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The distribution and density of receptors for concanavalin A (Con A) on the surfaces of cells of intact and isolated popliteal and axillary lymph nodes were investigated in the rabbit. Intact lymph nodes were perfused via the subcapsular (marginal) sinus with either Con A peroxidase or Con A ferritin, fixed with glutaraldehyde, and processed for electron microscopy. Both Con A peroxidase and Con A ferritin were distributed on the plasmalemma of lymphocytes, macrophages, neutrophils, plasma cells, reticular endothelial cells, and the vascular endothelium. Counts of Con A-conjugated ferritin particles indicated that the density of Con A receptors was generally similar for lymphocytes, macrophages, and neutrophils but lower on plasma cells. When lymph node cells were isolated by mechanical methods and exposed to Con A ferritin, the label was homogenously distributed on the cell surfaces of most cells. However, Con A binding was significantly higher on the surface of isolated cells than in the intact node. It is suggested that the increase in density of Con A binding sites on isolated cells may possibly be due to an unmasking of cell surface moieties in which additional Con A receptor sites become available as a result of the isolation procedure. The density of Con A ferritin binding sites was also significantly lower on the surface of isolated plasma cells than the lymphocyte and macrophage, suggesting that the density distribution of cell surface saccharides is different for various lymphoid cells.
Anat Rec 1982 Sep
PMID:Concanavalin A receptor sites on lymph node cells in vivo and in vitro. 629 43

The distribution of lysozyme in the endocervix of estrous, pseudopregnant, and ovariectomized rabbits was studied using two different immunocytochemical techniques--the unlabeled antibody enzyme method of Sternberger et al. (1970) and the peroxidase-labeled antibody method of Taylor and Burns (1974). With both procedures, a fine immunostaining precipitate was seen over the entire area of basal mucous granules, while immunodeposits were coarser and mostly located in the outer zone of central and apical granules. A nonspecific staining was noted when tissues were reacted with peroxidase-antiperoxidase complex alone. This troublesome artifact was abolished by preincubating tissues with human IgA. This step did not affect the specific immunostaining for lysozyme yet nonspecific staining was absent from specificity and method controls carried out for both immunocytochemical procedures. The presence of high levels of lysozyme in the endocervical epithelium of estrous rabbits was also confirmed in enzymatically isolated endocervical epithelia using the lysoplate method of Osserman and Lawlor (1966). Mucous granules and immunostainable intracellular lysozyme were abundant during estrus, decreased during early pseudopregnancy, and were absent after long-term ovariectomy. However, they were restored by the administration of estradiol (5 micrograms/12 hours/10 days) to ovariectomized animals. These data indicate a common hormonal regulation and secretory mechanism for endocervical mucous glycoproteins and lysozyme.
Anat Rec 1984 Aug
PMID:Ultrastructural immunocytochemical localization of lysozyme in the mucociliary epithelium of the rabbit endocervix in different hormonal states. 638 22

The endocytic activity of the low cuboidal cells lining the rete testis was analyzed by electron microscopy following injection of various tracers into the lumen of these anastomotic channels. At 1 and 5 minutes after injection, cationic ferritin (CF) and concanavalin A-ferritin (Con A) were seen bound to the apical plasma membrane and to the membrane of subjacent vesicles or invaginations connected to this apical membrane. At 30 and 60 minutes, these tracers were found in intracytoplasmic vesicles and in vesicles connected to the lateral or basal plasma membrane as well as in the lateral intercellular space and in the lamina lucida of basal lamina. At 30 minutes, CF and Con A also appeared in the matrix of pale multivesicular bodies while at 1 hour dense multivesicular bodies were labeled. At 2 hours and later time intervals, the tracers accumulated in dense granules identified as lysosomes. Native ferritin (NF), concanavalin A-ferritin in presence of alpha-methyl-D-mannoside, and horseradish peroxidase or albumin bound to colloidal gold were all to be incorporated by the lysosomal system of these epithelial cells, as just described for CF and Con A, but these various tracers were not bound to the apical plasma membrane or to the membrane of cytoplasmic vesicles, nor were they found in the intercellular spaces or the lamina lucida at the base of the cells. Thus, the epithelial cells of the rete testis do not appear to be only involved in the uptake of substances from the lumen and their disposal by the lysosomal system, but also appear to contribute to the transport of certain macromolecules from the lumen to the laterobasal surfaces of the cells. These cells may thus play a role in determining the composition of the rete testis fluid.
Anat Rec 1984 Jun
PMID:Endocytosis in epithelial cells lining the rete testis of the rat. 646 29

