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Rat anterior pituitaries were cytologically studied following cultivation in organ culture, with and without the addition of hypothalamic and cortical extracts. Although five distinct cell types could be identified with classical stains in the uncultivated glands, the peroxidase-labeled antibody technique (using antibodies against STH, LTH, FSH, LH and TSH) showed that not all of the immune-specific cell types were being identified with the classical stains. This discrepancy was magnified following culture as chromophilic cells seen with classic stains decreased in number with an increase in culture time. The peroxidase technique, however, revealed that all cells remained constant in type and number regardless of time in culture. While the addition of either hypothalamic or cortical extract to the culture medium produced cytological alterations demonstrated by the classical dyes, the antibody technique showed no such alterations. Such a comparison of staining techniques emphasizes the hazards of relying solely on histological procedures to reveal the hormonal activity of the pituitary gland.
Anat Rec 1975 Jan
PMID:Immunochemical staining of the rat adenohypophysis in organ culture. 4 77

The morphology and development of junctional complexes between blastomeres of the preimplantation rabbit embryo were investigated using several approaches. Electron microscopic examination of embryos stained en bloc with uranyl acetate, and the study of junction permeability using horseradish peroxidase and lanthanum nitrate provided information on structure, intermembrane spacing and permeability of the junctional complexes. In addition, the freeze fracture technique was used with day 5 and day 6 blastocysts, since the large size of these embryos facilitated use of this method. These experiments showed that although rudimentary junctions were present between blastomeres of the early cleavage stages, effective tight junctions were not present until the blastocyst stage. Electron microscopic examination of thin sections revealed apical foci of membrane approximation or "fusion" between trophoblast cells by day 4. Freeze fracturing revealed a lattice of interconnecting ridges (on the A face) and grooves (on the B face) in the apical region between trophoblast cells of the day 5 blastocyst. This lattice formed a continuous band along the apical margin of each cell, and therefore constituted a zonula occludens. The zonula occludens of the day 5 blastocyst averages 2-3 ridges per lattice, while day 6 blastocysts had lattices that averaged 5-6 ridges. Also seen in the freeze fracture replicas from the day 5 and day 6 blastocysts were local accumulations of intramembranous particles on the A face. These particles were often observed in aggregates similar to those of previously described gap junctions. It could not be determined whether these small regions of particles were true gap junctions or a possible primitive form of gap junction because the complementary pitted surfaces (B face pits) were not demonstrated.
Anat Rec 1975 Jan
PMID:Junctional complexes in the preimplantation rabbit embryo. 4 78

The yolk sac placenta has been implicated previously in transmission of passive immunity to the fetus. This work uses an immunohistochemical technique devised by Sternberger et al. ('69) to study this problem. Rabbit serum containing gamma globulins was injected into the uterine lumen of the White Swiss mouse during the last third of pregnancy. Two or four hours later yolk sacs were removed and fixed in 2% paraformaldehyde or freeze-dried and fixed with paraformaldehyde vapors. Finely chopped tissue was treated with (1) sheep antiserum to rabbit serum gamma globulin, (2) an antigen-antibody complex consisting of horseradish peroxidase and rabbit anti-horseradish peroxidase and (3) hydrogen peroxide and 3,3'-diaminobenzidine. Reaction product was heavily concentrated in visceral yolk sac endodermal cells, frequently deposited in endothelial cells of vitelline vessels, and rarely in the serosal basement membrane and mesothelial cells which border the exocoelomic cavity. This supports evidence of other workers that the yolk sac membrane rapidly absorbs substances with which it comes in contact, transport into vitelline vasculature is a route of transfer from mother to fetus, and possible transfer into the exocoelomic cavity and thence to the amniotic cavity may occur in mice.
Anat Rec 1976 Apr
PMID:Localization of rabbit gamma globulins in the mouse visceral yolk sac placenta. 5 9

The extracellular space of mouse ovarian follicles and stroma contained horseradish peroxidase (HRP) reaction product one minute after intravenous injection of the tracer. In addition to pinocytotic uptake of the tracer, non-vesicular staining with HRP reaction product, heretofore unrecognized, was noted in a variety of ovarian cell-types. This diffuse staining was correlated with changes in cellular morphology suggestive of degeneration. These findings are discussed in relation to the composition of cell populations comprising follicles and ovarian stroma and alterations in cell function.
Anat Rec 1979 Mar
PMID:Differences in mouse ovarian cells as distinguished by horseradish peroxidase labelling. 8 20

The basement membranes in the exocrine pancreas were examined by routine electron microscopy, by fixation for electron microscopy with a glutaraldehyde-tannic acid solution and by staining for ultrastructural demonstration of mucosubstances with the dialyzed iron, high iron diamine, ruthenium red and Concanavalin A-horseradish peroxidase procedures. The basement membranes are considered from morphologic and histochemical observations to consist of an inner lamina lucida, an intermediate lamina densa and an outer lamina diffusa. Sulfated mucosubstance was found in the lamina diffusa of the basement membrane of all epithelial cells and capillary endothelial cells but was encountered in the lamina lucida of the duct cells exclusively. Bridging structures, presumably polypeptides, were also seen connecting the lamina densa and the basal plasma membrane in specimens fixed with the glutaraldehydetannic acid solution. The findings demonstrated that the histochemical and morphological qualities of the basement membrane are uniform of a given cell type but differ considerably among the several cell types in the pancreas.
Anat Rec 1977 Jul
PMID:The ultrastructural histochemistry of the basement membranes of the exocrine pancreas. 14 23

