibrosum et griseum superficiale. The retinorecipient layers contained many fibrous immunoreactive structures. Some varicose fibers, isolated or in small bundles, were localized to the stratum album centrale, especially in the dorsal tectal half. Unilateral eye removal produced the disappearance of the immunoreactive fibrous structures located in the retinorecipient layers of the tectum contralateral to the enucleation. The present work shows that calbindinlike immunoreactive substances are localized in specific neural circuits of the fish optic tectum and suggests that the calbindin-like immunoreactive fibers in the retinorecipient strata are of retinal origin.
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  • 61
    ISSN: 1432-0878
    Keywords: L2/HNK-1 carbohydrate epitope ; L3 carbohydrate epitope ; Immunohistochemistry ; Extracellular matrix ; Calliphora vicina (Insecta) ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The carbohydrate epitopes L2/HNK-1 and L3 belong to two overlapping families of adhesion molecules in the vertebrate, and probably the invertebrate nervous systems. To investigate their pattern of expression during the development of insects, cryosections of late third instar larvae and imagoes of Drosophila melanogaster and Calliphora vicina were studied by indirect immunofluorescence using several monoclonal antibodies to the L2/HNK-1 and one monoclonal antibody to the L3 epitope. Each monoclonal antibody to the L2/HNK-1 epitope showed a different immunohistological staining pattern, which differed from that of the L3 monoclonal antibody. In both insect species the immunohistological staining patterns for the two carbohydrate epitopes were similar at the two developmental stages, with immunoreactivity not confined to the nervous system. In larvae, immunoreactivities of the monoclonal antibodies L2.334 and L3.492 were predominantly associated with the extracellular matrix as indicated by co-localization with laminin, particularly in the imaginal discs, while L2.349 revealed a more cell surface-associated distribution. In imagoes, immunoreactivities were detectable in most organs studied.
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  • 62
    ISSN: 1432-0878
    Keywords: FMRFamide ; ACEP-1 (Achatina cardio-excitatory peptide-1) ; Cardiac regulation ; Immunohistochemistry ; Achatina fulica (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical localization of two neuropeptides possibly involved in the regulation of cardiac activity in a pulmonate mollusc, Achatina fulica Férussac, was studied. On the ventral surface of the right cerebral ganglion, more than 50 neurons with diameters of 30–50 μm showed immunoreactivity to the antiserum of the neuropeptide FMRFamide. Many were also immunoreactive to an antiserum raised against Achatina cardio-excitatory peptide-1 (ACEP-1). Although FMRFamidelike immunoreactive neurons occurred in all components of the subesophageal ganglia, identifiable ACEP-1-like immunoreactive neurons were located only in the visceral ganglion and the right parietal ganglion. In the heart, FMRFamide- and ACEP-1-like immunoreactive fibers were restricted to the atrium and the aortic end of the ventricle, consistent with morphological observations of cardiac innervation. The present results suggest that FMRFamide-and ACEP-1-like peptides are involved in regulating the heart beat of this snail.
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  • 63
    ISSN: 1432-0878
    Keywords: Neurotoxins ; Immunohistochemistry ; Macrophages ; Dendritic reticulum cell ; B-cells ; Indoleamines ; NADPH oxidase ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Antibodies to quinolinic acid were produced in rabbits with protein-conjugated and gold particle-adsorbed quinolinic acid. Quinolinic acid immunoreactivity was below detection limits in carbodiimide-fixed rat brain. In contrast, strong quinolinic acid immunoreactivity was observed in spleen cells with variable, complex morphology located predominantly in the periarterial lymphocyte sheaths. In the thymus, quinolinic acid immunoreactivity was observed in cells with variable morphology, located almost exclusively in the medulla. Lymph nodes and gut-associated lymphoid tissue contained many, strongly stained cells of similar complex morphology in perifollicular areas. Immunoreactivity in liver and lung was restricted to widely scattered, perivascular cells and alveolar cells respectively. Additional stained cells with complex morphology were observed in bronchus-associated lymphoid tissue, in skin, and in the lamina propria of intestinal villi. Follicles in all secondary lymphoid organs were diffusely stained, ranging from mildly to moderately immunoreactive in spleen, to intensely immunoreactive in gut-associated lymphoid tissue. These results suggest that quinolinic acid is an immune system-specific molecule. Two hypothetical schemes are proposed to account for high levels of quinolinic acid in specific cells of the immune system.
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  • 64
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    Cell & tissue research 278 (1994), S. 573-578 
    ISSN: 1432-0878
    Keywords: Key words: Placenta ; Giant cells ; Chorionic gonadotropin ; Luteotropin ; Electrophoresis ; Immunohistochemistry ; Rabbit
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    Notes: Abstract. Obplacental giant cells are enlarged cells, found following implantation, in the antimesometrial region of the rabbit uterus. They probably originate from trophoblastic knobs that traverse the uterine epithelium during early implantation. Little is known about their function. In this study, trophoblast, placental, paraplacental and obplacental tissues at days 7–15 post-coitum, and enzyme-isolated giant cells at day 15 were studied by two-dimensional gel electrophoresis, followed by immunoblotting and light-microscopic immunohistochemistry, for the presence of human chorionic gonado- tropin-like proteins. Immunostaining was performed by using anti-human chorionic gonadotropin antibodies. In gel electrophoresis of obplacental tissue and isolated giant cells, two proteins of human chorionic gonadotropin-like antigenicity at 26 kDa with pIs equivalent to pH 6.4 and 6.6 were found; they were absent in the placenta, paraplacenta, day-7 blastocyst and day-8 trophoblast. The onset of synthesis of these proteins could be observed when day-8 trophoblastic tissue was cultured in vitro for 24 h. In immunohistochemistry, only the obplacental giant cells showed a positive reaction, indicating that the production of chorionic gonadotropin occurs in this cell type.
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  • 65
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    Keywords: Key words: FSH ; Immunohistochemistry ; Receptor mRNA ; In situ hybridization ; Sertoli cell ; Testis ; Human
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    Notes: Abstract. Testicular biopsies from 82 oligo- or azoospermic male patients were subjected to immunostaining using anti-human FSH antibodies. Histological evaluation showed normal spermatogenesis (nspg) in 7 (FSH: 2.7±0.7), mixed atrophy (ma) in 63 (FSH: 5.3±0.5), and bilateral or unilateral Sertoli Cell Only syndrome (SCO) in 12 (FSH:21.7±3.5) patients. For the relationship between FSH values and testicular histology, see Bergmann et al. (1994). FSH immunoreactivity was found exclusively in Sertoli cells and in some interstitial cells. Seminiferous epithelium showing normal or impaired spermatogenesis displayed only weak immunoreactivity compared to intense immunoreaction, i.e. large and numerous vesicles in Sertoli cells of SCO tubules in biopsies showing mixed atrophy or SCO. In addition, h-FSH receptor mRNA was demonstrated by in situ hybridization using biotinylated cDNA antisense oligonucleotides. Hybridization signals were found within the seminiferous epithelium exclusively in Sertoli cell cytoplasm associated with normal spermatogenesis and in epithelia showing different signs of impairment, including SCO. It is concluded that: (1) Sertoli cells are the only cells within the seminiferous epithelium expressing FSH receptors; (2) the accumulation of FSH immunoreactivity in Sertoli cells of SCO tubules appears to be a sign of impaired Sertoli cell function.
