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  • 101
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 248 (1987), S. 297-303 
    ISSN: 1432-0878
    Keywords: Uterus (myometrium) ; Gap junctions ; Pregnancy ; Morphometry ; Freeze-fracturing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In uterine smooth muscles, gap junction plaques rapidly form during the final stages of gestation. To investigate the related mechanisms, regional differences in myometrial gap junction development in rat uterus were examined quantitatively during delivery, using thin-section and freeze-fracture techniques in combination with light- and electron microscopy. Examination of implanted and nonimplanted horns in the unilaterally ligated rat bicornuate uteri, revealed no differences in the occurrence of gap junction plaques, but after 2 to 4 pups had been delivered, the contracted segments contained more gap junction plaques than did noncontracted segments examined immediately before delivery. In all segments, gap junctions were found more frequently in the circular muscle layers than in the longitudinal muscle layers. Gap junctions ranged in size from 0.002 μm2 to 0.52 μm2, but two-thirds were less than 0.1 μm2. The frequency of small gap junction plaques (less than 0.1 μm2) was higher in the noncontracted segment. These results suggest that gap junctions are dynamic structures, and that their formation is controlled not only by general hormonal factors, possibly involved in gap junction increases in the myometrium before delivery, but also by local factors, possibly related to the contraction, that may accelerate an increase in gap junction formation during delivery.
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  • 102
    ISSN: 1432-0878
    Keywords: Epididymis ; Basement membrane ; Anionic sites ; Differentiation ; Androgen ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Anionic binding sites in the lamina densa of the basement membrane of the rat epididymal epithelium were demonstrated ultrastructurally with the use of cationized polyethyleneimine (PEI). Enzyme digestion with heparitinase removed the anionic sites, indicating that they consist largely of heparan sulfates. The anionic sites are present as early as the 16th day of gestation on the interstitial face of the lamina densa; later during gestation they are localized on both faces of the lamina densa without further modification after birth. The distribution of the anionic sites was identical all along the epididymal duct. After castration and ligation of efferent ducts or in the state of cryptorchidism the sites were more numerous and located inside the thicker portion of the lamina densa. These alterations were more prominent in the initial segment compared to the distal segments, suggesting a differential androgen dependence of the reactive sites and their patterns of distribution.
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  • 103
    ISSN: 1432-0878
    Keywords: Osteogenesis ; Marrow stromal precursors ; Collagen ; Culture in vivo ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Diffusion chambers containing bone marrow cells from adult rats were implanted intraperitoneally into rat hosts and cultured in vivo for up to 64 days. Biochemical and histological analyses of the contents of the chambers demonstrate that a connective tissue consisting of bone, cartilage and fibrous tissues is formed by precursor cells present in marrow stroma. The amounts of osteogenic tissue and DNA are directly correlated with time of implantation and with number of cells inoculated. In the chambers there is initial formation of fibrous tissue which is strongly reactive to collagen type III, laminin and fibronectin. In areas of osteogenesis which appear later within this fibrous anlage, expression of collagen type III, laminin and fibronectin decrease and collagen types I and II increase in association with bone and cartilage respectively. Where osteogenesis does not develop, fibrous tissue continues to express collagen type III. The sequential expression of the different extracellular matrix components is similar to that previously observed during osteogenic differentiation in embryonic and adult developmental systems. It is concluded that the formation of fibrous and osteogenic tissues in diffusion chambers by precursor cells present in adult marrow, resembles the normal developmental process.
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  • 104
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 250 (1987), S. 197-204 
    ISSN: 1432-0878
    Keywords: Third ventricle ; Intraventricular vessels ; Recess ; Development ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light and electron microscopy of the hypothalamic ventricle in neonatal rats demonstrate morphological specializations of the ventricular wall at the level of the premammillary region of the third ventricle. The morphological features are: (1) A ventricular recess that we have called the “hypothalamic dorso-lateral recess” (HDR). (2) The presence of intraventricular capillaries near the dorsolateral recess. (3) The HDR possessing a specialized ependymal lining; this consists of non-ciliated cells with short microvilli and bleb-like processes. (4) The existence of cerebrospinal fluid-contacting neurons within the HDR. (5) The presence of numerous phagocytic supraependymal cells. The HDR is not found in adult rats. This indicates that the dorso-lateral recess may play a physiological role during development.
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  • 105
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 250 (1987), S. 323-330 
    ISSN: 1432-0878
    Keywords: Tunicamycin ; β-Receptor ; Parotid gland ; Isoproterenol ; Secretion ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological and functional effects of tunicamycin were studied in rat parotid glands at the stage of the reformation of secretory granules following secretory stimulation by isoproterenol. Tunicamycin inhibited the incorporation of (3H)-mannose into the acid-insoluble fraction but had no effect on total protein synthesis as determined by the incorporation of (14C)-leucine. Thus the administration of tunicamycin in vivo inhibits the synthesis of mannose-rich glycoproteins in a manner similar to that in an in vitro system. The ultrastructure of the acinar cell showed little change following treatment with this drug, except that the number of reaccumulated secretory granules was greater than in the control. Amylase secretion stimulated by isoproterenol was inhibited in tunicamycin-treated cells, but did not decrease following treatment with N6,2′-O-dibutyryladenosine 3′-5′-cyclic monophosphate, a secretory stimulator bypassing the β-receptor. A radio-receptor assay using (3H)-dihydroalprenolol and direct localization using the fluorescent β-adrenergic blocker 9-amino-acridin propranolol showed a marked reduction in the binding activity of β-receptor following treatment with tunicamycin. Thus the inhibition of N-linked glycosylation appears to produce profound effects on the β-adrenergic receptor-adenylate cyclase complex of acinar cells, although the steps of the transport and the exocytotic discharge of secretory materials are not affected.
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  • 106
    ISSN: 1432-0878
    Keywords: Neurofilaments ; Glial fibrillary acidic protein ; Sympathetic ganglia ; Intraocular transplantation ; Rat ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of neurofilament (NF)-like and glial fibrillary acidic protein (GFAP)-like immunoreactivities was studied in sympathetic ganglia of adult rats and guinea pigs during normal conditions and after perturbation. In the superior cervical ganglion (SCG) of normal rats, many ganglion cells and nerve fibers show NF immunoreactivity. Some of these nerve fibers disappear after preganglionic decentralization of SCG; this indicates the presence of a mixture of preand postganglionic NF-positive nerves in the ganglion. Cuts in both preand postganglionic nerves result in a marked increase in GFAP immunoreactivity in SCG, whereas NF immunoreactivity increases in nerve cell bodies after preganglionic cuts. Only a few ganglion cells show NF immunoreactivity in the normal SCG of guinea pig. All intraganglionic NF-positive nerves are of preganglionic origin; decentralization abolishes NF immunoreactivity in these nerve fibers. The inferior mesenteric ganglion, the hypogastric nerves and colonic nerves in guinea pigs contain large numbers of strongly NF-immunoreactive nerve fibers. When the SCG of adult rat is grafted to the anterior eye chamber of adult rat recipients, both ganglionic cell bodies and nerve fibers, forming on the host iris from the grafted ganglion, are NF-positive. As only the perikarya of these neurons normally exhibit NF immunoreactivity, and the terminal iris arborizations are NF-negative, it appears that the grafting procedure causes NF immunoreactivity to become more widespread in growing SCG neurons.
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  • 107
    ISSN: 1432-0878
    Keywords: Serotonin ; Neuropeptide Y ; Suprachiasmatic nucleus ; Radioautography ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The morphological substrate of putative serotonin (5-HT)/neuropeptide Y (NPY) interactions in thé suprachiasmatic nucleus (SCN) was investigated by combined radioautography and immunocytochemistry after intraventricular administration of (3H)5-HT in the rat. In the ventral portion of the SCN, the distribution of (3H)5-HT uptake sites overlapped closely the NPY-immunoreactive terminals. Previous investigations have shown that the dense 5-HT and NPY innervations of the SCN originate in different structures, i.e., the midbrain raphe nuclei and the ventral lateral geniculate nucleus, respectively. Accordingly, in the present study, destruction of 5-HT afferents by 5,7-dihydroxytryptamine was not found to induce any modification in NPY staining and, in ultrastructural immuno-radioautographic preparations, two distinct pools of axonal varicosities could be identified. Both 5-HT and NPY terminals established morphologically defined synaptic junctions, sometimes on the same neuronal target. Some cases of direct axo-axonic appositions between the two types of terminals were also encountered. These data constitute additional criteria for characterizing the cytological basis of the multiple transmitter interactions presumably involved in the function of the SCN as a central regulator of circadian biological rhythms.
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  • 108
    ISSN: 1432-0878
    Keywords: Anterior pituitary gland ; FSH cells ; LH cells ; Development ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Serum concentration of follicle-stimulating hormone (FSH) in the juvenile female rat increases independently from that of luteinizing hormone (LH). The objective of this study was to determine whether this increase in serum FSH is accompanied by a proliferation of FSH-cells greater than the proliferation of LH-cells. Thus, we measured circulating FSH and LH in female rats on days 3, 10, 13, 17, and 20, calculated the percentages of adenohypophyseal cells that contained FSH or LH on days 3, 10, and 20, and determined whether cells containing only FSH existed on day 10. Serum FSH concentrations on days 10 and 13 were significantly greater than those on days 3, 17, or 20. No differences existed in serum LH concentrations. Cells containing FSH or LH were distributed throughout the entire adenohypophyses of 3, 10, and 20-day-old females. Clusters of these cells were observed in the ventral regions of adenohypophyses of 3-day-old females. The percentages of adenohypophyseal cells containing FSH increased significantly from ∼9% in 3-day-old rats to ∼17% in 10-day-old rats and then decreased to ∼14% in 20-day-old animals. At all ages the percentages of adenohypophyseal cells containing FSH were similar to the percentages of cells containing LH. At 10 days of age, all cells containing FSH also contained LH and all cells containing LH also contained FSH. These data suggest that the increase in serum FSH in the juvenile female rat is associated with an increase in the percentage of adenohypophyseal cells containing FSH and that at this time all cells containing FSH also contain LH.
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  • 109
    ISSN: 1432-0878
    Keywords: Primary immune response ; Thymus-independent antigens ; Development, ontogenetic ; Lymph nodes ; Spleen ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To examine the development of the postnatal immune response to thymus-independent type-1 (TI-type 1) and TI type-2 antigens, respectively, trinitrophenyl-lipopolysaccharide (TNP-LPS) or TNP-Ficoll was injected subcutaneously into the hind footpads of young rats of various ages. After 5 days the popliteal lymph nodes (PLNs) were removed and the localization pattern of specific anti-TNP antibody-containing cells was studied. The first specific antibody-containing cells elicited in rats by TNP-LPS appeared in animals at day 19 after birth. The results suggest that the development of these cells from lymphocyte to plasma cell occurs while they migrate from cortex to medulla. An unexpected finding was the low response to TNP-Ficoll in PLN; from 6 weeks after birth only very few specific antibody-containing cells were found. However, in the spleen numerous anti-TNP antibody-containing cells were found in the periarteriolar lymphocyte sheaths. To test the exclusive role of the spleen in the appearance of anti-TNP antibody-containing cells in lymph node after subcutaneous administration of TNP-Ficoll, the experiment was repeated in rats that had been splenectomized. Evidence from these experiments suggests that the spleen plays a major role in the appearance of the above-mentioned cells in lymph nodes.
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  • 110
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    Cell & tissue research 247 (1987), S. 179-185 
    ISSN: 1432-0878
    Keywords: Image analysis ; Morphometry ; Peroxisomes ; Catalase ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The feasibility of the application of a television-based image analyzer, the Texture Analysis System (TAS, Leitz Wetzlar, FRG) in conjunction with a light microscope for morphometric studies of hepatic peroxisomes has been investigated. Rat liver peroxisomes were stained with the alkaline-DAB method for localization of catalase and semi-thin (0.25 and 1 μm) sections of plastic-embedded material were examined under an oil immersion objective. The TAS detected the peroxisomal profiles selectively and determined their morphometric parameters automatically. The same parameters were obtained also by morphometric analysis of electron micrographs from the same material. The volume density of peroxisomes determined by TAS in semithin sections of normal liver, after correction for section thickness, is quite close to the corresponding value obtained by morphometry of electron micrographs. The difference is approximately 20%. In animals treated with the hypolipidemic drug bezafibrate, which causes proliferation of peroxisomes, TAS detected readily the increase in volume density of peroxisomes in semithin sections. In comparison with electron microscopy, however, the light-microscopic approach seems to underestimate the proliferation. The lower resolution of the light microscope and overlapping of neighbouring particles in relatively thick sections used for lightmicroscopic analysis may account for the differences. The present study has demonstrated the usefulness of automatic image analysis in conjunction with selective cytochemical staining of peroxisomes for morphometry of this organelle in rat liver. The light-microscopic approach is not only faster but is also extremely economical by obviating the use of an electron microscope.
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  • 111
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Proteinase inhibitor ; Feedback regulation ; Cholecystokinin ; Fine structure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Application of a single dose of a new type of proteinase inhibitor camostate (FOY-305) via orogastric tube was used in rats to study the dose-response relationship of resulting pancreatic stimulation. Doses up to 10 mg/ kg failed to elicit any response, while significant decrease in enzyme content and increase in serum CCK-levels were observed with doses ranging from 25 to 400 mg/kg. A single dose of 100 mg/kg was selected for a time-sequence analysis, which revealed a 60 to 70% depletion of enzyme stores persisting over 6 h and reverting to control levels by 12 h. Peak increases in serum CCK-levels (15-fold above the elevation observed after regular food intake) were found after 30 min and persisted as an 8-to 10-fold elevation for at least 3 h, then declined to control levels by 9 h. This prolonged endogenous hormone release and resulting pancreatic stimulation were also verified in a separate group of animals in which volume, protein, and enzyme output were measured after cannulation of the pancreatic duct. While volume secretion was not altered by feeding a single dose of 100 mg/kg FOY-305, protein and enzyme output increased 2-to 3-fold over a period of 7 h. Fine-structural analysis of the pancreas demonstrated efficient depletion of zymogen granules from acinar cells with all doses between 50 and 400 mg/kg, accompanied by the appearance of membrane material in the acinar lumina at 3 and 6 h. The same transient increase in the number of lysosomal bodies predominantly containing mitochondria with all doses above 50 mg/kg was interpreted as increased organelle turnover due to persisting hormonal stimulation.
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  • 112
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Caerulein ; DNA synthesis ; Mitotic activity ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous studies from our laboratory indicate that the adaptive response of the exocrine pancreas of the rat to prolonged stimulation with optimal doses of caerulein (0.25 μg × kg-1 × h-1) follows a characteristic time course in which each step in the secretory pathway is activated. The immediate response is the depletion of zymogen-granule stores followed by coordinate and anticoordinate changes in individual rates of (pro-)enzyme synthesis after a lag period of 2 h. The sum of such changes leads to an increase in total rate of protein synthesis by 3 h which is combined with acceleration of intracellular transport packaging and granule discharge. In the present study the time course of DNA synthesis and the labeling index of five populations of pancreatic cells have been analyzed after caerulein stimulation for periods ranging from 6 to 72 h, using in vivo labeling with 1 μCi/g 3H-thymidine 1 h prior to sacrifice of the animals. DNA synthesis did not change during the initial 18 h in spite of persistent stimulation indicated by a 80% reduction of enzyme content. Following this lag period a sharp rise in DNA synthesis 20- to 25-fold above control levels was observed, which decreased by 48 h to reach control levels by 72 h. Increase in DNA synthesis was most pronounced in animals with lowest enzyme content in the pancreas. From the five cell populations studied by autoradiography interlobular duct cells and islet cells had no significant increase in labeling index at any time of stimulation. Acinar cells, intralobular duct cells and interstitial cells showed a marked increase in labeling index after a latent period of 18 h with peak values at 36 h 30 to 50 times higher in intralobular duct and acinar cells, respectively, and 4 times higher in interstitial cells. The increased labeling indices in all three cell populations reverted to lower values at 48 h and reached control values by 72 h. The data indicate a phasic and limited growth response of the rat exocrine pancreas to persistent stimulation with acinar cells as the major contributing cell population.