Nerves in the tongues of adult and immature rats were examined with respect to their permeability to exogenous cytochrome-c (mol wt 12,000) injected into the tongue. The distribution of cytochrome-c was determined in cryostat sections on the basis of the peroxidase activity of this protein. Nerves of 14-day-old rats were permeable to injected cytochrome-c. The larger nerves of older animals showed only localized accumulations of cytochrome-c reaction product both between and within axons adjacent to endoneurial blood vessels. Reaction product was not found, however, in association with blood vessels penetrating nerves of the tongue that were not within the limits of tracer spread. In the smallest nerve brances, thin linear strands of reaction product filled the interstices between the nerve fibers.
Anat Rec 1975 Apr
PMID:Permeability barriers to cytochrome-c in nerves of adult and immature rats. 16 39

Tight junctions (zonulae occludentes) create a pericellular barrier to the diffusion of large molecules in non-keratinizing mammalian epithelia. However, in cornifying epithelia such as the epidermis, the importance of tight-junctional elements versus secreted intercellular lipid for barrier function is uncertain. In an attempt to resolve this question, we compared membrane structure in the stratum granulosum and stratum corneum of epidermis, esophagus, and vagina of newborn and adult humans and mice under both normal and various experimental conditions. We incubated pieces of epidermis in organ culture and infused tissues with lanthanum or horseradish peroxidase in vivo and in vitro. All were processed for electron microscopy of freeze-fracture replicas or thin sections. Lanthanum seeped outward to the stratum granulosum in all tissues examined--further apical migration was halted by lamellar-body contents in skin. A similar pattern of intercellular lamellar lipid deposition and membrane structure occurred in all epithelia studied. Freeze-fracture replicas of these obstructive regions revealed occasional, incomplete junctional strands (particularly in moist epithelia) and abundant lamellar material, but complete zonulae occludentes were never encountered. A possible relationship between moisture and tight junction formation was further suggested by organ culture experiments during which brief incubations stimulated an increase in the number of junctional strands and diminished numbers of lamellar bodies. We conclude that, in the epithelia studied, the deposition of secreted lamellar body contents forms the barrier to water-soluble tracer loss: tight-junctional elements are either absent or too fragmentary to constitute an effective barrier.
Anat Rec 1977 Dec
PMID:Membrane alterations during cornification of mammalian squamous epithelia: a freeze-fracture, tracer, and thin-section study. 33 80

3H-concanavalin A and the concanavalin A-horseradish peroxidase staining technique were used to study the distribution of surface coat material on the epithelium of the nasal folds and nasal groove of mouse embryos. In stages shortly before and during epithelial fusion concanavalan A stained or labeled material was present at apical surfaces of epithelial cells of the nasal groove and nasal folds. Silver grains, representing bound 3H-concanavalin A, were counted in defined areas of the nasal groove and presumptive fusion area in both anterior and posterior regions of the nasal folds. For both stages examined there was a significant increase in the amount of 3H-concanavalin A bound by presumptive fusion areas in posterior regions of the nasal folds as compared with anterior regions; i.e., the atact between the nasal folds. This finding is consistent with results from investigations of palatal shelf and neural fold fusion which suggest that increased synthesis of surface coat material is associated with adhesion and fusion of epithelial folds and shelves.
Anat Rec 1979 Feb
PMID:Distribution of surface coat material on nasal folds of mouse embryos as demonstrated by concanavalin A binding. 42 93

Horseradish peroxidase introduced into the lumen of the rat epididymis was taken up by the columnar cells of the epithelium by five minutes and more so after longer periods. The apical cells and particularly the clear cells in the caput and cauda epididymis, respectively, showed significantly greater endocytotic activity than the principal cell in both locations. Within 14 days after castration, however, such differences in absorptive activity among the various cell types were essentially obscured because of increased endocytosis by the androgen-deficient principal cells. The results are discussed briefly in terms of the function of different epithelial cell types and secretory/absorptive activity in the epididymis.
Anat Rec 1979 Feb
PMID:The differential absorptive activity of epithelial cells of the rat epididymus before and after castration. 42 1

Perisinusoidal (P.S.) cells occurring in the spaces of Disse in the livers of normal cats, dogs, miniature pigs, albino rats, human adults and children were examined by electron microscopy. The ultrastructural details of the P.S. cells and their topographic relationships with hepatocytes, sinusoidal lining cells and reticulum fibers are described. Species differences between P.S. cells were primarily a dissimilarity in lipid content: the main ultrastructural features were the same in all species studied. The P.S. cells of the rat liver displayed only low endocytotic activity, and no phagosome formation following intravenous administration of horseradish peroxidase. The close topographic relationship of the P.S. cells with the intralobular reticulum fibers was reminiscent of the intimate connection between fibroblasts and collagen fibers, or, in cat liver, of the reticulum cell--reticulum fiber association seen in lymphoid organs. Fibroblasts were not found inside the hepatic lobules. These findings support the conclusion that the reticulum fibers of hepatic lobules are produced by perisinusoidal cells which, however, display also other functions.
Anat Rec 1979 Aug
PMID:Ultrastructure of the hepatic perisinusoidal cells in man and other mammalian species. 47 17


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