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  • 66
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    Keywords: Carotid labyrinth ; Coexistence ; Substance P ; CGRP ; VIP ; Neuropeptide Y ; Immunohistochemistry ; Rana catesbeiana (Anura)
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    Notes: Abstract Double immunohistochemical staining with rhodamine- and fluorescein isothiocyanate (FITC)-conjugated antisera revealed the coexistence of substance P (SP) and neuropeptide Y (NPY), and SP and calcitonin gene-related peptide (CGRP) in most nerve fibers in the intervascular stroma of the carotid labyrinth of the bull-frog, Rana catesbeiana, although there were a few fibers which showed only SP- or NPY-immunoreactivity. Approximately one third of SP-immunoreactive fibers also showed coexistence with vasoactive intestinal polypeptide (VIP)-immunoreactivity, and a few fibers contained VIP without SP. The combination of the double immunofluorescence technique and alternate consecutive sections further demonstrated the possible coexistence of SP, VIP, NPY, and CGRP. This coexistence of four different peptides in the same nerve fibers was proved by the following two evident facts: 1) some SP fibers which demonstrated coexistence with NPY-immunoreactivity were assumed to be continuous with those showing VIP-immunoreactivity, and 2) almost all of the SP fibers showed coexistence with CGRP-immunoreactivity. By this reasoning, nearly one third of SP fibers may demonstrate coexistence with NPY-, VIP-, and CGRP-immunoreactivities. These multiple peptides might be involved in vascular regulatory function, which is a possible function of the amphibian carotid labyrinth.
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  • 67
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    Keywords: Key words: Pineal organ ; Neuron-specific enolase ; Immunohistochemistry ; Three-dimensional reconstruction ; Post-hatching development ; Domestic fowl
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    Notes: Abstract. Immunohistochemistry for neuron-specific enolase (NSE) revealed that NSE is localized in both a limited number of pinealocytes and intrinsic afferent neurons in the pineal organ of the domestic fowl. Furthermore, a computer-assisted three-dimensional imaging technique allowed to clarify the reverse distributional pattern of both elements: NSE-positive pinealocytes displayed a dense distribution especially in the vesicular portion of the gland, whereas NSE-immunoreactive nerve cells were mainly found in the pineal stalk. The number of NSE-positive intrinsic neurons in the pineal organ of chickens decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ. On the other hand, immunoreactive pinealocytes increased remarkably in the end-vesicle of the organ with age, followed by a gradual expansion toward the proximal portion. Thus, the spectacular increase in NSE-positive pinealocytes and the progressive reduction of reactive neurons occurred in parallel during the course of post-hatching development. NSE-immunoreactive pinealocytes displayed morphological characteristics of bipolar elements, endowed with an apical protrusion into the pineal lumen and a short basal process at younger stages, whereas multipolar types of NSE-positive pinealocytes were predominantly found in the adult domestic fowl. These results indicate that in the pineal organ of the domestic fowl (1) the ontogenetic expansion of NSE-immunoreactive pinealocytes is paralleled by a regressive afferent innervation, (2) the NSE-positive pinealocytes transform from a bipolar (columnar) type to a multipolar type during post-hatching development, and (3) these ontogenetic changes in the NSE-immunoreactivity and morphology of pinealocytes may reflect the development of a neurosecretory-like capacity of the organ.
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  • 68
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    Keywords: Key words: Pharynx ; Lung ; Calcitonin gene-related peptide ; Substance P ; Coexistence ; Immunohistochemistry ; Rana catesbeiana (Anura)
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    Notes: Abstract. Indirect double immunofluorescence labelling in the pharynx and lung of the bullfrog, Rana catesbeiana, demonstrated the occurrence, distribution, and coexistence of two neuropeptides. In the pharynx, immunoreactive calcitonin gene-related peptide (CGRP) and substance P (SP) were localized in nerve fibers distributed within and just beneath the ciliated epithelium. In the lung, CGRP and SP were localized in nerve fibers in five principal locations: 1) within the smooth muscle layer in the interfaveolar septa; 2) in the luminal thickened edges of the septa; 3) around the pulmonary vasculature; 4) within, and 5) under the ciliated epithelium. Within the smooth muscle layer in the septa, luminal thickened septa, and around blood vessels, almost all fibers showed coexistence of CGRP and SP. Within and just beneath the ciliated epithelium in the thickened septa, all fibers showed coexistence of CGRP and SP. No immunoreactivity for vasoactive intestinal polypeptide, neuropeptide Y, galanin, somatostatin, FMRFamide, and leucine- and methionine-enkephalins was detected in the nerve fibers within the larynx and the lung. Together with our previous data, the present findings suggest that peptidergic mechanisms are involved in the regulation of amphibian respiratory systems throughout their life.
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  • 69
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    Cell & tissue research 279 (1994), S. 169-181 
    ISSN: 1432-0878
    Keywords: Key words: Adrenal gland ; Dorsal root ganglia ; Immunohistochemistry ; Neurofilament ; Neuronal tracing ; Neuropeptides ; Nitric oxide synthase ; Substance P ; Guinea-pig
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    Notes: Abstract. Retrograde neuronal tracing in combination with double-labelling immunofluorescence was applied to distinguish the chemical coding of guinea-pig primary sensory neurons projecting to the adrenal medulla and cortex. Seven subpopulations of retrogradely traced neurons were identified in thoracic spinal ganglia T1-L1. Five subpopulations contained immunolabelling either for calcitonin gene-related peptide (CGRP) alone (I), or for CGRP, together with substance P (II), substance P/dynorphin (III), substance P/cholecystokinin (IV), and substance P/nitric oxide synthase (V), respectively. Two additional subpopulations of retrogradely traced neurons were distinct from these groups: neurofilament-immunoreactive neurons (VI), and cell bodies that were nonreactive to either of the antisera applied (VII). Nerve fibres in the adrenal medulla and cortex were equipped with the mediator combinations I, II, IV and VI. An additional meshwork of fibres solely labelled for nitric oxide synthase was visible in the medulla. Medullary as well as cortical fibres along endocrine tissue apparently lacked the chemical code V, while in the external cortex some fibres exhibited code III. Some intramedullary neuronal cell bodies revealed immunostaining for nitric oxide synthase, CGRP or substance P, providing an additional intrinsic adrenal innervation. Perikarya, immunolabelled for nitric oxide synthase, however, were too few to match with the large number of intramedullary nitric oxide synthase-immunoreactive fibres. A non-sensory participation is also supposed for the particularly dense intramedullary network of solely neurofilament-immunoreactive nerve fibres. The findings give evidence for a differential sensory innervation of the guinea-pig adrenal cortex and medulla. Specific sensory neuron subpopulations suggest that nervous control of adrenal functions is more complex than hitherto believed.