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  • 113
    ISSN: 1432-0878
    Keywords: Dorsal root ganglia ; Neuropeptides ; Coexistence ; Immunohistochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of the indirect immunofluorescence technique the distribution of calcitonin gene-related peptide (CGRP)-like immunoreactivity (LI) has been analyzed in cervical and lumbar dorsal root ganglia of untreated and colchicine-treated rats. In addition, lumbar ganglia were examined 2 weeks after transection of the sciatic nerve. The occurrence of CGRP-positive cells in relation to ganglion cells containing substance P-, somatostatin-, galanin-, cholecystokinin (CCK)-, and vasoactive intestinal polypeptide (VIP)/peptide histidine isoleucin (PHI)-LI has been evaluated on consecutive sections as well as using elution-restaining and double-staining techniques. CGRP-LI was observed in many ganglion cells of all sizes ranging in diameter from 15 μm to 65 μm. Thus, this peptide occurs also in the large primary sensory neurons. In contrast to the sensory peptides described to date, CGRP-positive cells constituted up to 50% of all and 70% of the medium-sized neurons, thus being the most frequently occurring peptide in sensory neurons so far encountered. Subpulations of CGRP-positive neurons were shown to contain substance P-, somatostatin-, or galanin-LI and some CGRP-positive neurons contained both substance P- and galanin-LI. In fact, most substance P-, somatostatin- and galanin-positive cell bodies were CGRP-immunoreactive. The coexistence analysis further revealed that galanin and substance P often coexisted and that some cells contained both substance P- and somatostatin-LI, whereas no coexistence between galanin and somatostatin has as yet been seen. VIP/PHI-LI was only shown in a few cells in untreated or colchicine-treated rats. However, after transcetion of the sciatic nerve numerous VIP/PHI-positive cells were observed, some of which also contained CGRP-LI. The present results indicate that a CGRP-like peptide is present in a wide range of primary sensory neurons probably not related to specific sensory modalities. Often this peptide coexists with other biologically active peptides. Taken together these findings suggest that CGRP may have a generalized function.
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  • 114
    ISSN: 1432-0878
    Keywords: Cerebral aqueduct ; Ependyma ; Cystic ependymal cells ; Ectopic ependymal cells ; Ependymal ridge ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The wall of the cerebral aqueduct was examined in 20 male rats at the light- and electron-microscopic levels. Disorder in ciliary orientation was occasionally seen in ordinary ependymal cells. Ependymal cells possessing intracellular cysts of 5 to 30 urn in diameter were observed within and beneath the aqueductal ependyma in all animals examined. Light-microscopic reconstruction from serial, 10-μm thick frontal sections revealed an extensive distribution of cystic ependymal cells (CECs), especially along the ependymal ridges in the rostral half of the aqueduct, and along the dorsal region of the aqueductal lining in the caudal half. Both cystic and surface membranes of CECs bore microvilli and cilia. Ectopic ependymal cells (EECs) characterized by densely packed microvilli, well-developed intermediate junctions and cilia, but without cysts, were situated in the subependymal region adjacent to a CEC or another EEC. The ependymal ridges were long, narrow and sporadically stratified ependymal linings extending rostrocaudally and bilaterally along the aqueductal surface. Tanycyte-like cells filled the surface region of the ridge, and CECs and EECs were frequently seen in the core. Intraventricularly injected microperoxidase was detected among densely packed microvilli but not in the cystic lumina of CECs, indicating that EECs and CECs are distinct entities.
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  • 115
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    Cell & tissue research 248 (1987), S. 491-497 
    ISSN: 1432-0878
    Keywords: Intestine, small ; Collagen ; Submucosa ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Whole-mount preparations of the submucosa were made from the small intestine of rats, guinea-pigs, rabbits and sheep. In the distended intestine the collagen fibres ran straight and approximately parallel to the serosal surface. They formed a characteristic lattice, with two arrays of fibres running diagonally in a clockwise and an anticlockwise direction, and making an angle of 50°–55° with the longitudinal axis of the intestine. This collagenfibre lattice was flexible and changed with the movements of the intestinal wall; when the radial distension predominated, the angle between collagen fibres of the submucosa and longitudinal axis of the intestine increased to 60°–65°, and when the longitudinal distension predominated the angle decreased to about 30°.
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  • 116
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    Cell & tissue research 248 (1987), S. 505-510 
    ISSN: 1432-0878
    Keywords: Textile blood-vessel prosthesis ; Aorta ; Neointima formation ; Dog ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The formation of a neo-intima in textile prostheses implanted in the rat and dog aorta was studied by means of light- and scanning electron microscopy. Two independent cellular layers (the superficial and deep ingrowth layers) developed on the free surface and under the fibrin layer initially deposited on the inner surface of the prostheses. The superficial ingrowth layer invades the prosthesis from both the proximal and distal aortic stumps and extends over the primary fibrin layer, or replaces it. This layer consists mainly of smooth muscle cells of the triangular aortic type covered by endothelial-like cells. The deep ingrowth layer originates from cellular elements of the prosthetic bed. Fibroblasts, myofibroblasts and spindle-shaped smooth muscle cells invade the fibrin layer through the interstices of the fabric structure of the prosthesis. Precursors of endothelial cells, however, are absent from this population. The superficial and the deep ingrowth layers may become joined by progressive replacement of the fibrin layer, but remain distinguishable because of their different cellular components. When a continuous cellular layer is established on the inner surface of the prosthesis, and this is then covered by endothelial-like cells, the neo-intima formed remains stable during long-term studies.
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  • 117
    ISSN: 1432-0878
    Keywords: Gonadoliberin ; Pituitary gland, pars anterior ; Gonadotrophs ; Autoradiography ; Cryoultramicrotomy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To identify anterior pituitary cell types containing GnRH binding sites and to study the internalization process of this peptide by target cells under physiological conditions, autoradiography was performed on rat anterior pituitaries removed at specific time intervals (2–60 min) after intravenous injection of mono-radioiodinated 125I-GnRH into intact males. At electron-microscopic level, gonadotrophs and lactotrophs appeared to contain silver grains. Concomitant administration of an excess of unlabeled GnRH with the radioiodinated hormone prevented this localization indicating the specificity of the reaction. The time-course study in gonadotrophs showed that 2 min after injection silver grains could be found over the plasma membrane, secretory granules and nuclear membrane. Similar results were observed 5 and 15 min after injection. Extensive label was observed over the nucleus and nuclear membrane 15 to 60 min after injection. The injection of a radioiodinated GnRH agonist [D-Trp6, Pro9 (Net), DesGly10]-GnRH produced comparable results. In contrast, the injection of 125I-[D-pGlu1, D-Phe2, Trp3,6]-GnRH, an antagonist of GnRH, produced positive labeling only at the plasma membrane without internalization. These results indicate that, after binding with receptors on the plasma membrane, GnRH is rapidly internalized, accumulating in secretory granules, and localizing over the nuclear membrane and later, in the nucleus. Association of radioactivity with secretory granules could be related to a specific action of GnRH at this level or to receptor recycling, and presence of label in the nucleus may be related to stimulation of neosynthesis of LH and GnRH receptors.
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  • 118
    ISSN: 1432-0878
    Keywords: Microtubules ; Terminal web ; Small intestine ; Electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The terminal web (TW) region of mature absorptive cells in the small intestine of the rat contains an elaborate cytoskeleton which supports the apical microvillus membrane. In studies regarding the structural organization of the cytoskeleton and associated proteins in the small intestine, microtubules have not been mentioned as components of the TW. By transmission electron microscopy of conventional resin-embedded sections of rat small intestine, we observe many microtubule profiles in the TW of mature absorptive cells. These microtubules are found in various orientations, although most course parallel to the long axis of the cell, and many microtubule profiles are seen in close association with smooth-surfaced vesicles.
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  • 119
    ISSN: 1432-0878
    Keywords: Exocrine pancreas ; Proteinase inhibitor ; Cholecystokinin ; Protein synthesis ; Enzyme synthesis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Oral application of a single dose of a new synthetic proteinase inhibitor Camostate (Foy-305) in male Wistar rats was carried out together with studies of in vitro amino acid incorporation followed by separation of proteins by two-dimensional gel electrophoresis. The aim of this experiment was to analyze changes produced by the inhibitor in total protein and individual enzyme biosynthesis. Administration of 100 mg/kg Foy-305 resulted in significant inhibition of total pancreatic protein synthesis, without changes in fractional rates for individual enzymes. 50 mg/kg Foy-305 induced a 10-fold elevation of cholecystokinin (CCK) levels in serum; this persisted for 3 h and led to a significant increase in the total rate of protein synthesis with peak values at 6 and 9 h (78% and 84% above control levels, respectively), returning to control by 15h. Changes in fractional rates of synthesis occurred with a latency of 6 h and were restricted to amylase and the anionic form of trypsinogen and chymotrypsinogen. Amylase biosynthesis decreased by about 40% from control levels at 9 h to return to control levels by 15 h. Increased synthesis of trypsinogen and chymotrypsinogen was observed; this was also phasic. The results show similar enzyme-specific regulation as previously described for exogenous CCK stimulation and for the adaptation of the pancreas to diets enriched in protein. They demonstrate the effectiveness of pulsatory endogenous hormone release in the regulation of protein synthesis.
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  • 120
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    Cell & tissue research 249 (1987), S. 215-220 
    ISSN: 1432-0878
    Keywords: X-ray microanalysis ; Thymocyte ; Mitosis ; Sodium ; Potassium ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Elemental concentrations of rat thymocytes in vivo were studied by X-ray microanalysis of freeze-dried sections. Cells from different regions, the subcapsular zone, the cortex and the medulla were studied in thymic tissue from a number of animals. Generally thymocytes situated in the medulla had higher concentrations of K compared to those in the subcapsular zone. The concentration of Na in the nucleus was constant in the medulla in all animals but some variation in this element was seen between animals in the subcapsular zone. The distribution of K/Na ratio in individual thymocytes was different in each region of the thymus. Cells with low K/Na ratio (〈5) were predominant in the subcapsular zone, whereas cells with higher values for K/Na ratio were found in the cortex and medulla. The subcapsular zone is the region where mitotic cells are mostly situated. The finding of thymocytes with higher concentrations of Na and low K/Na ratios in this region is in accord with in vitro studies on thymocyte stimulation.
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  • 121
    ISSN: 1432-0878
    Keywords: Liver ; LDL-gold conjugates ; Albumin-gold conjugates ; Endocytosis ; B, E receptor ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To elucidate the participation of fetal rat liver cells in the receptor-mediated internalization of low-density lipoproteins (LDL), rat fetuses were injected with either LDL-gold or albumin-gold conjugates. The degree of binding and uptake of LDL-gold and albumin-gold by parenchymal and sinusoidal cells of the fetal rat liver differs markedly. Endothelial cells exhibit low LDL-gold uptake. In contrast, parenchymal cells internalize LDL-gold more actively (45 ± 8 LDL conjugates/100 μm2 cytoplasm within 60 min). Kupffer cells exceed this value by a factor of 20. The uptake of albumin-gold by endothelial and Kupffer cells is high, whereas it is extremely low in parenchymal cells. Estradiol pretreatment causes a significant doubling (p〈0.05) of the LDL-gold particle density/100 μm2 cytoplasm both in parenchymal and Kupffer cells, whereas estradiol has no effect on the albumin uptake. The results strongly indicate that LDL uptake by parenchymal and Kupffer cells in the fetal rat liver is mediated by estrogen-inducible receptors, which may correspond to B, E receptors in the adult liver.
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  • 122
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    Cell & tissue research 249 (1987), S. 235-239 
    ISSN: 1432-0878
    Keywords: Satellite nucleoli ; Normal, stimulated and malignant hepatocytes ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study was undertaken to provide information on the presence and frequency of satellite nucleoli in cells with increased nucleolar biosynthetic activity. The number of hepatocytes containing satellite nucleoli was analyzed in rat liver, regenerating liver 18 h after partial hepatectomy and in Novikoff hepatoma ascites cells. In comparison with hepatocytes of normal liver, the number of both stimulated hepatocytes and malignant hepatoma cells containing satellite nucleoli was significantly reduced. The results also indicated that whereas most satellite nucleoli contain protein C23, a smaller percentage contain protein B23.
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  • 123
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    Cell & tissue research 249 (1987), S. 367-377 
    ISSN: 1432-0878
    Keywords: Ethane dimethanesulphonate ; Leydig cells ; Destruction ; Regeneration ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ethane dimethanesulphonate (EDS) was used as a specific cytotoxin to eliminate the Leydig cell population of the adult rat testis. Ultrastructural, morphometric and serum gonadotrophin and testosterone analysis was used to study the response of the intertubular tissue of the testis from 1 day to 10 weeks after EDS treatment. In control animals, the testis contained approximately 28 million Leydig cells and 8 million macrophages. Three to seven days after EDS treatment, Leydig cells were absent and serum testosterone was undetectable. Macrophage numbers increased three-fold by 3 days and returned to pretreatment values thereafter. At 2 and 3 weeks post-EDS, foetal-type Leydig cells (∼1–2 million per testis) appeared in proximity to perivascular and peritubular tissues, a feature also observed at 4 weeks when numerous such cells (∼15 million per testis) formed prominent clusters in perivascular and peritubular locations. Between 6 and 10 weeks after EDS treatment, the foetal-type Leydig cells were transformed morphologically into adult-type Leydig cells, they occupied central intertubular positions and their numbers were restored to pretreatment values. Regeneration of Leydig cells was reflected by elevated serum testosterone levels which returned towards the normal range. The results demonstrate the regenerative capacity of the testicular intertubular tissue and indicate a dual site of origin of Leydig cells which initially resemble foetal-type Leydig cells prior to establishing the adult-type Leydig cell population. The morphological pattern of Leydig cell regeneration suggests that in addition to gonadotrophic stimulation, local testicular factors from the seminiferous tubules may stimulate Leydig cell growth.
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  • 124
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    Cell & tissue research 248 (1987), S. 103-110 
    ISSN: 1432-0878
    Keywords: Secretory ameloblast ; Ca++-ATPase ; Na+-K+-ATPase ; Ultracytochemistry ; Calmodulin blocker, trifluoperazine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of the calmodulin blocker, trifluoperazine (TEP), on membrane-bound Ca++ -ATPase, Na+ -K+ -ATPase (EC 3.6.1.3.) and the ultrastructure of the enamel organ were investigated in the lower incisors of normal and TFP-injected rats. The rats, of about 100 g body weight, were given either 0.2 ml physiological saline or 100 μg TFP dissolved in 0.2 ml physiological saline through a jugular vein and fixed by transcardiac perfusion with a formaldehyde-glutaraldehyde mixture at 1 and 2 h after TFP administration. Non-decalcified sections of the enamel organ less than 50 μm in thickness, prepared from dissected lower incisors, were processed for the ultracytochemical demonstration of Ca++-ATPase and Na+-K+ -ATPase by the one-step lead method at alkaline pH. In control saline-injected animals the most intense enzymatic reaction of Ca++-ATPase was demonstrated along the plasma membranes of the entire cell surfaces of secretory ameloblasts. Moderate enzymatic reaction was also observed in the plasma membranes of the cells of stratum intermedium and papillary layer. Reaction precipitates of Na+-K+-ATPase activity were localized clearly along the plasma membranes of only the cells of stratum intermedium and papillary layer. The most drastic effect of TFP was a marked disappearance of enzymatic reaction of Ca++-ATPase from the plasma membranes of secretory ameloblasts, except for a weak persistent reaction in the basolateral cell surfaces of the infranuclear region facing the stratum intermedium. The cells of stratum intermedium and papillary layer, however, continued to react for Ca++-ATPase even after TFP treatment. Similarly, Na+-K+-ATPase activity in these cells was not inhibited by TFP administration. Ultrastructural examination of secretory ameloblasts revealed that administration of TFP caused no considerable cytological changes and did not act as a cytotoxic agent. These results suggest that secretory ameloblasts may have an active Ca++ transport system, which is modulated by an endogenous calmodulin.