In order to study the distribution pattern of specific antibody-containing cells in the spleen of rabbits during the secondary immune response, rabbits were given two intravenous injections of either free or liposome-associated human serum albumin (HSA) within an interval of 2 months. Demonstration of specific antibody-containing cells was performed by incubation of sections of spleen with HSA-horseradish peroxidase (HRP) conjugates, followed by peroxidase cytochemistry. Specific anti-HSA antibody-containing cells were detected already within 2 days after booster and peak numbers were found 4 days after booster. The bulk of these cells localized in the coaxial lymphocyte sheaths surrounding the terminal arterioles in the spleen. Specific antibody-containing cells were also found in the follicles. Using a double immunoenzyme technique we demonstrated that a majority of the specific antibody-containing cells produced immunoglobulin G(IgG) antibodies. From the results, it is also concluded that, after a priming injection with liposome-associated HSA, liposomes do not further enhance the secondary immune response, when they are also used for the booster injection.
Anat Rec 1984 Apr
PMID:The development of specific antibody-containing cells in the spleen of rabbits during the secondary immune response against free or liposome-associated albumin antigen. 661 Mar 67

Through the combined use of peroxidase cytochemistry and examination at the ultrastructural level, the present study has identified liver hemopoietic foci containing three forms of erythropoietic cells, two forms of myelopoietic cells, and a population of peroxidase nonreactive cells within the extravascular compartments of mouse fetal liver. The nonreactive cells were 10 micron in diameter, displayed no peroxidase activity and were designated type I cells. This cell had an irregular nucleus, small profiles of rough endoplasmic reticulum (RER), a considerable population of monoribosomes and a few polyribosomes. The incidence of this cell type decreased significantly from 50% at 12 days gestation to approximately 10% of the hemopoietic cells at 17 days gestation. Type I cells could not be classified into a hemopoietic lineage and may represent undifferentiated hemopoietic stem cells. Three forms of erythropoietic cells, designated types II, III, and IV, were identified. These cells had a diffuse cytoplasmic peroxidase reaction, no peroxidase positive membrane-bound organelles, and were approximately 7 micron in diameter. They corresponded to the more classically defined proerythroblast, polychromatophilic erythroblast, and nucleated normoblast, respectively. Types II and III had moderate cytoplasmic reactions, whereas type III, in addition, had a slight nuclear reaction. Type IV cells had a very dense cytoplasmic reaction but no nuclear reaction. Of the myelopoietic cells detected, one form had a slightly reactive Golgi and a few reactive granules. The other form possessed a clearly positive nuclear envelope (NE), RER, Golgi, and a population of reactive granules. The phagocytic sinusoidal lining cells (Kupffer cells) were peroxidase negative in contrast to similar cells in the rat. A population of peroxidase-positive granules was detected in fetal liver developing hepatocytes at 17 days gestation and increased in number with age. The morphology and organization of these various cell types in the liver hemopoietic environment are discussed.
Anat Rec 1983 Sep
PMID:The liver hemopoietic environment: II. Peroxidase reactive mouse fetal liver hemopoietic cells. 663 32

Thyrotropin-releasing hormone (TRH) stimulates prolactin production in cultured GH3 rat anterior pituitary tumor cells. For correlation of cell-by-cell prolactin distribution and intracellular hormone concentration, GH3 cells were grown to plateau-phase density on glass coverslips in plastic dishes. Acetone-fixed, cell-bearing coverslips were stained for prolactin by an immunoglobulin-peroxidase bridge technique (Mason et al., '69); cells on the plastic dishes were assayed for prolactin (microcomplement fixation immunoassay, Tashjian, '73) and protein content. Intracellular prolactin, unaffected quantitatively by acetone fixation and choice of substratum, was localized immunocytochemically by a granular brown precipitate, abolished if anti-prolactin serum was preabsorbed with rat prolactin or omitted from the protocol. Intracellular prolactin was maximized with colchicine (5.0 X 10(-6) M; final 3 hr of incubation) in control and TRH-treated (10 ng/ml; 48 hr) GH3 cell cultures. A total of 8,500 cells were classified by light microscopy as unstained, heavily (H) or moderately (M) stained for prolactin. In controls, 35% of cells were prolactin-positive: 5% H and 29% M. After TRH, 45% were positive: 7% H and 38% M. Although prolactin-positive cells were unevenly distributed, comprising 25% to 46% of cells in individual microscopic fields in controls, TRH increased the proportion of M cells in all areas. TRH treatment raised prolactin levels to 450% of control, but mathematical analysis attributed less than 30% of the increase to new prolactin-positive cells. We conclude that TRH acts on GH3 cultures principally by raising the mean hormone content of individual positive cells rather than by increasing the proportion of cells committed to prolactin production.
Anat Rec 1980 Jun
PMID:Immunocytochemical analysis of prolactin production by monolayer cultures of GH3 rat anterior pituitary tumor cells: I. Long-term effects of stimulation with thyrotropin-releasing hormone (TRH). 677 30