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  • 70
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    Cell & tissue research 279 (1994), S. 209-213 
    ISSN: 1432-0878
    Keywords: Key words: Calcium-binding proteins ; Immunohistochemistry ; Mesonephros ; Metanephros ; Chick embryo (White leghorn)
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    Notes: Abstract. The ontogeny of two calcium-binding proteins (calbindin-D28k and calretinin) was studied by immunohistochemical techniques in developing chick kidney. This study showed the presence of calbindin on the 5th incubation day and calretinin on the 7th incubation day in mesonephric distal and connecting tubules, and in the medial wall of the Wolffian duct. At later stages, immunostaining for these two proteins, in particular for calretinin, was also demonstrated in some metanephric proximal tubules. Glomeruli and Bowman's capsules were negative both in the mesonephros and metanephros. The presence of calretinin in the developing kidney has thus been demonstrated for the first time. The early expression of calbindin and calretinin in mesonephric distal tubules suggests their role in regulating the final excretion of calcium. The different patterns of immunoreactivity of the walls of the Wolffian duct can be correlated with their different histogenetic and histological features.
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  • 71
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    Cell & tissue research 276 (1994), S. 213-221 
    ISSN: 1432-0878
    Keywords: Gut-associated lymphoid tissue (GALT) ; M(membranous)-cells ; Immunohistochemistry ; Cytokeratins ; Yeast ; Pig (Minipig, Göttingen)
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    Notes: Abstract The intermediate filaments of the dome epithelium of porcine Peyer's patches were studied by immunohistochemistry. The labelling patterns of monospecific antibodies directed against cytokeratins 8, 18 and 19 differed considerably. About 40% of the dome epithelial cells were intensely labelled by three different anti-cytokeratin 18 antibodies, indicating that large amounts of cytokeratin 18 are present in these cells. In order to verify that these cytokeratin-18-immunoreactive cells were M-cells, uptake studies using fluorescein-labelled yeast particles were performed. Numerous yeast particles were found exclusively in dome epithelial cells that were highly positive for cytokeratin 18, thus representing M-cells. In contrast, the content of cytokeratin 19 in M-cells was lower than that in neighbouring enterocytes. The labelling intensity of cytokeratin 8 did not differ between M-cells and enterocytes. In addition, the absence of vimentin and desmin from the dome epithelium of porcine Peyer's patches was demonstrated. The results show (1) that porcine M-cells differ from enterocytes in the composition of their cytoskeleton, (2) that cytokeratin 18 is a useful marker for detecting porcine M-cells and (3) that this marker directly correlas with M-cell function.
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  • 72
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    Cell & tissue research 273 (1993), S. 467-474 
    ISSN: 1432-0878
    Keywords: Basic fibroblast growth factor ; Salivary glands ; Cell growth assay ; Immunohistochemistry ; Radioimmunoassay ; Rat (Wistar)
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    Notes: Abstract We studied the occurrence and localization of basic fibroblast growth factor (bFGF) in rat salivary glands using a specific monoclonal antibody. It was shown that the extract of rat salivary glands has a pronounced stimulatory activity on the growth of bovine capillary endothelial cells, which is blocked by the addition of an antibody against bFGF. The concentration of bFGF in the submandibular/sublingual gland, as determined by radioimmunoassay, was ∼80% that in the brain. Immunocytochemistry revealed bFGF-immunoreactivity localized primarily in the epithelial cells lining the striated ducts and excretory ducts of the parotid, sublingual and submandibular glands. In addition, intense bFGF-immunoreactivity was observed in the granular convoluted tubule of the submandibular gland, localized predominantly in the agranular pillar cells, which lay in small numbers among the majority of weakly immunostained cells containing many apical secretory granules. At the electron-microscopic level, the immunoreactive material was distributed diffusely in the cytoplasmic matrix and nuclei of all immunoreactive cells, whereas it was absent from all cytoplasmic organelles including the secretory granules. These results indicate that bFGF is localized in different cellular and subcellular compartments from those of other growth factors in the duct system of rat salivary glands.
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  • 73
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    Cell & tissue research 273 (1993), S. 547-555 
    ISSN: 1432-0878
    Keywords: Pancreas, endocrine ; Immunohistochemistry ; Neuropeptide Y ; Peptide YY ; FMRFamide ; LPLRFamide ; Vasoactive intestinal polypeptide (VIP) ; Oncorhynchus mykiss (Teleostei) ; Gadus morhua (Teleostei)
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    Notes: Abstract Endocrine cells exhibiting immunoreactivity to FMRFamide-like, LPLRFamide-like, neuropeptide Y(NPY)-like and peptide YY(PYY)-like peptides were found in the periphery of the Brockmann bodies of the cod, Gadus morhua, and rainbow trout, Oncorhynchus mykiss. No immunoreactivity or very weak labelling was found with antisera to pancreatic polypeptide (PP). Vasoactive intestinal polypeptide (VIP)-like immunoreactivity was found in nerve fibres, whereas labelling with VIP antiserum in endocrine cells disappeared after preincubation with nonimmune serum. There were always more immunoreactive cells in the rainbow trout than in the cod. No immunoreactivity could be seen with antisera to gastrin/cholecystokinin (CCK) or enkephalin. Double-labelling studies were performed to study the colocalization of the peptides in peripheral endocrine cells. Cells immunoreactive to NPY were also labelled with antisera to FMRFamide, LPLRFamide and PYY. The co-localization pattern of NPY varied; in some Brockmann bodies, a population of the immunoreactive cells showed co-localization and others contained NPY-like immunoreactivity only, whereas in other Brockmann bodies, all NPY-labelled cells also contained FMRFamide-like, LPLRFamide-like and PYY-like immunoreactivity. Cells immunoreactive to PYY similarly contained FMRFamide-like, LPLRFamide-like and NPY-like immunoreactivity, comparable to the patterns observed with NPY. Glucagon-like immunoreactivity was found at the periphery of the Brockmann bodies. A subpopulation of the glucagon-containing cells contained NPY-like immunoreactivity. PYY-like immunoreactivity was also found co-localized with glucagon-like immunoreactivity, as were FMRFamide-like and LPLRFamide-like immunoreactivity. Therefore, either NPY-like and PYY-like immunoreactivity together with FMRFamide-like and LPLRFamide-like immunoreactivity occur in the same endocrine cells of the Brockmann body of the cod and rainbow trout, or a hybrid NPY/PYY-like peptide recognized by both NPY and PYY antisera is present in the Brockmann body.
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  • 74
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    Cell & tissue research 274 (1993), S. 181-187 
    ISSN: 1432-0878
    Keywords: Neuropeptides ; Stellate ganglion ; Immunohistochemistry ; Pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization and distribution of neuropeptides including neuropeptide Y (NPY), [Met5]enkephalin-Arg6-Gly7-Leu8 (MEAGL), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), substance P and somatostatin (SOM) were analyzed in the stellate ganglion of the pig by use of the indirect immunofluorescence technique. NPY, MEAGL, SOM, VIP and CGRP immunoreactivities were found to exist in subpopulations of neuronal cell bodies of the stellate ganglion. A population of the small intensely fluorescent (SIF) cells showed MEAGL immunoreactivity. In addition, the presence of NPY-, MEAGL-, CGRP-, SP-, SOM- and VIP-immunoreactive nerve fibers and axonal varicosities were observed in the stellate ganglion. The localization and pattern of distribution of these peptides in the porcine stellate ganglion were compared with studies carried out on stellate ganglia of other mammalian species.