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  • 125
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    Cell & tissue research 248 (1987), S. 79-88 
    ISSN: 1432-0878
    Keywords: Bone resorption ; Implantation ; Osteoclasts ; Osteopetrosis ; Mononuclear phagocytes ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Faulty osteoclasts, characteristic of the incisors-absent (ia) rat mutation of osteopetrosis, cause a resorptive defect which results in the persistence of immature, highly mineralized bone matrix. We implanted osteopetrotic bone subcutaneously into normal andia rats to determine ifia bone could induce functionally active and morphologically identifiable osteoclasts at the implant surface. Assays of45Ca released from the preparations showed that normal andia recipients were capable of equivalent cell-mediated release of Ca over a 2-week implant period, indicating that theia resorptive defect was not reproduced at the subcutaneous site. Freeze-thawed osteopetrotic bone released twice as much45Ca as normal bone. This difference was eliminated by collagenase treatment. Cellular profiles were similar in both normal andia animals regardless of the implant preparation. At 3 days after implantation, both bone and suture were surrounded by mononuclear cells. By 14 days, multinucleated cells appeared at the implant surfaces. Morphological comparison of implant-induced multinucleated cells and tibial osteoclasts indicated that bone-elicited multinucleated cells lacked the ruffled borders characteristic of normal osteoclasts or the extensive clear zones typical ofia osteoclasts, but more closely resembled suture-induced macrophage-polykaryons. We conclude that ectopically implantedia bone as compared to normal bone elicits a different functional response from structurally similar cell populations. Bone-elicited multinucleated cells could not be classified as active osteoclasts despite evidence of release of45Ca. Release of labeled Ca was probably due to the action of mononuclear phagocytes and macrophage-polykaryons rather than to osteoclastic resorption.
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  • 126
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    Cell & tissue research 248 (1987), S. 119-123 
    ISSN: 1432-0878
    Keywords: Odontoblasts ; Protein synthesis ; Denervation ; Neural regulation ; Wound healing ; Autoradiography ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Odontoblasts respond to occlusal trauma by increased elaboration of a matrix which is subsequently calcified to form reparative dentin. The purpose of the present study was to analyze quantitatively and compare the ability of odontoblasts to synthesize collagen after wounding in rats with an intact innervation (baseline) and in rats with sensory (inferior alveolar nerve, IAN) and/or sympathetic (superior cervical ganglion, SCG) surgical denervation. Surgery was performed 7 days prior to wounding. All rats had 1 mm of enamel and dentin removed from the occlusal surface of the first mandibular molar (resected side) with the contralateral tooth serving as a control. Rats were killed 1 h after injection with3H-proline on days 0, 5, 10 or 15 after wounding, and mandibles were removed and processed for autoradiography. Grain counts were performed over odontoblasts throughout the pulp horns for each time period and for control and experimental molars in intact (baseline) and denervated groups. When compared to the control baseline, the experimental baseline data showed increased3-proline uptake throughout the study with a peak at 5 days. When compared to the baseline data, IAN and SCG results demonstrated a delay or attenuation of the protein synthetic response. The results indicate that the sensory and sympathetic neural components may regulate odontoblastic response to wounding.
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  • 127
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Alkaline phosphatase ; Small intestine ; Colchicine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The purpose of this study was to investigate the effect of colchicine and vinblastine on the localization of alkaline phosphatase (AlPase) in rat duodenum in relation to structural changes. AlPase was localized on the membranes of rough endoplasmic reticulum, Golgi stacks, cytoplasmic vesicles, microvilli, on lateral plasma membranes, and in some lysosomes of the duodenal epithelial cells of rats treated with either lumicolchicine or 0.9% NaCl alone. Microvilli were most intensely stained, and AlPase-positive Golgi stacks were regularly distributed in the supranuclear regions. After colchicine treatment, microvilli were shortened and the staining intensity became weaker, whereas basal as well as lateral plasma membranes showed stronger staining. The AlPase-positive microvilli appeared not only on the luminal surfaces, but also on the baso-lateral plasma membranes and even on the surfaces of characteristic intracytoplasmic cysts. Golgi stacks became smaller and their distribution became less localized, and the staining intensity of the Golgi stacks became weaker. AlPase localization in rats treated with vinblastine was almost identical with that of rats treated with colchicine. Thus, colchicine and vinblastine appeared to have elicited a disorientation of intracellular transport of intestinal AlPase by inhibiting microtubule organization.
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  • 128
    ISSN: 1432-0878
    Keywords: Insulin-like growth factor I ; Somatomedin C ; Arterial injury ; Endothelial regeneration ; Intimai thickening ; Macrophages ; Smooth muscle cells ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the present study the expression of insulin-like growth factor I (IGF-I; somatomedin C) immunoreactivity was examined in endothelial cells during repair after injury to the intima in the femoral artery of adult rats. Two types of injury were examined: (1) endothelial denudation induced by the use of a catheter, and (2) vessel compression by short-term ligation. In untreated rats, arterial endothelial cells showed no or, only infrequently, low IGF-I immunoreactivity in their cytoplasm. Endothelial cells at the border to the denuded area showed increased IGF-I immunoreactivity one day after injury to the intima of the femoral artery. Thrombocytes and fibrin deposits as well as vital endothelial cells, covered by clots, were immunonegative. The maximal intensity of IGF-I immunoreactivity was reached within 3 days after insult. The IGF-I immunoreactivity in the endothelial cells remained elevated for at least 4 weeks, compared to the controls. Intimai thickenings appeared within a week after injury and many cells in these thickenings showed intense IGF-I immunoreactivity as did the covering endothelial cells. Smooth muscle cells in the media were generally immunonegative during control conditions and after endothelial denudation. Spontaneously hypertensive rats (SHR) showed, similarly to their matched controls (WKY), approximately the same patterns of IGF-I immunoreactivity in their endothelial cells both under normal conditions and after injury. It is concluded that IGF-I is likely to be involved in the repair of the intima in injured arteries.
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  • 129
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Cyanoketone ; Corticosterone ; Morphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of a 7-day administration of cyanoketone (CKT) on the adrenal zona fasciculata were examined in “normal” and dexamethasone/ACTH-infused rats. The drug caused a 48–56% decrease in the blood concentration of corticosterone, coupled with a 53–58% lowering in the activity of 3β-hydroxysteroid dehydrogenase (3βHSD), in both groups of animals. In the “normal” rats, CKT induced a significant hypertrophy of the zona fasciculata and its parenchymal cells, due to an increase in volume of the mitochondrial compartment and to proliferation of the smooth endoplasmic reticulum (SER), as well as a notable rise in both the volume of the lipid-droplet compartment and the intracellular concentration of total cholesterol. The activity of 11β-hydroxylase was conspicuously enhanced. All these responses of zona fasciculata cells to CKT did not occur in dexamethasone/ACTH-treated animals, and therefore they were considered to be non-specific and mediated by the augmented secretion of ACTH following lowering of the corticosterone level in the blood. In the dexamethasone/ACTH-infused rats, the only morphological change induced by CKT was a significant decrease in the surface area per cell and surface density of the SER, which was interpreted as the morphological counterpart (not masked by the increased level of circulating ACTH) of the drug-induced inhibition of the microsomal 3βHSD.
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  • 130
    ISSN: 1432-0878
    Keywords: Alpha-bungarotoxin ; Autoradiography ; Cholinergic receptors ; Electron microscopy ; Suprachiasmatic nucleus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of (125I) alpha bungarotoxin (α-BTX) binding sites in the suprachiasmatic nucleus (SCN) of the adult female rat was examined by electron-microscopic autoradiography. The ultrastructural distribution of silver grains was analysed by line source, direct point count, and 50% probability circle methods. Real grain distribution was significantly different from that of randomly generated hypothetical grains. Line source analysis demonstrated two populations of sources: one associated with membranes, and one inside neuronal structures. Probability circle analysis of shared grains indicated that membrane-bound-radioactive sources were mainly asssociated with axo-dendritic appositions. Only a small proportion of labeled neuronal interfaces exhibited synaptic differentiations in the plane of section. However, the compartment containing synaptic terminals was the most enriched when comparing real to hypothetical grains. Probability circle analysis of exclusive grains demonstrated that sources that were not associated with neuronal plasma membranes were likely to be within nerve cell bodies and dendrites. It is concluded that the majority of specifically labeled α-BTX binding sites in the SCN is membrane bound, and may be associated with axodendritic synaptic transmission. The presence of a significant proportion of the label in the soma and dendrites of suprachiasmatic neurons 24 h after ventricular infusion suggests that some of the labeled binding sites (junctional or nonjunctional) may be internalized within these two compartments.
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  • 131
    ISSN: 1432-0878
    Keywords: Salivary glands ; Chondroitin sulfate ; Secretory granules ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The presence of chondroitin sulfate in secretory granules of the rat parotid gland and its saliva was revealed by radioactive sulfate incorporation, followed by isolation and partial characterization of the sulfated species contained within the granules and in the parotid saliva. 35SO 4 = was incorporated into chromatographically identical macromolecular material both in vitro, in a gland-slice system followed by isolation of granule contents, and in vivo as measured in the pure parotid secretion following intravenous administration of 35SO 4 = . The majority of the 35SO 4 = label appeared in a peak in the region where the family of acidic proline-rich proteins elute from a DEAE-Sephadex A-50 column. Papain digestion freed the sulfated material from the bulk of protein present in this peak, leaving sulfate-labelled material that chromatographed on Sepharose CL6B as a single peak corresponding to a molecular weight of 13000 daltons. The ratio of uronic acid to amino sugar in this sulfated peak was 0.56. The sulfated material was susceptible to degradation by chondroitinase AC. The presence of this chondroitin sulfate in secretory granules and saliva is consistent with previous suggestions that sulfated polyanions may play a role in formation and maturation of secretory granules.
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  • 132
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    Cell & tissue research 250 (1987), S. 257-266 
    ISSN: 1432-0878
    Keywords: Choriocapillaris ; Endocytosis ; Endothelium ; Eye ; Glycosaminoglycan ; Heparin ; Receptors ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Heparin-gold probes were used to localize regions of heparin binding on the luminal surface of the diaphragmed-fenestrated endothelium of the rat choriocapillaris. Structures of endothelial cells were unlabeled when rats were kept on ice and perfused with solutions at 4° C. When the heparin-gold quantity was doubled, only a few heparin-gold particles marked some diaphragms spanning fenestrae, vesicles and channels, parajunctional regions of the plasmalemma, and coated pits. With solutions at 4° C, but the animals kept at room temperature, all of these structures in the endothelial cells were labeled. This binding was not altered by the perfusion of low levels of unlabeled heparin, but was eliminated following high levels of unlabeled heparin, and by digestion with trypsin and pronase. At 37° C, heparin was localized to the above structures and, in addition, was internalized into coated vesicles, endosomes, and multivesicular bodies, but not other types of lysosomes. Some particles were found in tubules adjacent to the Golgi stacks. Heparin-gold was also transported to the abluminal front of the endothelium by vesicles. A desulfated, non-anticoagulant, fraction of heparin bound to gold was localized to the endothelium in the same manner. These results demonstrate receptors for heparin on the surface of a fenestrated endothelium. Furthermore, they show the pathway of endocytosis and transport of heparin. The possible roles of heparin in the function of the endothelium is discussed.
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  • 133
    ISSN: 1432-0878
    Keywords: Galanin ; Iris ; Choroid membrane ; Immunohistochemistry ; Trigeminal ganglion ; Superior cervical ganglion ; Calcitonin gene-related peptide ; Capsaicin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The iris and choroid membrane of the adult rat contain nerve fibers expressing immunoreactivity to the neuropeptide galanin. The density and distribution of galanin-positive nerve fibers varied from iris to iris and, particularly, among animals. Smooth, non-terminal axons were seen running in nerve bundles consisting of otherwise negative fibers. From the choroid membrane these bundles reached the iris via the ciliary body. Axons were frequently seen to branch giving rise to a sparse system of varicose, single fibers in the dilator plate and sphincter area. Galanin-positive fibers were sometimes also seen outlining blood vessels. Capsaicin, in a dose that causes permanent depletion of substance P- and cholecystokinin-immunoreactive fibers in the iris, caused no change in amount of galanin-positive fibers. Removal of the superior cervical ganglion caused a rapid and pronounced increase in the number of galanin-immunoreactive nerve fibers. Similarly, removal of the ciliary ganglion appeared to increase galanin immunoreactivity, while removal of the pterygopalatine ganglion was less effective. Lesioning of the trigeminal ganglion caused a disappearance of galanin immunoreactivity. The sympathetectomy-induced increase was counteracted by capsaicin. Galanin-positive nerve cell bodies were present in both the superior cervical and the trigeminal ganglia. In the superior cervical ganglion, immunoreactive galanin did not seem to coexist with neuropeptide Y-positive cells; in the trigeminal ganglion, some galanin-positive cells also contained calcitonin gene-related peptide (CGRP) immunoreactivity, while most cells did not. In the iris, double-staining suggested that CGRP and galanin immunoreactivities were contained in different fiber populations. We conclude that the rat iris and choroid membrane contain a sparse plexus of nerve fibers expressing galanin-like immunoreactivity. It is suggested that these fibers are derived from the trigeminal ganglion. The iris is able to respond with a pronounced increase in number of galanin-immunoreactive nerve fibers to certain denervation procedures.
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  • 134
    ISSN: 1432-0878
    Keywords: Polypeptide 7B2 ; Adenohypophysis ; Secretion ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A highly conserved polypeptide termed “7B2”, isolated from human and porcine pituitaries, has been reported by immunoreactivity to be distributed in various organs. However, the highest concentration has been found in the pituitary as demonstrated by a specific radioimmunoassay. In order to determine the type of cells within the pituitary that contain 7B2 and to analyse its intracellular localization, specific immunocytochemistry techniques (unlabeled antibody, peroxidase-antiperoxidase) were used both for light and electron microscopy. Immunocytochemistry of both expiants and monolayer-cell cultures of the adenohypophysis was studied. Immunoreactivity to 7B2 has been found in 21.9% of the total number of cells. After simultaneous staining of serial sections with appropriate antibodies, 7B2 was found to be colocalized with β-LH/β-FSH in gonadotrophs and with β-TSH in thyrotrophs. In situ immunocytochemistry at the electron-microscopic level showed that immunoreactive 7B2 is compartmentalized within secretory granules. The small (130 to 250 nm) but not the large granules (400 to 700 nm) were labeled in gonadotroph-like cells and small granules (90 to 150 nm) were also labeled in thyrothrophlike cells. Study of the gonadotrophs in cell culture after Zamboni's fixation revealed weak to moderate immunoreaction in rough endoplasmic reticulum. The current findings as well as previous data indicate that 7B2 is synthesized, stored and possibly released from the adenohypophysis similarly to many other secretory products.
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  • 135
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    Cell & tissue research 250 (1987), S. 215-220 
    ISSN: 1432-0878
    Keywords: Gossypol ; Ovarian follicle ; Granulosa cells ; Lysosomes ; Smooth endoplasmic reticulum ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural changes were observed in the oocytes and granulosa cells of the ovarian follicles of sexually mature female rats treated daily for 60 days with gossypol acetic acid at a dosage known to induce sterility in male rats (20 mg/kg body weight, orally). The changes included an increased lysosomal population in the ooplasm and a reduced, poorly developed smooth endoplasmic reticulum in the granulosa cells. The latter changes suggest a suppression of the secretory activity of the granulosa cells.