Carbonic anhydrase was localized in osteoclasts by light and electron microscopy using a preembedding peroxidase-antiperoxidase staining method. Osteoclasts on the endosteal surface of the metatarsi of calcitonin-treated and untreated (control) chicks were studied. A positive staining reaction was seen in the cytosol, in the Golgi apparatus, in and on vesicles, on the plasma membrane of the ruffled border, and on the bone surface beneath control osteoclasts. After calcitonin treatment, positive staining reactions were seen at the same sites except that staining was absent on the plasma membrane and endosteal bone surface. The morphology of osteoclasts from calcitonin-treated chicks was indicative of cell inactivity. The carbonic anhydrase which was bound to the plasma membrane of the ruffled border is appropriately arrayed for hydrogen ion secretion and subsequent mineral dissolution. The presence of the enzyme within lysosomelike vesicles and on the endosteal surface beneath active cells suggests that it may be released into the resorbing zone along with the lysosomal hydrolases. The function of the extracellular enzyme is also unknown.
Anat Rec 1982 Sep
PMID:Ultrastructural immunocytochemical localization of carbonic anhydrase in normal and calcitonin-treated chick osteoclasts. 681 93

Porphyrin- and endogenous peroxidase-containing accumulations of cytoplasmic inclusions in astrocytes of the hypothalamic periventricular and arcuate nuclei and periventricular areas of the lateral ventricles except for ependymal astrocytes were observed by fluorescent microscopy and histochemical techniques in the wild type, C57BL/6J, and tabby mice. These cells can be also visualized with phase contrast and dark-field techniques or by staining sections embedded in polymerized resin with toluidine blue. The brains of heterozygous females and hemizygous males for the blotchy allele (mottled locus on the X-chromosome) failed to show this specific class astrocytes. The findings described are possibly due to defective copper metabolism in mottled mutants which may include a number of other related abnormalities, including reduced activity of copper-dependent enzymes such as porphyrin-containing peroxidases. Sexual difference in the number of the accumulations in the anatomically and physiologically normal tabby mouse was clearly expressed.
Anat Rec 1983 Mar
PMID:Structural and cytochemical changes in astrocytes from the brain periventricular zone of the copper-deficient blotchy mouse. 683 46

Fluorescence spectroscopy has been employed to investigate the emission spectrum of unusual orange-red fluorescence found in the mouse optic nerve. Comparison of the spectra obtained with those of a number of porphyrins used as standards (protoporphyrin, uroporphyrin, and coproporphyrins) shows that the autofluorescence excited at about 400 nm (Soret band) is due to the presence of a mixture of these or other porphyrins in the nerve. Phase contrast, dark-field, and light-microscopy techniques demonstrated that the fluorescence is emitted by dense, coarse inclusions in the cytoplasm of astrocytes. The inclusions also exhibit high activity of endogenous peroxidase, a heme (porphyrin)- containing enzyme, characteristic for process of phagocytosis. A possible participation of these astrocytes in phagocytosis is delineated.
Anat Rec 1983 Mar
PMID:Light-microscopic and microspectrofluorometric characterization of porphyrin-containing astrocytes in mouse optic nerve. 683 47

This study has examined the fine structure and some cytochemical characteristics of the endodermal and mesothelial cells of the rhesus monkey yolk sac between 25 and 66 days of gestation. The endodermal cells were characterized by abundant granular endoplasmic reticulum, some agranular endoplasmic reticulum, a well-developed Golgi apparatus, and numerous large mitochondria. During the earlier part of the period studied, endodermal cells had a few acid phosphatase and arylsulfatase-positive lysosomes and moderate numbers of catalase-positive microperoxisomes. During the later stages of development, large granules (believed to be lysosomes) with a heterogeneous content were numerous in the cytoplasm. Mesothelial cells showed fewer development changes. Throughout this period they were usually flattened cells with long microvilli, small mitochondria, and limited amounts of granular endoplasmic reticulum. The mesothelial cells had acid phosphatase reaction product in the Golgi region and occasional large vesicles, but were negative for arylsulfatase and catalase. One specimen was incubated at 37 degrees C in the presence of horseradish peroxidase in order to examine endocytosis. Both the mesothelial cells and endodermal cells internalized the peroxidase into a variety of cytoplasmic vesicles. Based on their cytology, the endodermal cells may function in the synthesis of serum proteins during this period, as has been suggested in other species. They may also be involved in lipid metabolism. The mesothelial cells appeared less synthetically active, but evidence suggested that they may be involved in collagen and extracellular matrix production. The endocytic activity displayed by both cell types may indicate a role in fluid and metabolite transfer across the epithelia. The cytology of both cell types was very similar to that described for human yolk sacs, suggesting that the rhesus monkey may be a useful species in which to study the maturation of yolk sac function.
Anat Rec 1983 Feb
PMID:A fine structural and cytochemical study of the rhesus monkey yolk sac: endoderm and mesothelium. 684 66


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