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  • 75
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    Keywords: Subcommissural organ ; Reissner's fiber ; Secretory products ; Immunohistochemistry ; Development, phylogenetic ; Class-specific epitopes ; Dogfish, Scyliorhinus canicula (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have raised antisera against extracts of the subcommissural organ (SCO) of the dogfish, Scyliorhinus canicula L. Brains of 2900 specimens were collected in acetone, and the region containing the SCO and posterior commissure was removed and extracted in three different media. Antisera against these crude extracts were raised in rats and rabbits. Sequential absorptions of the antisera with extracts from different regions of the dogfish brain were performed to eliminate unwanted antibodies. When used to immunostain sections of the whole central nervous system of the dogfish, these purified antisera reacted selectively with the SCO-Reissner's fiber complex. An antiserum against bovine Reissner's fiber was also used. The antisera against the dogfish SCO and bovine Reissner's fiber showed the same staining pattern in the SCO and the Reissner's fiber of the dogfish. For comparative purposes, the brains of 15 vertebrate species from all vertebrate classes were immunostained with both antisera. The anti-dogfish SCO serum reacted with the SCO of the dogfish and that of other phylogenetically related elasmobranch species. Neither the SCO of a primitive elasmobranch species, Heptranchias perlo, nor the SCO of the other classes of vertebrates reacted with the anti-dogfish SCO serum. However, the antiserum against bovine Reissner's fiber reacted with the SCO of all the investigated species. It is concluded that some epitopes (or compounds) in the secretory material of the SCO are class-specific, whereas others are conserved and are synthesized by the SCO in most vertebrate species.
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    Cell & tissue research 234 (1983), S. 519-531 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Serotonin ; Immunohistochemistry ; Fluorescence histochemistry ; Sympathectomy ; Dog
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Monoamine (noradrenaline and serotonin)-containing nerve fibers distributed in the pineal organ of the dog were studied by means of histochemistry (formaldehyde-induced fluorescence) and immunohistochemistry (peroxidase-antiperoxidase-PAP method) with the use of a serotonin antiserum. With the fluorescence-histochemical technique a dense network of blue-green fluorescent fibers was demonstrated in the pineal organ. Most of these fibers formed a perivascular plexus and their branches penetrated into the intercellular spaces of the parenchymal cells. Since these fibers completely disappeared ten days after bilateral removal of the superior cervical ganglia, it was confirmed that they are noradrenergic post-ganglionic sympathetic nerve fibers. A few yellow-fluorescent fibers were detected in the proximal part of the organ after ganglionectomy. By the use of the PAP method, intensively immunoreactive parenchymal cells and nerve fibers were demonstrated. The distribution pattern of these fibers was similar to that of the fluorescent sympathetic fibers. After almost all immunopositive fibers had been abolished by sympathectomy, some serotonin-containing fibers remained. The latter could be traced back to a system of serotonin fibers in the epithalamic region. These findings suggest that 1) the sympathetic noradrenergic fibers in the pineal organ of the dog take up serotonin which is released from the pinealocytes, and 2) this organ receives a dual monoamine innervation via peripheral noradrenergic and central serotonergic nerve fibers.
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    Cell & tissue research 235 (1984), S. 497-502 
    ISSN: 1432-0878
    Keywords: Prolactin cell ; Mitosis ; Sex difference ; Aging ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The anterior pituitary of colchicine-pretreated male and female rats from 20 days to 12 months of age was stained immunohistochemically with anti-rat prolactin serum. Immunoreactive mitotic cells were identified in all groups of rats. In adult female rats the mitotic index of prolactin cells was higher at oestrus than at other stages of the oestrus cycle and than that in male rats of comparable ages. If adult female rats were ovariectomized on the second day of dioestrus or on the day of proestrus, the mitotic indices at presumptive oestrus were less than those in sham-operated controls at oestrus. Estrogen administration to ovariectomized rats significantly elevated the mitotic index of prolactin cells at 48 h after the treatment. The mitotic indices of prolactin cells in female rats reached a peak at 60 days of age, and then decreased with age. In male rats the mitotic indices showed a steady decrease from the value at 20 days of age. A sex difference in the mitotic indices of prolactin cells was noted from 60 days to 12 months of age. The present results clearly demonstrate that differentiated prolactin cells can undergo mitosis and that a sex difference in the mitotic activity of prolactin cells is present during adult life.
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  • 78
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    Cell & tissue research 199 (1979), S. 25-36 
    ISSN: 1432-0878
    Keywords: Uterine proteins (rabbit) ; Uteroglobin ; Blastocyst ; Immunohistochemistry ; Gel chromatography ; Disc electrophoresis ; Immunoelectrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Proteins of uterine fluid and lung homogenates of the rabbit were separated by gel and ion exchange chromatography. Purified protein fractions were used for immunisation and antiserum production. By means of several absorptions, six monospecific antisera against uteroglobin and five other proteins were obtained. Using immunohistochemistry, four of them could be localised in the uterine epithelium from oestrus and the first and the seventh day post coitum, and also in the blastocyst. The present study indicates the involvement of different endometrial cells in the synthesis and release of the various proteins of uterine secretion.
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  • 79
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Serotonin ; Neuroendocrine regulation ; Monkey (Macaca fuscata) ; Immunohistochemistry
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    Notes: Summary The distributional pattern of serotonin-containing nerve fibers in the hypothalamus of the monkey (Macaca fuscata) was analyzed with the use of the peroxidaseantiperoxidase method in conjunction with a highly sensitive and specific anti-serotonin serum. The highest concentrations of serotonin-immunoreactive varicose fibers were found in the nucleus praeopticus medialis, nucleus ventromedialis hypothalami, and the complex of mammillary nuclei (nucleus praemamillaris, supramamillaris, mamillaris medialis et lateralis). However, the nucleus suprachiasmaticus, where numerous serotoninergic fibers have been reported to occur in the rat, appeared to be almost devoid of these fibers. The infundibular stalk, and the intermediate and posterior lobes of the pituitary contained considerable numbers of immunoreactive fibers. The present study provides a morphological basis for possible clarification of the influence of serotoninergic projections on various neuroendocrine mechanisms in primates. Furthermore, an attempt was made to clarify the differences and similarities concerning the distributional patterns of serotoninergic nerve fibers within the monkey hypothalamus in contrast to the rat hypothalamus.
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  • 80
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    Cell & tissue research 236 (1984), S. 733-735 
    ISSN: 1432-0878
    Keywords: S-100 protein ; Adrenal medulla ; Sustentacular cells ; Human fetus ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Adrenal glands of human fetuses were investigated by means of an immunohistochemical method with the use of an anti-S-100 serum. S-100-immunoreactivity was recognized in sustentacular cells located among the chromaffin cells. A characteristic circular arrangement of the immunostained cells was found in the central region of the adrenal glands. It surrounded aggregations of non-argyrophilic, small, round cells, which were identified as the remaining sympathoblasts (primitive sympathetic cells).