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  • 136
    ISSN: 1615-6102
    Keywords: Silver staining ; Nucleolus ; Quantitative analysis ; Pineal gland ; Rat ; Oestrous cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Morphology, distribution and number of different argyrophilic aggregates appearing in pinealocyte nuclei have been studied during the rat oestrous cycle. Using the Ag-NOR reaction we have found two types of argyrophilic aggregates in pinealocyte nuclei. One of them, corresponding to the fibrillar centres and the surrounding fibrillar component, appears in the nucleoli. The silver grains are more loosely packed in the dense fibrillar component than in the fibrillar centres. A decrease in the number of these nucleolar argyrophilic aggregates was obtained at oestrous. A second type of silver grain aggregate was observed in the pinealocyte nucleoplasm. We call them “Ag-granule clusters” because they are similar to the interchromatin granule clusters and constitute the only silver deposit forming aggregates apart from the nucleolus. The number of Ag-granule clusters is significantly smaller at oestrous and meta-oestrous than at dioestrous and pro-oestrous.
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  • 137
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    Medical & biological engineering & computing 25 (1987), S. 386-390 
    ISSN: 1741-0444
    Keywords: Central caudal artery ; Electrical impedance plethysmography ; Electromagnetic flowmetry ; Peripheral blood flow ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The impedance Z, the derivative of Z with respect to time t, dZ/dt, and blood flow F have been measured simultaneously in the central caudal artery of a series of anaesthetised rats. The results of the experiments were expressed in the relationship derived from the Windkessel theory where f(t)=adz(t)/dt+βz(t), in which α and β are constants and f(t) and z(t) express the pulsating fractions in the blood flow F and the impedance Z, respectively. The correlation between the peak-to-peak values of f(t) and of adz(t)/dt+βz(t) was 0·96. The correlation between the mean flow fo and the peak-to-peak value of f(t) was 0·74. Furthermore, even when the mean flow was zero, adz(t) still existed. The theoretical model was tested against volume flow measurements made using an electromagnetic flowmeter and was found to correlate well. Theoretical estimates of vessel calibre also accord with measured values.
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  • 138
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    Cellular and molecular life sciences 42 (1986), S. 169-171 
    ISSN: 1420-9071
    Keywords: Rat ; olfactory pathway selective lesions ; plasma corticosterone ; lactacidemia ; adrenal glands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In comparison to control rats, basal plasma corticosterone level and lactacidemia significantly increased in rats submitted to a bilateral lesion of the lateral olfactory tract and/or the anterior branch of the anterior commissure. Only the anterior branch of the anterior commissure induced hyperglycemia; that of the lateral olfactory tract exerted an opposite effect.
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  • 139
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    Cell & tissue research 246 (1986), S. 205-210 
    ISSN: 1432-0878
    Keywords: 1,2-Dimethylhydrazine ; Peptide YY ; Glucagon ; 5-Hydroxytryptamine ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Colorectal adenocarcinomas were induced in male Wistar rats, by weekly subcutaneous administration of 1,2-dimethylhydrazine, classified according to the degree of differentiation and submitted to immunocytochemistry for the peptides cholecystokinin (CCK), gastrin, gastric inhibitory polypeptide (GIP), glucagon, neurotensin, pancreatic polypeptide (PP), peptide YY (PYY), somatostatin and vasoactive intestinal polypeptide (VIP) and the biogenic monoamine 5-hydroxytryptamine. Well- or moderately well-differentiated adenocarcinomas comprised 46% of the tumour population, only 4% were poorly-differentiated adenocarcinomas, and the remaining 50% possessed a mixture of these two morphologies. Glucagon, PYY and 5-hydroxytryptamine immunoreactive cells were frequently observed within well- or moderately well-differentiated tumours and within such regions of tumours possessing a mixed morphological pattern. The tumours contained no cells immunoreactive for any of the peptides not normally located within the colorectum, nor did they contain cells immunoreactive for somatostatin and VIP, although known positive controls did stain. Poorly-differentiated tumours and portions of tumours of mixed type, were consistently negative. 5-hydroxytryptamine was the most frequently located of the three antigens, being detected in 87% of the moderately well-differentiated tumours and 32% of the tumours with mixed morphologies. 11% of moderately well-differentiated tumours possessed 5-hydroxytryptamine positive cells in such profusion that they contributed significantly to the tumour mass. The distribution of glucagon-and PYY-immunoreactive cells was similar, although they occurred with a lower frequency, presumably corresponding to their lower numbers within the normal colorectal mucosa. Additionally, these two peptide immunoreactivities were colocalized in the majority of cells, although some cells contained only one antigen. The immense numbers of cells immunoreactive for peptides and monoamine in a significant proportion of colorectal adenocarcinomas suggests that they have arisen from multipotential endodermal stem cells within the tumours and are not part of the normal epithelial population being engulfed as the tumour grows.
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  • 140
    ISSN: 1432-0878
    Keywords: Arginine vasopressin ; Choroid plexus ; Fluid transport ; Cerebrospinal Fluid ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The experiments described herein use an in vitro preparation of choroid plexus to demonstrate that it is a vasopressin-responsive organ by morphologic criteria. Choroid plexus from rats was incubated for one hour in graded concentrations of arginine vasopressin (AVP). Within physiologic range of molar concentration, incubation in vasopressin induced a decrease in basal and lateral spaces in choroid plexus epithelial cells as well as an increase in number of dark cells. The number of cells with basal spaces decreased significantly from 82.7±9.2 in control tissue to 19±18 in tissue incubated in 10-12 M AVP; similarly, the number with lateral cellular spaces decreased from 20±8.8 to 7.6±2.2 cells in 10-10 M AVP. Dark cells increased in number from 3.8±2.6 in control conditions to 49±4 with 10-9 M vasopressin. These data suggest important effects of arginine vasopressin in cerebrospinal fluid (CSF) on choroid plexus, compatible with enhanced fluid transport across choroid epithelial cells.
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  • 141
    ISSN: 1432-0878
    Keywords: Glucocorticoids ; Cartilage ; Growth ; Histochemistry ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effects of different doses of various steroids on growth, and on costal and epiphyseal chondrocytes, have been studied in prenatal, immature, and adult Long-Evans rats using histochemical techniques, and both light and electron microscopy. Both prenatal and postnatal treatments have been employed. The steroids used were cortisone (CA), betamethasome (BM), and, in the prenatal group only, dexamethasone (DM). Body weight is reduced in all treated rats (except the low dose of CA) by day 17 of gestation, with greater weight reductions occurring in rats receiving the higher dose level of each steroid. In rats treated prenatally or neonatally, and sacrificed postnatally on days 39–43 or days 116–127, body weights, and tibial and tail lengths, are less than in correspondingly aged controls, thus showing a persistence of the effects of treatment. Costal and epiphyseal cartilages in prenatal rats show cellular, synthetic, and ultrastructural alterations induced by treatment with glucocorticoids but the responses are not necessarily comparable. Except for the low dose of DM, the higher doses of each steroid are more effective in inhibiting, or altering, growth and cellular differentiation in the developing fetuses. Surprisingly, a low dose of DM has a more devastating effect on the cells and extracellular matrix of both costal and epiphyseal cartilage, than do higher dose-levels of the various steroids. Low doses of CA and BM are also effective in inhibiting or altering growth and cellular differentiation, but their effectiveness is largely limited to 17 days of gestation. The order of effect of the various doses of the different steroids on fetal cartilage, listed in decreasing order of severity, is as follows: 0.12 DM, 0.24 DM, 0.42 BM, 50 CA, with 25 CA and 0.18 BM being approximately equal and only slightly different from control cartilages. The effect of prenatal or neonatal glucocorticoid treatment on chondrocytes is minimal in the 30–43 day, or 116–127 day, postnatal groups. In immature and adult rats, cortisone affects the chondrocytes more deleteriously than does betamethasone, and a 5.0 mg dose of CA seems to affect chondrocytes, body weight, and tibial and tail lengths more than 0.2 or 7.5 mg doses.
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  • 142
    ISSN: 1432-0878
    Keywords: Microcirculation ; Endothelial vesicles ; Skeletal muscle ; Brain vessels ; Transendothelial transport ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrathin serial sectioning and labeling with tannic acid have demonstrated that most plasmalemmal vesicles of rat vascular endothelial cells are not free, but rather are conjoined in three dimensions to form racemose invaginations from the cell surfaces. To elucidate the distribution of vesicles in these microvascular endothelial cells, we have examined terminal arterioles, capillaries and post-capillary venules of rat skeletal muscle and brain cortex, using tannic acid labeling and stereological methods, and have determined the proportions of free vesicles and the vesicles of luminal and abluminal invaginations, as well as the numerical density of vesicles. In the case of capillaries, regional differences in distribution have also been studied. The ratio of free vesicles is 6–7% and is constant throughout the muscle microvasculature. The distribution (proportions and numerical densities) of vesicles in the brain and muscle microvascular endothelial cells shows regionally distinctive patterns. In rapid-frozen, freeze-substituted endothelial cells, there are almost as many fused vesicles as seen in chemically fixed cells. Therefore, aldehydes do not seem to induce membrane fusion, and the distribution of vesicles seems to be preserved by chemical fixation. The structure and function of plasmalemmal vesicles are discussed.
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  • 143
    ISSN: 1432-0878
    Keywords: Somatostatin (SRIF, GHR-IF) ; Neuropeptides ; Immunohistochemistry ; Thalamus ; Retrograde tracing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of the indirect immunofluorescence technique a small group of large somatostatin-positive neurons is described in the subependymal area of the anterior paraventricular thalamus of the male rat. Retrograde-tracing experiments suggest that they project to areas outside the blood-brain barrier.
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  • 144
    ISSN: 1432-0878
    Keywords: Serotonin-immunoreactive nerve fibers ; Lateral geniculate nucleus ; Immunohistochemistry ; Rat ; Cat ; Monkey (Macaca fuscata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of serotonin-containing nerve fibers in the lateral geniculate nucleus (LGN) of the rat, cat, and monkey (Macaca fuscata) was studied by use of the peroxidase-antiperoxidase method and an antiserum against serotonin. In all three species, the pattern of fibers was denser in the ventral portion of the LGN (LGNv) than in the dorsal nuclear portion (LGNd). In the LGNd of rat, serotonin-immunoreactive fibers were evenly distributed in the form of a dense network, but in cat and monkey there were marked regional differences. Serotonin-immunoreactive elements were most numerous in the C complex and medial interlaminal nucleus of cat, and in the S layer and interlaminar zones of Macaca fuscata.
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  • 145
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; ACTH ; Steroidogenesis ; Aldosterone escape phenomenon ; Stereology ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Short-term ACTH treatment provoked a decrease in volume of the lipid-droplet compartment in rat zona glomerulosa cells, and a rise in plasma and intracellular concentrations of corticosterone and aldosterone. It enhanced activities of 3β-hydroxysteroid dehydrogenase (3βHSD), 11β-hydroxylase (11βOH) and 18-hydroxylase (18OH). Long-term ACTH administration produced a hypertrophy of the zona glomerulosa and its parenchymal cells, a result of the increase in volume of the smooth endoplasmic reticulum and the mitochondrial compartment. The surface area per cell of mitochondrial inner membranes increased; the tubular cristae were transformed into a homogeneous population of vesicles. The plasma and intracellular concentrations of corticosterone further increased, whereas those of aldosterone fell below basal levels (the “aldosterone-escape” phenomenon). The activities of 3βHSD and 11βOH were enhanced, that of 180H decreased. Therefore, ACTH stimulates zona glomerulosa growth and transforms parenchymal elements into zona fasciculata celltypes. Cyanoketone nullified acute ACTH effects on plasma and intracellular concentrations of corticosterone and aldosterone, but did not affect the activities of 11βOH and 18OH. Chronic ACTH treatment produced similar results, although 18OH activity was not suppressed. The mechanism underlying the “aldosterone-escape” phenomenon may thus involve a rise in the intracellular concentration of corticosterone, caused by the enhanced synthesis and activation of 3βHSD and 11βOH.
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  • 146
    ISSN: 1432-0878
    Keywords: Met5-enkephalin-arg6-gly7-leu8 ; Enkephalin ; Gastrointestinal tract ; Immunohistochemistry ; Opioid peptides ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution and characterization of the opioid octapeptide met5-enkephalin-arg6-gly7-leu8 (met5-enk-arg6-gly7-leu8) within the gastrointestinal tract of the rat has been determined by immunohistochemistry and radioimmunoassay by use of a newly developed antibody to met5-enk-arg6-gly7-leu8. With both techniques, met5-enk-arg6-gly7-leu8-immunoreactivity (met5-enk-arg6-gly7-leu8IR) was detected in all regions of the gastrointestinal (GI) tract except the esophagus. The highest concentration of immunoreactive met5-enk-arg6-gly7-leu8 was observed in the colon, while intermediate concentrations were found in the stomach, duodenum, jejunum, and ileum. Immunostained somata were observed chiefly in the myenteric plexus; immunostained processes were present primarily in the myenteric plexus and the circular muscle layer. This distribution pattern is similar to that previously observed with antiserum to met5-enkephalin-arg6-phe7 (met5-enk-arg6phe7). Chromatographic analysis of met5-enk-arg6-gly7leu8-immunoreactive peptides extracted from the GI tract revealed the presence of an immunoreactive peptide of high molecular weight which accounted for approximately three-quarters of met5-enk-arg6-gly7-leu8-IR in both stomach and colon. These findings suggest a role for peptides related to the octapeptide met5-enk-arg6-gly7-leu8 in the regulation of GI function.
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  • 147
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    Cell & tissue research 244 (1986), S. 141-145 
    ISSN: 1432-0878
    Keywords: Adrenal cortex ; Sexual dimorphism ; Gonadectomy ; Stereology ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cytological aspects of sexual dimorphism in the rat adrenal cortex and its relationship to the gonads have been investigated. The adrenal glands of mature female rats were heavier than those of males, and morphometry showed that this was almost exclusively due to conspicuous differences in the volume of cells of the zona fasciculata (ZF) and zona reticularis (ZR). Stereology demonstrated that the volume of the mitochondrial and lipid droplet compartments, as well as the surface area per cell of mitochondrial cristae and smooth endoplasmic reticulum tubules, were markedly higher in the ZF and ZR cells of female animals. Orchiectomy increased and ovariectomy decreased the adrenal weight, by eliciting hypertrophy and atrophy, respectively, of ZF and ZR cells; these effects of gonadectomy were reversed by the appropriate gonadal hormone replacement. It is suggested that the sexual dimorphism of the rat adrenal cortex may depend upon the inhibitory action of testosterone and the stimulatory effect of estradiol on the hypothalamo-hypophyseal-adrenal axis.
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  • 148
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    Cell & tissue research 244 (1986), S. 437-441 
    ISSN: 1432-0878
    Keywords: Mitochondria ; Spermatocytes ; Freeze-fracture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Separation of the two-folded lamina of the mitochondrial cristae occurs in mitochondria of spermatocytes and spermatids. Freeze-fracture exposes large areas of the inner and outer halves of the inner membrane. The surface of the outer half of the inner membrane is concave, with small numbers of intramembranous particles (IMPs). Its distinctive feature is the presence of protruding particles surrounding a pit. On the inner half of the inner membrane, there are large numbers of densely-packed, irregularly-distributed IMPs, among which regular pits are seen. Morphometric analysis and reconstructions suggest that these structures are “channels” in the mitochondrial membrane with an internal diameter of approximately 18 nm. It is uncertain whether such mitochondrial structures are confined to the spermatocyte or whether they may also occur in other cells.