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    Cell & tissue research 201 (1979), S. 343-348 
    ISSN: 1432-0878
    Keywords: Merkel cell ; Sinus hair follicle ; Rat skin ; Enkephalin ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The Merkel cells from sinus hair follicles of rats were investigated by immunohistochemistry using different antisera against neuropeptides and gastroenteropancreatic (GEP)-hormones. For the first time it has been demonstrated that Merkel cells exhibit an immunoreactivity towards metenkephalin (methionine-enkephalin). The met-enkephalin immunoreactivity was restricted to Merkel cells and was not found in associated nerve axons or terminals. Denervation of Merkel cells did not affect the met-enkephalin immunoreactivity. Antisera against leu-enkephalin (leucine-enkephalin) and other polypeptides did not produce an immunoreaction. The demonstration of met-enkephalin-like immunoreactivity supports the concept that the Merkel cell is a member of the paraneuronal system and a potential neuroreceptor cell.
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  • 82
    ISSN: 1432-0878
    Keywords: Immunohistochemistry ; Paraganglia ; Aging ; Catecholamines ; Catecholamine-synthesizing enzymes
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    Notes: Summary The catecholamine-synthesizing enzymes, tyrosine hydroxylase, dopamine-β-hydroxylase and phenylethanolamine-N-methyltransferase were examined by immunohistochemistry in hypertrophied paraganglia of aged male Fischer-344 rats. All paraganglionic cells reacted with antibodies against tyrosine hydroxylase. Dopamine β-hydroxylase was identified in most paraganglionic cells, indicating that they synthesized norepinephrine. A variable number of paraganglia were positive for phenylethanolamine-N-methyltransferase, which suggested that they synthesized epinephrine. The formaldehyde-induced fluorescence method demonstrated greenish-yellow fluorescence or yellowish-brown fluorescence. The intensity of the fluorescence was in the same range as in adrenal medullary cells. The observations indicate that paraganglia are capable of synthesizing epinephrine.
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  • 83
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    Keywords: Serotonin ; Neuropeptides ; Endocrine cells ; Autonomic innervation ; Gut ; Rectal gland ; Immunohistochemistry ; Chimaera monstrosa (Holocephala)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The occurrence and distribution of endocrine cells and nerves were immunohistochemically demonstrated in the gut and rectal gland of the ratfish Chimaera monstrosa (Holocephala). The epithelium of the gut mucosa revealed open-type endocrine cells exhibiting immunoreactivity for serotonin (5HT), gastrin/cholecystokinin (CCK), pancreatic polypeptide (PP)/FMRFamide, somatostatin, glucagon, substance P or gastrin-releasing peptide (GRP). The rectum contained a large number of closed-type endocrine cells in the basal layer of its stratified epithelium; the majority contained 5HT- and GRP-like immunoreactivity in the same cytoplasm, whereas others were immunoreactive for substance P. The rectal gland revealed closed-type endocrine cells located in the collecting duct epithelium. Most of these contained substance P-like immunoreactivity, although some reacted either to antibody against somatostatin or against 5HT. Four types of nerves were identified in the gut and the rectal gland. The nerve cells and fibers that were immunoreactive for vasoactive intestinal peptide (VIP) and GRP formed dense plexuses in the lamina propria, submucosa and muscular layer of the gut and rectal gland. A sparse network of gastrin- and 5HT-immunoreactive nerve fibers was found in the mucosa and the muscular layer of the gut. The present study demonstrated for the first time the occurrence of the closed-type endocrine cells in the mucosa of the rectum and rectal gland of the ratfish. These abundant cells presumably secrete 5HT and/or peptides in response to mechanical stimuli in the gut and the rectal gland. The peptide-containing nerves may be involved in the regulation of secretion by the rectal gland.
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  • 84
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    Cell & tissue research 260 (1990), S. 307-314 
    ISSN: 1432-0878
    Keywords: Astroglial cells ; Ependymocytes ; Spinal cord ; Immunohistochemistry ; Bufo bufo L. (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunohistochemical and ultrastructural techniques have been used to demonstrate glial fibrillary acidic protein (GFAP) immuno-positive cells in the adult toad spinal cord. Two types of GFAP-immunoreactive cells were observed: ependymocytes and radial astrocytes. GFAP-positive ependymocytes were scarce and contained the immunoreactive product in their processes. They showed intermediate filaments in the basal pole and in their processes when studied with the electron microscope. These immuno-positive ependymocytes represent the tanycytic form of ependymal cells because their processes ended at the subpial zone. The radial astrocytes showed a more intensive immunoreactive product in somata and processes when they were located far away from the ependymal layer. Cell bodies and processes were also associated with blood vessels, but most of the processes ended at the subpial zone forming a continuous subpial glia limitans. The GFAP-positive processes, which form this subpial glia limitans in the toad spinal cord, belong to both tanycytic ependymocytes and radial astrocytes, whose somata are located in the grey matter. These findings lead us to suggest that both types of GFAP-immunopositive cells might be the functional equivalents of mammalian astrocytes.
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  • 85
    ISSN: 1432-0878
    Keywords: PGP 9.5 (human protein gene product 9.5) ; Skin ; Immunohistochemistry ; Nerve fibres ; Dendritic cells ; Merkel cells ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Sections of human skin were processed according to the indirect immunofluorescence technique with a rabbit antiserum against human protein gene product 9.5 (PGP 9.5). Immunoreactivity was detected in intraepidermal and dermal nerve fibres and cells. The intraepidermal nerves were varicose or smooth with different diameters, running as single processes or branched, straight or bent, projecting in various directions and terminating in the stratum basale, spinosum or granulosum. The density of the intraepidermal nerves varied between the different skin areas investigated. PGP 9.5-containing axons of the lower dermis were found in large bundles. They separated into smaller axon bundles within the upper dermis, entering this portion of the skin perpendicular to the surface. Then they branched into fibres mainly arranged parallel to the epidermal-dermal junctional zone. However, the fibres en route to the epidermis traversed the upper dermis more or less perpendicularly. Furthermore, immunoreactive dermal nerve fibres were found in the Meissner corpuscles, the arrector pili muscles, hair follicles, around the eccrine and apocrine sweat glands and around certain blood vessels. Such fibres were also observed around most subcutaneous blood vessels, sometimes heavily innervating these structures. Numerous weakly-to-strongly PGP 9.5-immunoreactive cells were found both in the epidermis and in the dermis.
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  • 86
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    Cell & tissue research 261 (1990), S. 85-96 
    ISSN: 1432-0878
    Keywords: Sensilla ; Scolopidia ; Thermoreceptors ; Hygroreceptors ; Actin filaments ; Cryofixation ; Highpressure freezing ; Immunohistochemistry ; Antheraea pernyi, Periplaneta americana, Schedorhinotermes lamanianus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Two types of insect sensilla, mechanosensitive scolopidia and thermo-/hygrosensitive poreless sensilla contain a scolopale, which consists of numerous microtubules embedded in bundles of filaments (7–10 nm in diameter). The bundles are readily seen in the electron microscope in cryofixed (high-pressure freezing and rapid injection) and substituted samples. The filaments can be identified as actin filaments by using fluorescent phalloidins. Both electron microscopy and Triton-extraction exeriments reveal mechanical linkage between the main components in both types of sensilla. Since myosin appears to be absent in the scolopale, the actin filaments are unlikely to be involved in any contraction mechanism; these filaments more probably provide mechanical stability. The functional properties of the scolopale are discussed.
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  • 87
    ISSN: 1432-0878
    Keywords: Atrial natriuretic peptide (ANP) ; Chicken natriuretic peptide (chANP) ; Atrial-specific granule ; Sarcoplasmic reticulum ; Immunohistochemistry ; Domestic fowl (Aves, Phasianiformes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An immunohistochemical study using antiserum raised against synthetic chicken natriuretic polypeptide was used to investigate the distribution of this peptide in the chicken heart. Immunoreactive cells, both in the atrial and ventricular walls, were identified by electron microscopy, and electron-dense granules in the atrial and ventricular cardiocytes were revealed to be storage sites of the peptide. The electron-dense material, thought to be the peptide, was found in the sarcoplasmic reticulum, and it is suggested that a secretory pathway of the peptide through the latter to extracellular space, may be present, in addition to an exocytotic one.
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  • 88
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    Cell & tissue research 259 (1990), S. 313-319 
    ISSN: 1432-0878
    Keywords: Opioid peptides ; Serotonin ; Enterochromaffin cells ; Endocrine pancreas ; Immunohistochemistry ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous immunochemical investigations have demonstrated various opioid peptides in the pancreas. However, controversies exist related to the cellular localization of these peptides in the endocrine pancreas. Therefore, the guinea pig endocrine pancreas was immunohistochemically investigated for the presence of opioid peptides derived from pro-dynorphin, pro-enkephalin or pro-opiomelanocortin. Immunoreactivities were demonstrated on serial semithin sections by the peroxidase anti-peroxidase technique. In routinely immunostained sections, immunoreactivities for dynorphin A and α-neo-endorphin were localized in pancreatic enterochromaffin cells, but not in islet cells. Immunoreactivity for Met-enkephalin was confined exclusively to B-cells and was localized only in some secretory granules. However, pre-treatment of semi-thin sections with trypsin and carboxypeptidase B led to a marked increase of Met-enkephalin immunoreactivity in B-cells. In addition, immunoreactivities for Met-enkephalin-Arg-Gly-Leu and bovine adrenal medulla dodecapeptide could be demonstrated in B-and A-cells, and β-endorphin immunoreactivity was localized in A-cells. In no case, however, were immunoreactivities detected for bovine adrenal medulla docosapeptide, peptide F, corticotropin, melanotropin or dynorphin 1–32. The immunohistochemical findings indicate that opioids of different peptide families are present in the guinea pig endocrine pancreas. Since several opioid peptides of the corresponding pro-hormones could be demonstrated in the reference organs but not in the pancreas, it is concluded that the biosynthetic pathways of the respective precursors are different from those in the adrenal medulla or in the pituitary.
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  • 89
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    Cell & tissue research 261 (1990), S. 333-338 
    ISSN: 1432-0878
    Keywords: Cell proliferation ; Anterior pituitary ; Postnatal development ; Immunohistochemistry ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cellular proliferation in the anterior pituitary of 2-, 8-, 15- and 30-day-old rats was examined by injection of bromodeoxyuridine 1 h before autopsy. Bromodeoxyuridine incorporated into DNA was detected immunohistochemically by use of a monoclonal antibody. The highest rate of cell proliferation was found in 2-day-old animals; it decreased thereafter during the postnatal period. Possible toxic effects of colchicine on cellular proliferation were examined. Colchicine treatment (10 mg/kg in 8- and 30-day-old animals) significantly decreased the number of bromodeoxyuridine-labelled cells/mm2 in 8-day-old rats. Some sections were doubly immunostained for bromodeoxyuridine and various pituitary hormones. The proportion of doubly-immunostained cells to all proliferating cells was generally low, ranging from 23% at 2 days to 32% at 30 days of age.
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  • 90
    ISSN: 1432-0878
    Keywords: Parathyroid glands ; Neuropeptide colocalization ; Calcitonin gene-related peptide (CGRP) ; Substance P ; Immunohistochemistry ; Cow
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Although peptide neurotransmitters have been shown to modulate hormone secretion in many glands, there are very few studies of neurotransmitters in the parathyroid gland. Bovine parathyroid glands were collected at a local abattoir, fixed with paraformaldehyde, sectioned using a cryostat, and stained by indirect immunohistochemistry for calcitonin gene-related peptide and substance P. We were able to positively identify both neuropeptides. Nerve fibres containing calcitonin gene-related peptide and substance P were identified in contact with the tunica media of arteries and arterioles and dispersed throughout the stroma of the gland. While many of the fibres encircled parenchymal lobules, no intimate contact with the peripheral chief cells was observed. All immunoreactive fibres were found to contain both neuropeptides. Since calcitonin gene-related peptide and substance P are vasodilators, they may increase blood flow within the gland. In addition, the neuropeptides may diffuse from perilobular nerve fibres into the parenchyma, thereby modulating secretion of parathyroid hormone.
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  • 91
    ISSN: 1432-0878
    Keywords: Calbindin ; Neurohypophysis ; Development, ontogenetic ; Immunohistochemistry ; In-situ hybridization ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Spot 35 protein is a Ca-binding protein originating from the rat cerebellum; it is now referred to spot 35-calbindin. This protein is expressed in immature pituicytes of the neurohypophyseal anlage in the E11–E18 rat embryo. The gene expression of spot 35-calbindin was detected by in-situ hybridization analysis only at stage E11–E12. Profiles of spot 35-positive nerve fibers of a neurosecretory nature were found in anlage at stage E16. At this stage, some immature pituicytes are partially immunopositive for spot 35-calbindin only in their peripheral cytoplasm; others are immunonegative. At birth and thereafter through adulthood, abundant nerve fibers are the sole structures immunoreactive for spot 35-calbindin; all the pituicytes are immunonegative, resulting in a light-microscopic appearance of numerous immunonegative round profiles, corresponding to pituicytes, and capillaries embedded in the granularly immunostained neurohypophysis. The present findings suggest that, during specific embryonic stages, immature pituicytes exert some as yet unidentified roles related to Ca-mediated functions involving the expression of spot 35-calbindin.
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  • 92
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    Cell & tissue research 266 (1991), S. 391-398 
    ISSN: 1432-0878
    Keywords: Photoreceptor cells ; Nervous system central ; Visual system ; Histamine ; Transmitter ; Immunohistochemistry ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In this study, immunohistochemistry on cryostat sections is used to demonstrate anti-histamine immunoreactivity in the Drosophila brain. The results support earlier findings that histamine is probably a transmitter of insect photoreceptors. It is further shown that, in Drosophila, all imaginal photoreceptors including receptor type R7 are anti-histamine immunoreactive, whereas the larval photoreceptors do not seem to contain histamine. In addition to the photoreceptors, fibres in the antennal nerve and approximately 12 neurons in each brain hemisphere show strong histamine-like immunoreactivity. These cells arborize extensively in large parts of the central brain.