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  • 149
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    Cell & tissue research 244 (1986), S. 467-469 
    ISSN: 1432-0878
    Keywords: Pituitary gland, posterior lobe ; Electron-microscopic autoradiography ; α2-Binding sites ; Clonidine ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Electron-microscopic autoradiography of rat neurohypophyses treated with [3H] clonidine, an α2-agonist, showed that binding apparently occurred preferentially at the neurosecretory endings and blood vessels rather than on the pituicytes. Since it is known that clonidine has a high affinity for plasma proteins, the distribution over the neurosecretory nerve endings would suggest the existence of presynaptic α2-binding sites on neurosecretory neurones, which could indicate a regulatory function for catecholamines in neurohypophysial hormone release.
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  • 150
    ISSN: 1432-0878
    Keywords: GABA ; GABA-transaminase ; Oviduct ; Cilia ; Basal bodies ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution in the rat oviduct of γ-aminobutyric acid and its catabolic enzyme GABA-transaminase was studied by the use of immunocytochemical and enzymehistochemical techniques. At the light-microscopic level, both GABA immunoreactivity and GABA-transaminase enzyme reactivity were found primarily in the tubal epithelium while in the muscle layers of the organ only a faint GABA and GABA-transaminase positive staining could be detected. Electron-microscopic evaluation of the GABA immunoreactivity revealed a heavy labelling of the basal bodies (kinetosomes) and a moderate staining of the cilia. These findings indicate that the role of GABA in the oviduct is not related to neurotransmission but may be related to ciliary functions.
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  • 151
    ISSN: 1432-0878
    Keywords: Urinary bladder ; Discoidal vesicles ; ATP ; Morphometrics ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Movement of asymmetric membrane plaques between the cytoplasm and surface of luminal urothelial cells was investigated during artificially induced contraction and expansion of untreated and ATP-depleted urinary bladders of the rat. Estimations of surface area, volume, and number of discoidal vesicles per unit volume of cytoplasm were determined by morphometric examination of electron micrographs. These values were compared in luminal cells from bladders incubated in control media or in media containing 0.15 mM 2,4-dinitrophenol and 0.02 mM sodium arsenate. The ATP inhibitors had no apparent effect upon the contraction of apical cells that had been incubated in an expanded state. In contrast, after distension of poisoned, contracted bladders, the orientation of intermediate filaments and the densities of discoidal vesicles were similar to the condition characterized by contracted cells. The results indicated that the normal reorientation of filaments, coincident with cell distension, had been suppressed by ATP inhibitors. This, in effect, impeded the filament-mediated translocation of membrane plaques to the surface. The reduction of surface area along the luminal border forced many cells to compensate by separating at their lateral margins.
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  • 152
    ISSN: 1432-0878
    Keywords: “Synaptic” ribbons ; Pineal gland ; Light exposure ; Circadian rhythm ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To characterize further the functionally enigmatic “synaptic” ribbons (SR) of the mammalian pineal gland and to study possible relationships to melatonin synthesis, in the present investigation rats were exposed to short pulses of light at night when both SR numbers and serotonin N-acetyltransferase (NAT) activity are high in comparison to day-time values. Male Sprague-Dawley rats were killed at 13:00 and 01:00 h, respectively, and at 01:10 and 02:00 h after exposure to light for 10 and 60 min, respectively. The pineals were rapidly taken out and cut sagittally in half. One half was processed for electron-microscopic quantitation of SR numbers and the other half for NAT determinations. It was found that both SR numbers and NAT activity decreased significantly when the animals were exposed to light at night. Although both parameters showed corresponding changes, there was no clear-cut correlation between SR numbers and NAT activity in individual animals within a group, except after exposure to light for 60 min when a positive correlation (R = 0.939; p 〈 0.05) existed. After exposure to light the electron-lucent vesicles of the SR decreased in number, but the length of the SR was unchanged. These results show that numbers of pineal SR can be easily and quickly manipulated and that the presently used model may be ideal in studying the poorly understood mode in which degradation of SR occurs.
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  • 153
    ISSN: 1432-0878
    Keywords: Glomerulus ; Podocytes ; Cell membrane ; Osmium tetroxide ; Dehydration ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of the podocyte membrane was studied by means of transmission electron microscopy of unosmicated tissue samples after acetone or ethanol dehydration and subsequent embedding in a polyester resin. The podocyte membrane in glutaraldehyde (GA)-fixed, acetone-dehydrated samples consisted of a relatively thick, clear layer (about 6 nm) abutted by the dark staining cytoplasm and a dark surface layer. In GA-fixed, ethanol-dehydrated samples a striking intramembranous pattern was observed in the podocyte cell membrane. The luminal podocyte membrane was regularly perforated by gaps about 25 nm wide. In grazing sections these gaps appeared round and were separated by a honeycomb pattern of intact membrane. The abluminal membrane, in contrast, generally maintained its continuity. The clear layer of the podocyte membrane was thinner in ethanol-dehydrated samples than in acetone-dehydrated ones. In tissue samples fixed with GA supplemented by ruthenium red, ethanol dehydration was not associated with cell-membrane perforations. Based on these observations as well as on biochemical data from the literature we suggest that in GA-fixed, unosmicated, acetone-dehydrated samples the structural integrity of the podocyte membrane is well preserved, while ethanol dehydration extracts some specific material from regularly distributed domains in the podocyte cell membrane.
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  • 154
    ISSN: 1432-0878
    Keywords: Vasopressin ; Oxytocin ; Opioid peptides ; Neurosecretion ; Neural lobe ; Co-localization ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The content of vasopressin, oxytocin, neurophysin, leucine-enkephalin, methionine-enkephalin, dynorphin-(1–13), and α-neoendorphin in the rat neurohypophysis was measured after different periods of dehydration and after depolarisation of isolated neural lobes and of neurosecretory nerve endings. The rates at which the amount of neurohypophysial hormone and opioid peptides decreased, and the changes in the ratios between the amount of vasopressin or oxytocin and opioid peptide in the neurohypophysis after dehydration and in the incubation medium after depolarization in vitro cast some doubt on, and can be explained by mechanisms other than co-localisation of the different peptides.
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  • 155
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    Cell & tissue research 246 (1986), S. 183-187 
    ISSN: 1432-0878
    Keywords: Neonate ; Paneth cells ; Corticosteroid ; Zinc ; Calcium ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Paneth cells containing zinc-rich granules were found in the small intestine of 6-day-old rats. These cells were more numerous in older animals and were consistently most common in the distal ileum. The zinc content of granules from 10-day-old rats was similar to that found in adults (ca 300 mg atoms/kg dry weight) but no calcium could be detected. An injection of cortisone acetate at 5 days resulted in a premature increase in the numbers of Paneth cells in 10-day-old rats. The cell granules contained normal, adult levels of zinc, a calcium concentration of ca 400 mg atoms/kg dry weight and also an increased concentration of phosphorus.
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  • 156
    ISSN: 1432-0878
    Keywords: Vasopressin ; Oxytocin ; Neurohypophysis (Median eminence, Neural lobe) ; Adrenalectomy ; Salt loading ; Salt deprivation ; Plasma volume ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In adrenalectomized rats the influence of salt loading or salt deprivation on the vasopressin and oxytocin content of the median eminence (ME) and the neural lobe (NL) was studied by means of various methods: (1) morphometric and microphotometric analysis of aldehyde fuchsin-stained sections of ME and NL; (2) immunohistochemical demonstration of neurophysin, oxytocin, and vasopressin in the ME and in the NL; (3) radioimmunological measurement of oxytocin and vasopressin in the ME and in the NL. Adrenalectomy in salt-substituted rats raised the vasopressin content of the outer layer of the ME (OLME) but had no influence on the amount of vasopressin in the inner layer of the ME and in the NL. Osmotic stimulation of adrenalectomized rats by hypertonic saline markedly diminished vasopressin and oxytocin in the inner layer of the ME and in the NL but did not, or only slightly reduced vasopressin in the OLME. Withdrawal of salt supplementation in adrenalectomized rats resulted in a decrease of plasma sodium and plasma volume. It did not change the vasopressin or oxytocin content of the inner layer of the ME and of the NL, but it was correlated with a decrease of vasopressin in the OLME. The present findings may suggest that vasopressin in the OLME is involved in salt and/or volume regulation by influencing the hypophysial-adrenal axis.
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  • 157
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    Cell & tissue research 246 (1986), S. 263-269 
    ISSN: 1432-0878
    Keywords: Spleen ; Macrophages ; Marginal metallophils ; Marginal zone ; Fluorescent analog microscopy ; Rat ; Mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The splenic macrophages of rats and mice were studied by light and fluorescence microscopy to determine their phagocytotic uptake of carbon and neutral polysaccharide (Fic-F), and their lysosomal enzyme activities. In rats, the large macrophages of the marginal zone (MZ) showed a moderate to strong acid phosphatase activity, and took up most of the Fic-F, even though they showed a weak phagocytotic activity to carbon particles. Red-pulp macrophages, however, ingested a large quantity of carbon particles, and are considered to be the major scavengers in the rat spleen. In contrast, the MZ macrophages in the mouse spleen were the major scavengers and showed a vigorous uptake of both carbon and Fic-F. In rats, the marginal metallophils (MM), located at the outer border of the periarterial lymphatic sheath and boundary between the MZ bridging channel and surrounding tissue, ingested Fic-F, whereas those located around the follicular area did not. In mice, on the other hand, the MM never ingested Fic-F. Lightly carbon-ladened small cells were constantly seen in the MZ of both rats and mice. They showed little acid phosphatase activity and did not ingest Fic-F. They were also present in the blood circulation.
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  • 158
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    Cell & tissue research 246 (1986), S. 635-640 
    ISSN: 1432-0878
    Keywords: Image analysis ; Morphometry ; Peroxisomes ; Catalase ; Cytochemistry ; Liver ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The feasibility of the application of an electronic image analyzer, the Texture Analysis System (TAS) (Leitz Wetzlar, FRG), for fast automatic ultrastructural morphometric studies of hepatic peroxisomes has been investigated. Rat liver peroxisomes were stained selectively with the alkaline DAB procedure for localization of catalase in order to obtain sufficient contrast for automatic detection by TAS. Electron micrographs of ultrathin sections from this material were analyzed both automatically by TAS and manually by using a digitizer tablet connected with an Apple IIe microcomputer. The results showed negligible differences. As far as the speed of the operation is concerned, the image analysis was 4–5 times faster than the manual technique. In further studies, the importance of using DAB-stained sections for accurate morphometric studies of peroxisomes was demonstrated by comparing the results of such DAB-stained preparations with unstained material. This revealed that the numerical density was lower and the average profile diameter higher in unstained sections. The value for volume density was also affected, being about 30% lower in such preparations. It is likely that in unstained preparations small peroxisomes without crystalline nucleoids were frequently not identified as such and were not taken into account in morphometric calculations. These observations establish that computer-controlled electronic image analysis in conjunction with selective cytochemical staining of peroxisomes for catalase provides a fast, accurate and reliable method for ultrastructural morphometric studies of this organelle in rat liver.
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  • 159
    ISSN: 1432-0878
    Keywords: Anterior pituitary gland ; Estrogen treatment ; Intercellular junctions ; Folliculo-stellate cells ; Phagocytosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural changes suggesting a phagocytic role for the nongranular folliculo-stellate cells of the anterior pituitary are investigated in estrogen-primed male rats after withdrawal of estrogen. Morphological changes in mammotropes following the removal of a subcutaneous estradiol-containing Silastic implant include the formation of intracellular lipid bodies. These lipid bodies appear to be associated with enhanced estrogen-dependent prolactin secretion in mammotropes. Seven and 24 h after estrogen withdrawal intracellular lipid within mammotropes seems to be released into the intercellular space. Seventy-two h after estrogen withdrawal, lipid droplets are almost entirely cleared from mammotropes while folliculo-stellate cells become packed with lipid globules. Folliculo-stellate cells also undergo dramatic hypertrophy 7 and 24 h after the removal of E2-containing implants. Extensive intercellular junctions including zonulae adhaerentes, desmosomes, and putative gap junctions are formed. Intercellular junctions delineate extravascular channels into which numerous microvilli project. Folliculo-stellate cells appear capable of accumulating many lipid droplets, presumably related to mammotrope metabolism. What appear to be large secondary lysosomes as well as the lipid droplets are observed within folliculostellate cells; lipid, therefore, may be degraded through a lysosomal pathway in folliculo-stellate cells.
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  • 160
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    Cell & tissue research 243 (1986), S. 91-99 
    ISSN: 1432-0878
    Keywords: Teeth ; Calcification ; Adenosine triphosphatase ; Calcium-alkaline phosphatase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Enzymatic activities of calcium-magnesium dependent adenosine triphosphatase (Ca-ATPase) and nonspecific alkaline phosphatase (ALPase) were localized at the initial calcification sites of dentin and enamel of rat incisor teeth using electron-microscopic cytochemistry. Ca-ATPase was localized in the Golgi cisternae, cytoplasmic vesicles and along the outer surface of the presecretory and secretory ameloblasts, whereas it was totally absent from the odontoblasts in the pulp. Inversely, ALPase reaction was localized along the outer surface of the odontoblasts, but almost completely absent from the ameloblasts. Diffuse extracellular reactions of both enzymes were distributed throughout the unmineralized fibrous matrix of mantle dentin in which a large number of matrix vesicles were scattered. Both Ca-ATPase and ALPase reactions, which appeared in the matrix vesicles in the process of formation of mantle dentin, became most conspicuous at the site of initial dentin calcification. At this stage, an intense Ca-ATPase reaction also appeared along some of the collagen fibrils adjacent to the reactive matrix vesicles. No ALPase reaction was localized along these Ca-ATPase reactive collagen fibrils. Our observations suggest strongly that Ca-ATPase in the matrix vesicles originates from the inner enamel epithelium and/or preameloblasts whereas ALPase originates from the odontoblasts in the pulp. The importance of the coexistence of both enzymes for the control of initial calcification of dental hard tissues is suggested.
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  • 161
    ISSN: 1432-0878
    Keywords: Calcitonin gene-related peptide ; Substance P ; Dorsal root ganglion ; Urinary bladder ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of indirect immunofluorescence, this study demonstrated the presence of calcitonin gene-related peptide-like immunoreactive (CGRPI) fibers in the bladder of the rat. These fibers were abundant in the muscle layer, in which they ran parallel to the muscles, submucosa, and epithelium. No immunoreactive cells were detected. We also examined the origins of these fibers, using a method that combined biotinized retrograde tracer (biotin-wheat germ agglutinin) (B-WGA) and immunocytochemistry. Injection of the tracer into the bladder resulted in the demonstration of small to medium-sized labeled cells that contain CGRPI structures in single dorsal root ganglion cells mostly at the level of L6 and S1, but also a few at L2. Double-staining for CGRPI and immunoreactive P-like substance (SPI) indicated that there are cells in the dorsal root ganglia at the level of L6 and S1 that react to both, but that there are many CGRPI-positive cells that contain no demonstrable SPI; most of the latter are large.
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  • 162
    ISSN: 1432-0878
    Keywords: Smooth-muscle cell ; Cell culture ; Ultrastructure ; Optical measurement ; Cell contraction ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In numerous investigations using cultured smooth-muscle cells, investigators have consistently added 10–20% fetal calf serum (FCS) to the medium to maintain viable cells. In the present study we utilized an optical technique to investigate whether smooth-muscle cells, cultured with or without FCS, maintain their contractile activity in vitro. With such optical measurement, we were able to detect signals due to spontaneous contractions, in muscle cells cultured in FCS-free medium for up to 8 days, and, for the first time, were also able to observe the conduction of these cell contractions. The ultrastructural characteristics of cultured smooth-muscle cells during contractile activity, were also examined by electron microscopy. The cells were mature and well-differentiated, and were packed with numerous myofilaments. They had developed long cell processes, and were linked to one another by gap junctions. These observations indicated that the smooth-muscle cells, cultured without FCS for 7 to 8 days, were morphologically mature and maintained their contractile activity, whereas the cells cultured in FCS-containing medium showed no detectable signs of contractile activity.
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  • 163
    ISSN: 1432-0878
    Keywords: Thyroid gland ; Corrosion casts ; Scanning electron microscopy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three-dimensional images of blood vessels in thyroid glands from normal, low iodine diet-treated, thyroid-stimulating hormone (TSH)-treated and propylthiouracil (PTU)-treated rats were investigated by use of the corrosion-cast method. The vascular casts made by the injection of methacrylate resin were observed with the scanning electron microscope. In normal animals, each follicle is surrounded by a clearly defined basket-like capillary network, which is generally independent of adjacent networks, though a few anastomoses or common capillaries are sometimes seen. In low iodine diet-treated or TSH-treated animals, the capillaries in the basket-like network become markedly dilated and fuse with one another. Though the vascular casts of PTU-treated animals are similar to those of low iodine diet-treated or TSH-treated ones in some aspects, most basket-like networks become distorted and irregular in shape, and the capillaries are heterogeneously dilated and show many buds, branches and anastomoses. We consider that these peculiar changes in the thyroid of the PTU-treated animals are due not only to the elevation of serum TSH but also to other unknown factors. It is clear that the distribution and morphology of the thyroid capillaries are extremely affected and changed by functional states of the gland.
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  • 164
    ISSN: 1432-0878
    Keywords: Calbindin-D 28K (CaBP-28K) ; Immunohistochemistry ; Growth cartilage ; Vitamin D ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of the vitamin-D dependent calcium-binding protein (Calbindin-D 28K) (CaBP-28K) in the tibial growth plate cartilage of the rat has been studied immunohistochemically using an antibody raised against rat renal CaBP-28K. The protein was detected mainly in the nuclei of chondrocytes and occasionally in the juxta-nuclear cytoplasm. The distribution was not uniform throughout the growth plate, but concentrated in the proliferatively active chondrocytes of the resting and proliferative zones. These findings raise the possibility that CaBP-28K may be involved in the mitotic activity of the chondrocytes, acting as a regulator of the proliferative process, perhaps via intranuclear calcium.
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  • 165
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    Cell & tissue research 245 (1986), S. 323-330 
    ISSN: 1432-0878
    Keywords: Rete testis ; Endocytosis ; Transferrin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The uptake and pathway of different markers and ligands for fluid-phase, adsorptive and receptor mediated endocytosis were analyzed in the epithelial cells lining the rete testis after their infusion into the lumen of these anastomotic channels. At 2 min after injection, diferric transferrin bound to colloidal gold was seen attached to the apical plasma membrane and to the membrane of endocytic coated and uncoated pits and vesicles. The injection of transferrin-gold in the presence of a 100-fold excess of unconjugated diferric transferrin revealed no binding or internalization of transferrin-gold. Similarly, apotransferrin-gold was neither bound to the apical plasma membrane nor internalized by these cells. These results thus indicate the presence of specific binding sites for diferric transferrin. At 5 min, internalized diferric transferrin-gold reached endosomes. At 15 and 30 min, the endosomes were still labeled but at these time intervals the transferrin-gold also appeared in tubular elements connected to or associated with these bodies or seen in close proximity to the apical plasma membrane. At 60 and 90 min, most of the transferrin-gold was no longer present in these organelles and was seen only exceptionally in secondary lysosomes. These results thus suggest that the tubular elements may be involved in the recycling of transferrin back to the lumen of the rete testis. The coinjection of transferrin-gold and the fluid-phase marker native ferritin revealed that both proteins were often internalized in the same endocytic pit and vesicle and shared the same endosome. However, unlike transferrin, native ferritin at the late time intervals appeared in dense multivesicular bodies and secondary lysosomes. When the adsorptive marker cationic ferritin and the fluid-phase marker albumin-gold were coinjected, again both proteins often shared the same endocytic pit and vesicle, endosome, pale and dense multivesicular body and secondary lysosomes. However, several endocytic vesicles labeled only with cationic ferritin appeared to bypass the endosomal and lysosomal compartments and to reach the lateral intercellular space and areas of the basement membrane. The rete epithelial cells, therefore, appear to be internalizing proteins and ligands by receptor-mediated and non-specific endocytosis which, after having shared the same endocytic vesicle and endosome, appear to be capable of being segregated and routed to different destinations.
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  • 166
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    Keywords: Subcommissural organ ; Ependyma ; Astrocytes ; Immunocytochemistry ; Glial fibrillary acidic (GFA) protein ; S-100 protein ; Glutamine synthetase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Antibodies raised against glial fibrillary acidic protein (GFA), S-100 protein (S100) and glutamine synthetase (GS) are currently used as glial markers. The distribution of GFA, S100 and GS in the ependyma of the rat subcommissural organ (SCO), as well as in the adjacent nonspecialized ventricular ependyma and neuropil of the periaqueductal grey matter, was studied by use of the immunocytochemical peroxidase-antiperoxidase technique. In the neuropil, GFA, S100 and GS were found in glial elements, i.e., in fibrous (GFA, S100) and protoplasmic astrocytes (S100, GS). The presence of S100 in the majority of the ventricular ependymal cells and tanycytes, and the presence of GFA in a limited number of ventricular ependymal cells and tanycytes confirm the glial nature of these cells. The absence of S100, GFA and GS from the ependymocytes of the SCO, which are considered to be modified ependymal cells, suggests either a non-astrocytic lineage of these cells or an extreme specialization of the SCO-cells as glycoprotein-synthesizing and secreting elements, a process that may have led to the disappearance of the glial markers.
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  • 167
    ISSN: 1432-0878
    Keywords: BALT ; Lymphoepithelium ; IgA response ; Ia-expression ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Changes occurring in the epithelium covering bronchus-associated lymphoid tissue (BALT) in the rat after several intratracheal administrations of horseradish peroxidase (HRP) were studied using morphological and ultrastructural methods. The epithelium is invaded by W3/ 25-positive (T-helper) lymphocytes, the BALT epithelial cells become Ia-positive and develop microvilli; there is an apparent loss of cilia. The number of non-ciliated cells in stimulated BALT increases. The non-ciliated cells can be subdivided into two cell types, one with electron-dense cytoplasm and cytoplasmic granules and the other without granules. The electron-density of the latter cell type is intermediate between that of the ciliated cells and that of the granulecontaining non-ciliated cells. The granule-containing cell types may be responsible for the uptake of antigens, while the other non-ciliated cell may be involved in the production of the secretory component and the passage of secretory IgA.
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  • 168
    ISSN: 1432-0878
    Keywords: Uropygial gland ; Sebaceous gland ; Testosterone ; Japanese quail ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of the uropygial gland of the male quail was compared to that of the sebaceous gland of the male rat after castration and testosterone treatment of both species. In intact animals, the differentiating cells of these glands displayed almost the same pattern as regards their smooth endoplasmic reticulum, an organelle involved in lipogenesis in both cases. Castration reduced the volume of this organelle, while testosterone administration restored cell morphology to a normal or supranormal level. Finally, this study showed that at ultrastructural level, there is a close functional analogy between the uropygial gland of quail and the sebaceous glands of rats as regards their androgen dependency. Consequently, the uropygial gland might be an attractive model for study of action of androgens on sebaceous-like glands.
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  • 169
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    Keywords: Organ culture ; Amelogenesis ; Dentinogenesis ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Molar tooth germs from three-day-old rats were cultured successfully for fourteen days, permitting the study of the development in vitro of both extracellular matrix and cellular elements such as odontoblasts and ameloblasts. The ultrastructure of the cultured tooth germs was compared with the ultrastructure of tooth germs in vivo at a comparable developmental stage. Progenitor cells of odontoblasts and ameloblasts were found to differentiate in vitro. Odontoblasts seemed to contain more lysosome-like bodies and fewer secretory granules than in vivo. They formed normally mineralizing dentine or a thick layer of dense, unmineralized predentine with incidentally some amorphous, extracellular material. Enamel was exclusively present opposite well developed dentine. It was often hyperor hypomineralized and enamel rods were not as regularly shaped as in vivo. In places where no enamel formation had taken place, large amounts of amorphous extracellular material were sometimes seen. From these observations it can be concluded that cellular development in cultured tooth germs appeared more or less normal, but extracellular matrix formation and mineralization were sometimes disturbed.
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  • 170
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    Cell & tissue research 246 (1986), S. 293-301 
    ISSN: 1432-0878
    Keywords: Acetylcholine ; Choline acetyltransferase (ChAT) immunocytochemistry ; Hippocampus ; Fascia dentata ; Non-pyramidal neurons ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We report here on cholinergic neurons in the rat hippocampal formation that were identified by immunocytochemistry employing a monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme. In general, ChAT-immunoreactive cells were rare, but were observed in all layers of the hippocampus proper and fascia dentata with a preponderance in zones adjacent to the hippocampal fissure and in the part of CA1 bordering the subiculum. All immunoreactive cells found were non-pyramidal neurons. They were relatively small with round or ovoid perikarya, which gave rise to thin spine-free dendrites. These hippocampal neurons were very similar to ChAT-immunoreactive cells in the neocortex of the same animals but were quite different from cholinergic neurons in the basal forebrain, medial septal nucleus, and neostriatum, which were larger and more intensely immunostained. Electron-microscopic analysis of ChAT-immunoreactive cells in the hippocampus and fascia dentata revealed synaptic contacts, mainly of the asymmetric type, on cell bodies and smooth proximal dendrites. The nuclei of the immunoreactive cells exhibited deep indentations, which are characteristic for non-pyramidal neurons. Our results provide evidence for an intrinsic source of the hippocampal cholinergic innervation in addition to the well-established septo-hippocampal cholinergic projection.
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  • 171
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    Keywords: Ontogeny ; Lymph node ; Immunohistochemistry ; Primary immune response ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The popliteal lymph nodes were removed from young rats of various ages five days after a single immunization with TNP-KLH in the hind footpads. Cryostat sections of the lymph nodes were investigated by means of enzyme and immunohistochemical techniques at the light-microscopical level. The presence and localization of anti-TNP antibody-containing cells were examined using a new technique to visualize specific antibodies. Moreover, the development of the lymph nodes following exogenous antigenic stimulation was compared with that of unstimulated lymph nodes. Specific antibody-containing cells could not be found before day 15 after birth, in rats immunized at day 10. From that time these lymphoid cells were located primarily at the border between cortex and medulla. Younger popliteal lymph nodes showed only aspecific immunoglobulin-containing lymphoid cells. With age, the number of specific antibody-containing cells tended to increase. These cells were more mature, according to morphological criteria and were located nearer the medulla. The first primary follicles were seen at day 19, as was the case in unstimulated animals. The first secondary follicles, containing germinal centers, were detected at day 23, whereas in unstimulated popliteal lymph nodes they were never found. Trapping of immune complexes could not be demonstrated before day 33 after birth. The later appearance of this phenomenon might be a consequence of the techniques applied to demonstrate specific antibody-containing cells.
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  • 172
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    Keywords: Hippocampus ; Area dentata ; Vasoactive intestinal polypeptide (VIP) ; Interneurons ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Thermic lesions in the medial septum of adult rats result in dark degeneration of terminal boutons in the stratum moleculare and hilus of the area dentata. While most of the degenerating terminals are in synaptic contact with non-reactive cells, part of them end on dendrites of VIP-like immunoreactive neurons.
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  • 173
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    Cell & tissue research 243 (1986), S. 405-414 
    ISSN: 1432-0878
    Keywords: Leydig cells ; Ethane dimethanesulphonate ; Ultrastructure ; Destruction ; Gonadotrophins ; Testosterone ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cytotoxic effects of ethane dimethanesulphonate upon rat Leydig cells were examined ultrastructurally up to 3 days after treatment and related to changes in serum levels of gonadotrophins and testosterone. Six hours after administration of ethane dimethanesulphonate the usual tubulo-vesicular morphology of Leydig-cell smooth endoplasmic reticulum was converted to small vesicles and the Golgi apparatus showed focal hypertrophy into anastomosing tubules. These changes became more marked by 12 h with many Leydig cells exhibiting karyopyknosis and hyperchromatism. Necrotic Leydig cells were often engulfed by macrophages, the latter containing pyknotic fragments of Leydig cells within their cytoplasm. One day after administration, advanced necrosis of Leydig cells occurred, many of which were phagocytosed by macrophages, and on day 3, destruction of Leydig cells was complete resulting in their elimination from the interstitial tissue, which contained only loose connective tissue and macrophages. Structural alterations to the Leydig cells from 6–24 h was reflected by a significant reduction in serum testosterone levels which further declined to the limits of detection accompanying the abolition of Leydig cells on day 3. These changes were paralleled by a significant elevation of serum LH and FSH levels suggesting diminished feedback regulation of pituitary gonadotrophin secretion. The results indicate that ethane dimethanesulphonate is a rapidly acting Leydig cell toxin which may be a useful experimental tool in further studies of spermatogenic function mediated via Sertoli cell-Leydig cell interaction.
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  • 174
    ISSN: 1432-0878
    Keywords: Skeletal muscle fibers ; Diaphragm ; Sarcolemma ; Sarcoplasmic reticulum ; Membrane contacts ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An electron-microscopic study of the myotendinous portion of the diaphragm in the Wistar rat has shown that at the ends of muscle fibers, longitudinally oriented invaginations and peripheral furrows of the sarcolemma establish specialized contacts with individual sacs of the sarcoplasmic reticulum. The construction of these terminal contacts is similar to that of contacts between sarcolemmic T-tubules and terminal cisternae of the sarcoplasmic reticulum, characterized by formation of triads. The contact zones of the sac membrane are undulated and bound to the adjoining sarcolemma via electron-dense profiles of varying forms. Frequently, the terminal contacts and triads are located at the same level within the muscle fiber, at the borderline between A- and I-bands of the sarcomeres. At the ends of muscle fibers combined contacts between peripheral furrows of the sarcolemma, terminal cisternae of the sarcoplasmic reticulum, and T-tubules of the triads are also disclosed. The implications of the terminal contacts for muscle contraction are discussed.
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  • 175
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    Cell & tissue research 243 (1986), S. 445-448 
    ISSN: 1432-0878
    Keywords: DNA turnover ; Retina ; Aging ; Photoreceptor cell ; Ganglion cell ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary DNA turnover in post-mitotic photoreceptor cells of F344 rat retina was investigated. Developing retinas of newborn rats were labelled by multiple injections of (methyl-3H)thymidine. One eye was removed on day 60 and embedded in paraffin. The groups of rats were killed 180, 365, 540 or 730 days later and the second eye was removed. Autoradiographic studies on pairs of eyes showed no detectable DNA turnover in photoreceptor cells up to the end of the experiment (near median life-span, 50% survival age). The DNA of these photoreceptor cells is not replaced through the life span of the animals; the results thus suggest that it is very stable and possibly protected in a specific manner.