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  • 93
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    Cell & tissue research 259 (1990), S. 43-49 
    ISSN: 1432-0878
    Keywords: Lectin ; Smooth muscle cells ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunohistochemical localization of 14 kDa β-galactoside-binding lectin in various organs of adult rat was achieved using a monospecific antibody raised against lectin purified from rat lung. The antibody-stained cells were formed into small aggregates, thin fascicles, or thick bundles in the walls of blood vessels, gastrointestinal tracts and urogenital organs. From the patterns of distribution, as well as their organization, these immunoreactive cells were regarded as smooth muscle cells. This was confirmed by a double immunofluorescence study using a mixture of anti 14 kDa lectin and anti α-smooth muscle-specific actin antibodies. Strong 14 kDa lectin immunoreactivity was seen in the pericellular matrix of smooth muscle cells in intact organs as well as in detergent-treated organs from which all cellular components were extracted. From these findings, it is suggested that the 14 kDa lectin may be externalized by smooth muscle cells into their pericellular matrix and participate in the crosslinking of the complementary glycoconjugate(s) localized at that site. The macromolecular complex of glycoconjugates thus formed around smooth muscle cells may play a role in anchoring smooth muscle cells to the pericellular connective tissue thereby permitting the force of muscle contraction to be efficiently transmitted to the surrounding connective tissue proper.
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  • 94
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    Cell & tissue research 259 (1990), S. 305-312 
    ISSN: 1432-0878
    Keywords: Type-VI collagen ; Type-IV collagen ; Laminin ; Fibronectin ; Long-spacing collagen ; Immunohistochemistry ; Uvea ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of type-VI collagen in the human iris and ciliary body was investigated by means of immunohistochemical techniques and compared with that of type-IV collagen, fibronectin and laminin. As has been described for other tissues, type-VI collagen surrounds type-I and-III collagen fibers. The aggregated from of type-IV collagen (the “long-spacing” or “curly” collagen), which has already been described in the trabecular meshwork and sclera, was also observed at the ciliary muscle tips surrounding the anterior elastic tendons of this muscle. In addition, staining for type-VI collagen was seen directly adjacent to the basement membranes of the ciliary muscle cells, the iris muscles, the uveal vascular endothelia and nerves, but not adjacent to the epithelial basement membranes. The staining did not form a discrete line like the immunoreaction for type-IV collagen, but bundles of marked fibrils extended into the surrounding connective tissue. We assume that type-VI collagen similar to type-VII collagen forms part of an anchoring system for these tissues. As type-VII collagen has been described only in connection with epithelial basement membranes, both type-VI and type-VII collagens may represent anchoring fibrils, however for different tissue components.
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  • 95
    ISSN: 1432-0878
    Keywords: Mucosa-associated lymphoid tissue ; Nasal cavity ; Immunohistochemistry ; Enzyme histochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Lymphocyte and macrophage subpopulations and the stroma of mucosa-associated lymphoid tissue in the nasal cavity of the rat were examined by application of immunohistochemical and enzyme histochemical methods to cryostat sections. Nasal-associated lymphoid tissue was composed of a loose reticular network with lymphocytes and macrophages, covered by epithelium. The epithelium was infiltrated with B cells, T helper (W3/13-positive) and T suppressor/cytotoxic or large granular cells (OX8-positive), ED1-positive macrophages and Ia-positive cells. The B cell areas were populated by B cells, immunopositive for surface IgM or IgG. B cells with surface IgA or IgE were rare. Germinal centres were found infrequently. T helper cells were scattered throughout the B cell area. A few ED1-positive macrophages and ED5-positive follicular dendritic cells were observed. Strong Ia staining (mostly of B cells) was found in this area. The T cell areas contained T helper and T suppressor/cytotoxic cells in about equal amounts, and numerous ED1-positive macrophages. ED1 staining was also found in the subepithelial area. Numerous ED1-, ED2- and ED3-positive macrophages were found in the border between the lymphoid mass and the surrounding connective tissue. A few non-lymphoid cells showed weak acid phosphatase or non-specific esterase activity. The morphological observations suggest that nasal-associated lymphoid tissue plays an important role in the first contact with inhaled antigens.
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  • 96
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Differentiation ; Transplantation ; Immunohistochemistry ; GFAP ; S100 protein ; Serotonin ; Rat (Sprague-Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The rat subcommissural organ (SCO), principally composed of modified ependymocytes (a type of glial cell), is a suitable model for the in vivo study of glial differentiation. An immunohistochemical study of the ontogenesis of rat SCO-ependymocytes from embryonic day 13 to postnatal day 10 shows that these cells express transitory glial fibrillary acidic protein (GFAP) from embryonic day 19 until postnatal day 3. However, S100 protein (S100) is never expressed in the SCO-cells, contrasting with the ventricle-lining cells of the third ventricle, which contain S100 as early as embryonic day 17. Environmental factors could be responsible for the repression of GFAP and S100 in adult rats, because GFAP and S100 are observed in ependymocytes of SCO 3 months after being grafted from newborn rat into the fourth ventricle of an adult rat. Neuronal factors might be involved in the control of the expression of S100, since after the destruction of serotonin innervation by neurotoxin at birth, S100 can be observed in some SCO-ependymocytes of adult rats. On the other hand, GFAP expression is apparently not affected by serotomin denervation, suggesting the existence of several factors involved in the differentiation of SCO-cells.