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  • 176
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    Keywords: Subcommissural organ ; Glycoproteins ; Secretory process ; Immunocytochemistry ; Lectin histochemistry ; Rat ; Vertebrates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subcommissural organ (SCO) of the rat was investigated by use of histochemical and immunocytochemical methods at the light and electron-microscopic levels. Consecutive thin methacrylate sections were stained with the pseudoisocyanin (Psi), immunoperoxidase (IMC; employing an antiserum against Reissner's fiber, AFRU), periodic acid-Schiff (PAS) and periodic acid-silver methenamine (SM) techniques, and reacted with six types of lectins. Psi, SM, concanavalin A (Con A) and IMC were also used for double and triple sequential staining of the same section. Increasing dilutions of AFRU (from 1∶1000 to 1∶200 000) were used for immunostaining of serial paraffin sections. In addition, ultrastructural localization of (i) Con A-binding sites and (ii) immunoreactive secretory material was performed. Some of these procedures were also applied to the ophidian and canine SCO. Con A-positive, Psi-positive and immunoreactive materials coexisted within the same cisternae of the rough endoplasmic reticulum. The Golgi apparatus lacked Con A-positive and immunoreactive substances. Apical secretory granules and secreted material lying on the surface of the SCO showed (i) the highest affinity for AFRU, but were (ii) Con A-negative, and (iii) wheat-germ agglutinin-, PAS and SM-positive. Reissner's fiber displayed a low affinity for AFRU. It is suggested that the SCO secretes N-linked glycoproteins, the carbohydrate and protein moeities of which undergo (i) a maturation process before being released, and (ii) some kind of modification(s) after their release into the ventricle. The perivascular secretory cells of the dog SCO might secrete a material different from that secreted by the ependymal cells.
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  • 177
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    Cell & tissue research 243 (1986), S. 635-639 
    ISSN: 1432-0878
    Keywords: Mast cells ; Granule maturation ; Histamine ; Serotonin ; Heparin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Repopulation and maturation of rat mesenteric and peritoneal mast cells were studied after mast cell depletion by intraperitoneal injection of distilled water. Immature mast cells were first identified in the mesentery and peritoneal fluid 5 and 6 days, respectively, after water injection. The most immature mast cells that could be identified contained a few orthochromatic granules. Upon maturation, the granules became metachromatic and increased in size and number. Heparin, revealed by toluidine blue staining and berberine sulfate fluorescence, appeared simultaneously with orthophthaldialdehyde (OPT)-induced histamine fluorescence. Paraformaldehyde-induced serotonin fluorescence appeared somewhat later. Repopulation of mesentery and peritoneal fluid by mast cells seemed to be independent of each other and to occur from undifferentiated precursor cells.
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  • 178
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    Keywords: Hepatocytes ; Lysosomes ; Macroautophagy ; Microautophagy ; Starvation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural morphometric analysis was used to study time-dependent variations in macro and microautophagy in rat hepatocytes. Except during periods of shortterm starvation for up to 24 h, animals were kept under standardized conditions of food intake. In hepatocytes of meal-fed rats the volume fraction of macroautophagic vacuoles is significantly higher at 23:00 h, i.e., immediately before food intake, compared to 11:00 h, i.e., 12 h following feeding. During fasting, macroautophagy drops to a low level. Microautophagic vacuoles in hepatocytes of meal-fed rats, sacrificed at 11:00 or 23:00 h respectively, do not show any significant quantitative differences. However, during 12 h of starvation, the volume fraction of microautophagic vacuoles rises significantly, whereas the numerical density remains constant. Subsequently, during the second 12-h period of fasting, the volume fraction of microautophagic vacuoles remains unchanged, but the numerical density increases. Over a period of 24 h of starvation the volume fraction of the total lysosomal system does not change significantly, whereas the numerical density rises. The time-dependent changes of the macroautophagic vacuolar system correlate with the circadian, food-related variations in the protein content of individual hepatocytes from meal-fed animals. The increase in volume fraction and thereafter in number of microautophagic vacuoles, as observed during starvation, coincides with a large decrease in protein content of individual hepatocytes.
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  • 179
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    Cell & tissue research 243 (1986), S. 655-659 
    ISSN: 1432-0878
    Keywords: Nasal glands ; Protein synthesis ; Protein secretion ; Cholinergic drugs ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The secretory behaviour of rat nasal glands, under normal conditions and after the application of cholinergic drugs, has been studied using morphological and radiobiochemical techniques. Autoradiography and electrophoresis provide evidence for the selective incorporation of 3H-arginine into the glycoprotein containing fraction of the nasal glandular secretion. Radiobiochemical experiments show that labelled arginine is rapidly incorporated into the acinar cells of unstimulated glands, although it takes approximately 4 h before the labelled secretory proteins leave the cells. The secretion of proteins is stimulated by the parasympathetic agonist pilocarpine, whose main action is to promote discharge. Histological sections show a depletion of secretory granules after pilocarpine treatment. The cholinergic antagonist atropine inhibits the secretion; the acinar cells are completely filled with secretory granules following this treatment. The time course of the events following atropine administration suggests that there is no feed-back system controlling glycoprotein synthesis. The techniques employed here therefore appear to be useful for studying the effects of drugs that interfere with the secretory activity of the nasal glands.
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  • 180
    ISSN: 1432-0878
    Keywords: Histamine ; Histamine N-methyltransferase ; Immunohistochemistry ; Kidney ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Histamine N-methyltransferase (S-adenosylmethionine: histamine N-methyltransferase, E.C. 2.1.1.8) was purified to homogeneity from rat kidney, and antibody was raised against it in guinea pigs. The antibody immunoprecipitated histamine N-methyltransferase. Immunofluorescent histochemical studies with anti-histamine N-methyltransferase antibody as the first antibody and goat antiguinea pig IgG conjugated with fluorescein isothiocyanate as the second, showed the presence of immunoreactive structures in the proximal tubules of rat kidney. The brain showed no immunoreaction with the antibody.
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  • 181
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    Keywords: Catecholaminergic innervation ; TRH neurons ; Hypothalamus ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The catecholaminergic innervation of thyrotropin-releasing hormone (TRH) neurons was examined by use of a combined method of 5-hydroxydopamine (5-OHDA) uptake or autoradiography after intraventricular injection of 3H-noradrenaline (3H-NA) and immunocytochemistry for TRH in the same tissue sections at the electron-microscopic level. TRH-like immunoreactive nerve cell bodies were distributed abundantly in the parvocellular part of the paraventricular nucleus (PVN), in the suprachiasmatic preoptic nucleus and in the dorsomedial nucleus of the rat hypothalamus. In the PVN, a large number of immunonegative axon terminals were found to make synaptic contact with TRH-like immunoreactive cell bodies and fibers. In the combined autoradiography or 5-OHDA labeling with immunocytochemistry, axon terminals labeled with 3H-NA or 5-OHDA were found to form synaptic contacts with the TRH immunoreactive nerve cell bodies and fibers. These findings suggest that catecholamine-containing neurons, probably noradrenergic, may innervate TRH neurons to regulate TRH secretion via synapses with other unknown neurons in the rat PVN.
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  • 182
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    Cell & tissue research 244 (1986), S. 21-26 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Prolactin ; Tissue culture ; Fibroblasts ; Collagen ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Dispersed cells from anterior pituitary glands of male rats were cultured embedded in collagen gels or on a plastic surface. After 6 or more days in culture, cells in collagen produced more prolactin than those on plastic. The cultures in collagen had fewer fibroblasts than those on plastic. Many cells cultured in collagen developed processes, and a few cells in every culture had long extended processes that sometimes branched and resembled those of neurons. About 60% of the cells in culture including cells with well developed processes bound the monoclonal antibody A2B5, an antibody that binds to neuronal cells in culture. Fibroblasts did not have detectable binding of A2B5. Some cells with short processes reacted with antiserum to prolactin or ACTH, but the cells with neuronlike processes did not. Collagen gels may provide a matrix in which cells from the anterior pituitary gland can differentiate into neuronlike cells.
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  • 183
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    Cell & tissue research 245 (1986), S. 37-41 
    ISSN: 1432-0878
    Keywords: Astrocytes ; Dendrites ; Neuronal plasticity ; Neurosecretory neurons ; Supraoptic nuclei ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In earlier ultrastructural studies of the supraoptic nucleus in adult rats we noted “free” and incompletely covered postsynaptic densities (collectively referred to here as vacant postsynaptic densities) on dendritic shafts. Free postsynaptic densities have been reported in other parts of the central nervous system of normal rodents. We investigated the possibility that physiological activation of the supraoptic cells, which produces changes in many aspects of their morphology, would alter the incidence of the free or incompletely covered postsynaptic densities on dendrites in the supraoptic basal dendritic zone. The cells of the supraoptic nucleus are activated to increase cell firing and secretion of oxytocin and/or vasopressin in response to dehydration, gestation, and lactation. We have examined: (i) untreated virgin females; (ii) untreated males; (iii) 24 h water-deprived males; (iv) prepartum (21st day of gestation) females; (v) postpartum females (on the day of parturition); (vi) lactating females (14 days of suckling); (vii) mothers 10 days after weaning their pups; (viii) females given 2% saline to drink (dehydrated) for 10 days; and females or males given 2% saline to drink for 10 days, then given tap water for (ix) 2 or (x) 5 weeks to allow rehydration. Only long-term activation of the supraoptic nucleus by lactation or by drinking saline for 10 days brought about significant decreases in the percentage of dendrites with vacant postsynaptic densities. These densities did not reappear in saline treated rats which had been rehydrated for 2 weeks, but did return in both the 5-week rehydration and the 10-day postweaning groups. Short-term activation of the supraoptic nucleus, such as occurs at parturition or in acute dehydration, did not affect the vacant postsynaptic densities. Analysis of semiserial thin sections indicated that presynaptic elements facing the incompletely covered postsynaptic densities contain predominantly clear round vesicles and also that apparently free postsynaptic densities were usually at least partially contacted by a presynaptic ending in adjacent sections.
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  • 184
    ISSN: 1432-0878
    Keywords: Plexus choroidei ; Area postrema ; Leptomeninges ; Blood-CSF-barrier ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the borderline area between the hemal milieu of the choroid plexuses (PC) and the interstitial cerebrospinal-fluid (CSF) compartment, ground substances displaying increased amounts of basal lamina-like material and containing negatively charged sulfated glycosaminoglycans appear to be endowed with selective properties. They may function as a sieve or filtration barrier gradually controlling the passage of substances between the two milieus, depending on their charge and molecular weight. Special structural features and functional properties of ependymal cells are associated with such bordering structures. These ependymal cells are transitional elements between choroid epithelium and ciliated ependymal cells. As judged from experiments with horseradish peroxidase and conventional electron microscopy, occluding junctions at the basal pole of these cells prevent a rapid alteration in the milieu conditions, enabling gradual change from hemal to CSF composition near the bases of these transitional ependymal cells. The borderline structures between the hemal milieu of the PC and the area postrema (1) are established by leptomeningeal cells which face a hemal mileu, (2) are endowed with conspicuous tight junctions, and (3) produce a flocculent substance, the light-microscopic equivalent of which is PAS positive. These structures probably establish an effective barrier between the two milieus of different composition. The functional characteristics and the morphology of the meningeal cells facing the hemal milieu of neurohemal regions resemble closely the neurothelial cells, which are interposed between the CSF milieu and the hemal milieu in the dura mater. The present results suggest that the location between the hemal and the CSF milieu is decisive for the transformation of leptomeningeal cells into “neurothelial” elements.
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  • 185
    ISSN: 1432-0878
    Keywords: Capillary ; Epithelium ; Retina ; Retinal pigment epithelium ; Ultrastructure ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light- and urethane-induced retinopathies in rats are characterized by loss of photoreceptors. Retinal capillaries subsequently become incorporated into the normally avascular retinal pigment epithelium. These models provided an opportunity to study the response of epithelial cells to closely apposed capillaries, in order to determine if capillaries contribute to the polar organization of epithelial cells. Pigment epithelial cells reorganized their lateral plasma membrane where the latter faced intraepithelial capillaries. This normally flat, undifferentiated membrane developed attachment sites, folds and intracytoplasmic tubules, and exhibited endocytosis and putative basal lamina secretion. These structural and functional specializations are normally restricted to the basal plasma membrane — the normal vascular front of the cell facing the dense meshwork of capillaries constituting the choriocapillaris. We conclude that RPE cells, and perhaps epithelia in general, polarize in response to an adjacent capillary bed.
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  • 186
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    Cell & tissue research 245 (1986), S. 177-181 
    ISSN: 1432-0878
    Keywords: Striated muscle ; Satellite cells, muscle ; Regeneration ; Degeneration ; Culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Satellite cells were studied in the peripheral fibres from isolated rat muscles, which had been incubated for various periods of time. The cells were in an activated state with prominent organelles and increased cytoplasm visible. Mitosis of some satellite cells was occasionally observed. It is suggested that when incubated muscle preparations are used as models for physiological systems in vivo, the state of the satellite cell population should be taken into consideration.
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    Cell & tissue research 245 (1986), S. 431-437 
    ISSN: 1432-0878
    Keywords: Retina ; Arteriole ; Venule ; Tannic acid ; Peroxidase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The surface-associated vesicles in retinal arterioles and venules were studied after fixation in glutaraldehyde-tannic acid or after intravitreal injection of peroxidase or lactoperoxidase. The vesicles were concentrated along the abluminal (basal) surface of the endothelial cells and along the plasma membranes of smooth muscle cells in arterioles and of pericytes in post-capillary venules. They were rarely encountered in the deeper regions of these cells. In perpendicular sections through the cell surface the majority of vesicles were in continuity with the plasma membrane whereas in tangential sections, they appeared to lie “free” in the cytoplasm. All such vesicles were labeled after exposure to tannic acid or to the heme-proteins. Peroxidase-reaction product was never seen in the lumen of the vessels. These observations suggest that the surface vesicles in endothelial cells, smooth muscle cells and pericytes are invaginations of the plasma membrane and are thus not involved in the transcytosis or endocytosis of proteins. The vesicles in the latter two cell types may be involved in some aspect of contractility rather than pinocytosis.
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  • 188
    ISSN: 1432-0878
    Keywords: VIP ; Nerve fibers ; Ganglion cells ; Chromaffin cells ; Ultrastructure ; Adrenal gland ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary VIP-like immunoreactivity was revealed in a few chromaffin cells, medullary ganglion cells and a plexus of varicose nerve fibers in the superficial cortex and single varicose fibers in the juxtamedullary cortex and the medulla of the rat adrenal gland. VIP-like immunoreactive chromaffin cells were polygonal in shape without any distinct cytoplasmic processes and they appeared solitarily. Their cytoplasm contained abundant granular vesicles having a round core and the immunoreactive material was localized to the granular core. VIP-immunoreactive ganglion cells were multipolar and had large intracytoplasmic vacuoles. The immunoreactive material was localized not only in a few granular vesicles but also diffusely throughout the axoplasm. VIP-immunoreactive varicose nerve fibers in the superficial cortex were characterized by abundant small clear vesicles and some large granular vesicles, while those in the juxtamedullary cortex and medulla and the ganglionic processes were characterized by abundant large clear vesicles, as well as the same vesicular elements as contained in the nerves in the superficial cortex. The immunoreactive material was localized on the granular cores and diffusely in the axoplasm in both nerves. Based on the similarity and difference in the composition of the vesicles contained in individual nerves, it is likely that the VIP-immunoreactive nerve fibers in the medulla and the juxtamedullary cortex are derived from the medullary VIP-ganglion cells, while those in the superficial cortex are of extrinsic origin. The immunoreactive nerve fibers in both the cortex and the medulla were often in direct contact with cortical cells and chromaffin cells, where no membrane specializations were formed. The immunoreactive nerve fibers were sometimes associated with the smooth muscle cells and pericytes of small blood vessels in the superficial cortex. In addition they were often seen in close apposition to the fenestrated endothelial cells in the cortex and the medulla, only a common basal lamina intervening. Several possible mechanisms by which VIP may exert its effect in the adrenal gland are discussed.
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  • 189
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    Cell & tissue research 245 (1986), S. 591-598 
    ISSN: 1432-0878
    Keywords: Taxol ; Microtubules ; Mitosis ; Meiosis ; Testis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The effect of taxol, an inhibitor of microtubule degradation, on the seminiferous epithelium was studied. Taxol arrested spermatogenesis at metaphase in both mitotic and meiotic germ cell division. Microtubules were seen to accumulate, especially in the cytoplasm of the spermatogonia, and also in the early spermatids and Sertoli cells. No microtubule accumulation was observed in germ cells during meiotic prophase. Formation of the flagellum was affected in developing spermatids. Peculiar lamellar structures, probably derived from degenerating mitochondria, were seen in the cytoplasm of late spermatids and Sertoli cells. The results are compared with the effects of other mitotic inhibitors such as colchicine and vinca alcaloids.