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  • 97
    ISSN: 1432-0878
    Keywords: Atrial natriuretic peptide ; Brain natriuretic peptide ; C-type natriuretic peptide ; Heart ; Brain, vertebrate ; Immunohistochemistry ; Squalus acanthias (Elasmobranchii) ; Myxine glutinosa (Cyclostomata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The avidin-biotin peroxidase technique was used to determine the distribution of natriuretic peptides in the hearts and brains of the dogfishSqualus acanthias and the Atlantic hagfishMyxine glutinosa. Three antisera were used: one raised against porcine brain natriuretic peptide which cross-reacts with atrial natriuretic and C-type natriuretic peptides (termed natriuretic peptide-like immunoreactivity); the second raised against porcine brain natriuretic peptide which cross-reacts with C-type natriuretic peptide, but not with atrial natriuretic peptide (termed porcine brain natriuretic peptide-like immunoreactivity); and the third raised against rat atrial natriuretic peptide (termed rat atrial natriuretic peptide-like immunoreactivity). Only natriuretic peptide-like immunoreactivity was observed in the heart ofS. acanthias which was most likely due to the antiserum cross-reacting with C-type natriuretic peptide. No immunoreactivity was found in theM. glutinosa heart. In the brain ofS. acanthias, natriuretic peptide-like immunoreactive fibres were located in many areas of the telencephalon, diencephalon, mesencephalon, rhombencephalon, and spinal cord. Extensive immunoreactivity was observed in the hypothalamo-hypophyseal tract and the neurointermediate lobe of the hypophysis. Natriuretic peptide-like immunoreactive perikarya were found in ventromedial regions of the telencephalon and in the nucleus preopticus. Most perikarya had short, thick processes which extended toward the ventricle. Another group of perikarya was observed in the rhombencephalon. Porcine brain natriuretic peptide-like immunoreactive fibres were observed in the telencephalon, diencephalon, mesencephalon, and rhombencephalon, but perikarya were only present in the preoptic area. In theM. glutinosa brain, natriuretic peptide-like immunoreactive fibres were present in all regions. Immunoreactive perikarya were observed in the pallium, primordium hippocampi, pars ventralis thalami, pars dorsalis thalami, nucleus diffusus hypothalami, nucleus profundus, nucleus tuberculi posterioris, and nucleus ventralis tegmenti. Procine brain natriuretic peptide-like immunoreactive perikarya and fibres had a similar, but less abundant distribution than natriuretic peptide-like immunoreactive structures. Although the chemical structures of natriuretic peptides in the brains of dogfish and hagfish are unknown, these observations show that a component of the natriuretic peptide complement is similar to porcine brain natriuretic peptide or porcine C-type natriuretic peptide. The presence of natriuretic peptides in the brain suggest they could be important neuromodulators and/or neurotransmitters. Furthermore, there appears to be divergence in the structural forms of natriuretic peptides in the hearts and brains of dogfish and hagfish.
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  • 98
    ISSN: 1432-0878
    Keywords: Basic fibroblast growth factor ; Regeneration ; Degeneration ; Immunohistochemistry ; Electron microscopy ; Masseter muscle ; Myoneural junction ; Mouse (dystrophic mdx)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of basic fibroblast growth factor (bFGF)-like immunoreactivity in the masseter muscle of dystrophic mdx mice on postnatal day 28 was investigated by immunoblot analysis and electron microscopy. Crude homogenate of the masseter muscle, when subjected to immunoblotting with a bFGF antiserum, exhibited a main band with the same molecular weight (18 kDa) as bovine bFGF. By electron microscopy, bFGF immunoreactivity was detected in small regenerating myocytes; the smaller cells were the premature myocytes, the most intense staining was the immunoreactivity within the cytoplasm. Putative precursors of the muscle cells with a few myofilaments, which were most intensely labeled with anti-bFGF, contacted each other and possibly developed into multinucleated myocytes through cell fusion. Mature myocytes with densely packed myofilaments and peripherally located nuclei did not exhibit bFGF immunoreactivity; they formed myoneural junctions with motor nerve endings immunoreactive for bFGF. Early differentiating myocytes with intense bFGF-like immunoreactivity did not make contact with immunoreactive nerve terminals. Degenerating large myocytes with a limited number of distorted and/or disrupted myofilaments exhibited electron-dense deposits in the cristae of mitochondria; these deposits were not abolished by immunoadsorption control experiments. Thus, the cell-size-dependent decrease in bFGF immunoreactivity in regenerating but not in degenerating myocytes provides a morphological basis for an autoregulatory role of bFGF in muscle regeneration. This study suggests that neuronal bFGF is not involved in initial muscle regeneration in the dystrophic mdx mouse.
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  • 99
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    Cell & tissue research 267 (1992), S. 233-239 
    ISSN: 1432-0878
    Keywords: Development, ontogenetic ; Kidney ; Glomerulus ; Distal convoluted tubule (DCT) ; Histamine ; Immunohistochemistry ; Hybridization, in situ ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An antiserum against conjugated histamine and two oligonucleotide probes that detect the mRNA encoding L-histidine decarboxylase (HDC) involved in histamine synthesis were used to study the appearance of histamine and its location in the kidneys of fetal, newborn and young postnatal rats and in the kidneys of pregnant rats. On embryonic days 16 and 18 (E16 and E18), some HA-immunoreactive (HA-ir) cells were found within the largest S-shaped bodies. Histamine was found to appear rapidly between the 18th and 20th embryonic days in the convoluted tubules of the kidneys. On postnatal day 0 (P0), the distal convoluted tubules and collecting ducts exhibited bright fluorescence, the intensity of which decreased quickly so that it was faint on day P4 and absent at later stages. In kidneys of pregnant rats HA-ir was found in the epithelium of both the Bowman's capsule, collecting ducts and in a few cells within the tubules. Nonuniform HA-ir was also detected within glomeruli. No evidence for the presence of L-histidine decarboxylase mRNA in kidneys of fetuses or pregnant rats was seen. It is concluded that distinct structures in the developing rat kidney contain histamine during a period around birth from day E20 to day P4. In the pregnant rat, the epithelium that is in direct contact with the urine flow is immunoreactive for histamine from day 16 to 20 of pregnancy. The results suggest that histamine is not synthesized locally in the kidneys but rather originates from other tissues.
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  • 100
    ISSN: 1432-0878
    Keywords: Vasoactive intestinal polypeptide ; Neuropeptide Y ; Enkephalin ; Calcitonin gene-related peptide ; Galanin ; Immunohistochemistry ; Enteric nervous system ; Oreochromis mossambicus, Carassius auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of bioactive peptides in the gut and their possible electrophysiological effects on the intestinal epithelium were studied in two teleost species, the tilapia (Oreochromis mossambicus) and the goldfish (Carassius auratus). Vasoactive intestinal polypeptide-like immunoreactive nerve fibres were found beneath the intestinal epithelium of both species. Galanin-, metenkephalin-and calcitonin gene-related peptide-like immunoreactive nerve fibres were found exclusively in the mucosa of the tilapia. Both species had vasoactive intestinal polypeptide-, enkephalin- or neuropeptide Y-like immunoreactive endocrine cells; calcitonin gene-related peptide-like immunoreactive endocrine cells were additionally found in the tilapia. Somatostatin- and dopamine-β-hydroxylase-like immunoreactivities were not observed. Nerve cell bodies in the myenteric plexus of both species showed immunoreactivity for calcitonin gene-related peptide-, vasoactive intestinal polypeptide-, and galanin-like peptide. Enkephalin-like immunoreactive nerve cell bodies were present in the tilapia only. None of the peptides had a pronounced electrogenic effect. However, calcitonin gene-related peptide added to stripped intestinal epithelium of the tilapia, reduced the ion selectivity, and addition of galanin increased the ion selectivity. In goldfish intestine, both galanin and calcitonin gene-related peptide were without effect. Enkephalin counteracted the serotonin-induced reduction of the ion selectivity of the goldfish intestinal epithelium, but had no effect on the tilapia epithelium. In both species, vasoactive intestinal polypeptide reduced the ion selectivity of the intestinal epithelium, and neuropeptide Y induced an increase of the ion selectivity. Somatostatin showed no effect on the epithelial ion selectivity of either species. Tetrodotoxin did not inhibit the effects of the peptides studied. The changes in ion selectivity suggest that the enterocytes may be under the regulatory control of these peptides.
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