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  • 190
    ISSN: 1432-0878
    Keywords: Sensory neuron ; Glycoprotein ; Glycoconjugate ; Lectin cytochemistry ; Myelin ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Paraffin sections of the trigeminal nerve root of the rat, and human spinal nerve root and trigeminal ganglion were stained with a battery of lectin-horseradish peroxidase conjugates to localize and characterize glycoconjugate (GC) in situ. In the rat the myelin sheath of the peripheral segment contained GC with sialic acid most probably linked to the penultinate disaccharide galactose(β1 → 4)-N-acetylglucosamine (Gal(β1 →)-GlcNAc), and complex type N-glycosidic side chains. The myelin sheath in the central segment differed in containing little if any of the GC named above and in containing GC with terminal β-Gal linked to N-acetylgalactosamine (GalNAc), terminal GalNAc and fucose. Schwann cells stained for GC with GlcNAc or mannose whereas oligodendroglia stained for GC with the terminal disaccharide Gal-(β1 → 3)-GalNAc and N-glycosidic side chains, especially in presumed Golgi zones, but also in processes continued as the outer myelin sheath. The human myelin sheath in the central segment differed from that of the rat in not staining with lectins specific for fucose and terminal GalNAc. Sialic acid and terminal β-Gal were seen in the human central segment but these sugars appeared to bind to astroglial structures rather than to the myelin sheath as in the rat. Astrocytes in both rat and man were stained by two fucose-binding lectins. Several lectins revealed affinity for GC in the neurilemmal sheath, and staining of this structure was stronger in the human specimens. Neurons in the human trigeminal ganglion ranged from unstained to strongly positive for fucoconjugate in cytoplasmic bodies and plasmalemma. Positive ganglion cells gave rise to unmyelinated fibers which also stained for fucoconjugate. Remak fibers and their extensions into the substantia gelatinosa of the human spinal cord stained strongly for content of fucose. The stronger lectin affinity for N-glycosidic core sugars in the peripheral as compared with the central segment suggests that lectins localize Po protein in peripheral myelin. The reactivity for several sugars in the central segment can possibly be attributed to myelin-associated glycoprotein (MAG) of central myelin, but lectin staining for GalNAc shows in addition a biochemically unrecognized GC with O-glycosidic linked oligosaccharides in myelin. The lectin cytochemistry indicates that the 170 K Dalton glycoprotein with PNA affinity obtained from rat sciatic nerves occurs in nodes of Ranvier.
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  • 191
    ISSN: 1432-0878
    Keywords: Catechol-O-methyltransferase ; Macrophage ; Ovary ; Corpus luteum ; Oviduct ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Catechol-O-methyltransferase (COMT) (EC 2.1.1.6) was localized in rat ovary, oviduct, and uterus using immunocytochemical methods. Immunoreactive deposits were found in the cytoplasm of macrophages in the ovary, epithelial cells of the oviduct, and glandular epithelial cells of the non-pregnant uterus. The pattern of localization observed in the extraneuronal elements suggests that enzyme may function in extraneuronal inactivation of catechols in the ovary, oviduct, and uterus.
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  • 192
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    Cell & tissue research 245 (1986), S. 635-641 
    ISSN: 1432-0878
    Keywords: Hepatocytes ; Bile canaliculi ; Contractility ; Phalloidin ; Cytochalasin B ; Actomyosin ; Tissue culture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In short-term cultures of rat hepatocytes, bile canaliculi enclosed between unseparated cell couplets are able to perform periodical contractions resulting in expulsion of bile. Pericanalicular cytoskeletal proteins are involved in canalicular contractility: F-actin, myosin and tropomyosin are associated around bile canaliculi, as revealed by staining with tetramethylrhodaminyl-phalloidin and by immunofluorescence. Bile canalicular contractility is distributed by cholestatic agents that are known to interfere with actin polymerization; e.g., phalloidin and also cytochalasin B inhibit canalicular contractility and cause pericanalicular vacuolization and formation of blebs. Whereas the association of the cytoskeletal proteins is not affected by treatment with cytochalasin B, treatment with phalloidin results in dissociation of F-actin and myosin, indicating that binding of phalloidin to F-actin impairs its molecular interaction with myosin.
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  • 193
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    Cell & tissue research 245 (1986), S. 649-655 
    ISSN: 1432-0878
    Keywords: Ethane dimethanesulphonate ; Leydig cells ; Destruction ; Unilateral cryptorchidism ; Regeneration ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Leydig cells in testes of adult rats were selectively destroyed by a single intraperitoneal injection of ethane dimethane sulphonate. Four days later rats were made unilaterally cryptorchid and 1, 2 and 4 weeks later the histology of the testes was examined by light microscopy and morphometry. After induction of unilateral cryptorchidism, the volume of abdominal compared to scrotal testes was reduced by 45–60% due to rapid impairment of spermatogenesis in abdominal testes. Leydig cells were not present in either scrotal or abdominal testes in the 1-week unilateral crytorchid group. A new generation of foetal-type Leydig cells was observed in scrotal testes of the 2-week unilateral crytorchid group although their total volume per testis estimated by morphometry, was small, being approximately 1 μl. In contrast, the abdominal testis exhibited a remarkable proliferation of foetal-type Leydig cells (total volume per testis, 16 μl) which predominantly surrounded the peritubular tissues of the seminiferous tubules. A similar morphology and pattern of Leydig cell development was observed in scrotal and abdominal testes of the 4-week unilateral cryptorchid group where total Leydig cell volume was 7 μl vs 21 μl, respectively. The results show that regeneration of a new population of Leydig cells occurs more rapidly in the abdominal testis than in the scrotal testis of the same animal. These observations suggest the possibility that augmentation of Leydig cell growth is mediated by local intratesticular stimulatory factors within the abdominal testis. Development of new Leydig cells from the peritubular tissue provides circumstantial evidence that the seminiferous tubules and in particular the Sertoli cells, are a likely source of agents that stimulate the growth of Leydig cells.
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  • 194
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    Cell & tissue research 246 (1986), S. 523-530 
    ISSN: 1432-0878
    Keywords: Enamel organ ; Papillary cell ; Annular gap junction ; Lysosome ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary To investigate the mechanisms whereby annular gap junctions in the papillary cells of the enamel organ are degraded intracellularly, continuously growing rat incisors were examined by electron microscopy of routine thin sections as well as for the cytochemical localization of inorganic trimetaphosphatase activity. Routine thin-section analysis revealed small flat or undulated gap junctions, hemi-annular gap junctions between an invaginated cell process and a cell body, and fully internalized cytoplasmic annular gap junctions. Both hemi-annular and annular gap junctions usually contain various organelles and/or inclusions, such as mitochondria, endoplasmic reticulum, ribosomes, vesicles, and lysosomes in the cytoplasm confined by the junctional membranes. Annular gap junctions are sometimes fused with vesicular or tubulovesicular structures. Cytochemistry of inorganic trimetaphosphatase activity revealed an intense enzymatic reaction within a system of tubular structures and round or oval dense bodies. Both structures are believed to correspond to primary lysosomes. A part of the Golgi apparatus also shows a weak reaction. Although hemi-annular gap junctions never show enzymatic reaction, annular gap junctions sometimes contain reaction products throughout their interior cytoplasm and inclusions. Fusion of annular gap-junctional membranes with reaction-positive tubular structures is also observed. In one instance, revealed in serial sections, an annular gap junction was encircled entirely by a reaction-positive structure. These results suggest that cytoplasmic annular gap junctions are formed by endocytosis of hemi-annular gap junctional membranes from the cell surface and then degraded intracellularly by lysosomal enzymes.
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  • 195
    ISSN: 1432-0878
    Keywords: Endocytosis ; Absorptive cells ; Kidney (proximal tubule cells) ; Apical tubules ; Membrane recycling ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using horseradish peroxidase (HRP) as a tracer, we have investigated if the so-called apical tubules (AT) in the kidney proximal tubule cells are directly involved in the endocytic process by carrying the tracer into the cells, or if they are derived from the intracellular membrane compartments. Rat kidney was fixed by vascular perfusion at different time intervals after intravenous injection of HRP and prepared for electron microscopy. An analysis revealed that 0.5 min after injection, invaginations of the plasma membrane and small apical endocytic vesicles, including coated vesicles, were labelled with reaction product, whereas almost all large apical endocytic vacuoles and the AT were negative. The endocytic vacuoles and about 18% of the AT were labelled 1 min after injection. The reaction product in the large endocytic vacuoles was usually seen along the luminal surface of the vacuoles. The AT with reaction product appeared as a branched network, and were frequently connected with the labelled endocytic vacuoles. Three min after injection, reaction product was detected in about 38% of the AT, and thereafter, the percentage increased to about 74% after 7 min. No reaction product was detected in the Golgi complex at any time after HRP-injection. These findings indicate that the AT are probably formed by budding off from the large endocytic vacuoles, rather than being directly involved in the endocytic process.
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  • 196
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    Cell & tissue research 243 (1986), S. 145-155 
    ISSN: 1432-0878
    Keywords: Endothelium ; Lectin-receptors ; Choriocapillaris ; Eye ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The choriocapillaris is one example of a capillary bed lined by a fenestrated endothelium that is restrictive to exogenous tracers and endogenous plasma proteins. In this study we have examined the distribution of cell-surface monosaccharides utilizing biotinylated lectin-avidin ferritin cytochemistry. Receptors for wheat germ agglutinin were localized to the plasmalemma and diaphragms of some fenestrae, vesicles, and channels at the luminal endothelial front in amounts greater than seen for the other lectins employed. The absence of labeling following inhibition with N-acetylglucosamine and after tissue digestion with N-acetylhexosaminidase, but not after neuraminidase indicated that this lectin marked N-acetylglucosamine residues and not sialic acid. Wheat germ agglutinin receptors were not affected by pronase E or trypsin digestion, but were partially removed by proteinase K. The latter also removed many fenestral diaphragms. Wheat germ agglutinin receptors were cleaved with endoglycosidase D. The combined results indicate that the wheat germ agglutinin receptor is of the low-mannose type and part of a protein with hydrophobic properties. Receptors for concanavalin A (mannose) and Ricinus communis agglutinin (galactose) were also localized to the plasmalemma and endothelial diaphragms. The examination of sections at different tilt angles revealed that these lectins bound to the endothelium in a non-random distribution, encircling diaphragms of fenestrae and channels. Soybean agglutinin (N-acetylgalactosamine) marked endothelial structures sparsely. Following digestion with pronase E or trypsin, receptor sugars for the latter three lectins were completely removed, indicating their presence on protease susceptible glycoproteins. These findings demonstrate that the endothelium of the choriocapillaris bears carbohydrate moieties that are different than those described for permeable fenestrated endothelia.
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  • 197
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    Cell & tissue research 243 (1986), S. 157-164 
    ISSN: 1432-0878
    Keywords: Endothelium ; Choriocapillaris ; Heparin ; Heparan sulfate ; Cytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location and chemical composition of anionic sites on the endothelium of the choriocapillaris was investigated with cationic ferritin and enzyme digestion techniques. Cationic ferritin administered intravenously initially labeled essentially all fenestral diaphragms. Within 30 min after injection, no diaphrams remained labeled, but they could be relabeled by a second cationic ferritin injection. Following perfusion of cationic ferritin, the entire luminal front of the endothelium was labeled: the plasmalemma and fenestral, vesicle, and channel diaphragms. Perfusion of neuraminidase or chondroitinase did not affect subsequent cationic ferritin binding. In contrast, heparitinase removed anionic sites on all structures except fenestral diaphragms. Cationic ferritin did not mark the endothelium following heparinase digestion. All sites were cleaved with pronase E. These results indicate that heparin is the anionic moiety on fenestral diaphragms while the glycocalcyces of the plasmalemma and vesicle and channel diaphragms are rich in a heparan sulfate proteoglycan. Furthermore, since the heparan sulfate localized to these structures was digested by both heparinase and heparitinase, it is in a form similar to heparin. These findings demonstrate that the endothelium of the choriocapillaris bears cell-surface anionic components that are different than those described for fenestrated endothelia lining other vascular beds.
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  • 198
    ISSN: 1432-0878
    Keywords: Hepatocytes ; Liver ; Protein mass ; Starvation ; Proteolysis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary With regard to the protein content, as analysed cytophotometrically, of hepatocytes from rats kept under a 12L 12D photoperiod (photophase 7:00–19:00), the following facts have been established: 1) Hepatocytes of different classes of ploidy all demonstrate, more or less equally, daily variations in protein content and also its reduction after 24-h fasting. 2) With computer analysis of data obtained at eight time points during a period of 24 h, a sinusoidal curve of the protein content of individual mononuclear tetraploid hepatocytes throughout the day could be demonstrated with a maximum at 6:20 and a minimum at 18:20. 3) Animals, fed with meals via a dispensing machine from 23:00 to 24:00 only, show a similar sinusoidal curve but with higher amplitude, and a virtually identical mean value as those fed ad libitum. The maximum was found at 10:40, revealing a time lag of 12 h after food intake, the minimum at 22:40. 4) Trained animals deprived of food during the standardized feeding time revealed a moderate reduction of their hepatocyte protein content in the first 6 h, then a 6-h period with a steep fall followed by a slower reduction. After 24 h, the mean hepatocyte protein mass had decreased to 72% of that at the commencement of fasting at 23:00.
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  • 199
    ISSN: 1432-0878
    Keywords: Azygos vein ; Cardiac muscle ; Smooth muscle ; Cell culture ; Capillarization ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The azygos vein of the rat can be divided into three regions: 1) The proximal cardiac region, where the wall is composed of two and sometimes three layers of cardiac muscle and a thin discontinuous layer of smooth muscle cells. Vascular casts of this region demonstrate layers of capillaries closely following the orientation of the cardiac fibres. 2) A transitional zone, where both cardiac and smooth muscle cells interdigitate. In this zone, close associations between smooth muscle and cardiac muscle cells can be observed, however, gap junctions do not appear to be present. 3) Beyond this transitional zone the vessel resembles a typical thin-walled vein. The cells of the media of the entire length of azygous vein have been isolated and grown in culture and two separate viable populations identified corresponding to smooth and cardiac muscle.
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  • 200
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    Cell & tissue research 243 (1986), S. 229-238 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Corticotropes ; α-MSH ; Immunocytochemistry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific antisera to α-melanotropin (α-MSH) and corticotropin (ACTH 1-39) were used to obtain immunocytochemical evidence for the differential localization of α-MSH and ACTH in the secretory granules of corticotropes of rat anterior pituitary. The specificity of the antisera was established by binding 131I-labeled α-MSH and ACTH 1-39 to their respective antisera. Double-labeling immunocytochemistry (for α-MSH, ferritin; for ACTH, colloidal gold) was performed. Some secretory granules were labeled with ferritin particles (α-MSH), whereas others contained gold particles (ACTH). Only a few granules showed both ACTH and α-MSH. In typical corticotropes (stellate in form with a small number of secretory granules aligned along the cell periphery) only some of the secretory granules that were labeled with anti-ACTH serum were also immunoreactive to anti-α-MSH. In atypical corticotropes (polygonal in shape and containing a large number of secretory granules) almost all of the immunoreactive ACTH secretory granules were also positive to anti-α-MSH serum. An intermediate type of corticotrope was observed containing a small number of secretory granules, almost all of which were labeled with anti-α-MSH. Thus, rat anterior pituitary corticotropes may be classified into three types according to the distribution and content of α-MSH. The light-microscopic immuncytochemistry provided similar results.
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