Gene/Protein Disease Symptom Drug Enzyme Compound
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Microridges produce a characteristic fingerprint-like pattern on the surface of fish oral mucosa. The cytoskeleton in these microridges was examined by immunofluorescence microscopy and transmission electron microscopy after detergent extraction and decoration with myosin subfragment 1. The effect of cytochalasin B on microridges was probed with scanning electron microscopy. Immunofluorescence microscopy revealed that actin filaments were present throughout the periphery of the epithelial cells and were especially localized beneath the free surface of the epithelium. In thin sections treated with Triton X-100, the majority of filaments in the microridges and their bases were found to be actin filaments and a plexus of keratin filaments that underlay the network of actin filaments. A part of the plexus of keratin filaments entered the microridges. After extraction with Triton X-100 and decoration with myosin subfragment 1, decorated actin filaments were found in the microridge cores, connected to the keratin filaments. The keratin filaments aggregated in the pattern of microridges and a few of them protruded into the microridges. Treatment with cytochalasin B caused microridges to disappear or to become thinner and lower or to change short or microvillus-like microridges. When most microridges disappeared, the surface of the superficial cells was prominently swollen, but the cell boundaries were fastened, and the microridges in the periphery were preserved. On the basis of these observations, the possible roles of actin and keratin filaments in the maintenance and the formation of microridges are discussed.
Anat Rec 1991 Jun
PMID:Cytoskeleton in microridges of the oral mucosal epithelium in the carp, Cyprinus carpio. 171 56

A murine monoclonal IgM antibody, M3, which interferes with both polymorphonuclear leukocyte (PMN) phagocytosis and bactericidal activity, was used to examine the subcellular location of antigens bearing 3-fucosyllactosamine (CD15 antigens) within this cell type. Percoll gradient-separated secondary granule fractions were rich in CD15 antigens, with at least seven antigens recognizable in SDS-PAGE/electroblot studies. Sonication/sedimentation experiments using secondary granule fractions showed that both soluble and sedimentable CD15 antigens were present. Exposure of purified PMN to the secondary granule secretagogue phorbol myristate acetate caused extracellular release of two or three CD15 antigens, which could be purified by immunoprecipitation using antibody M3. Triton X-114 phase-partition experiments showed that secondary granule fraction CD15 antigens could be partitioned into hydrophilic (aqueous phase) and hydrophobic (detergent phase) antigens, suggesting that several of these antigens were integral secondary granule membrane components. Ultrastructurally, PMN intracellular granules showed two patterns of CD15 expression, localization over both granule matrix/granule membrane and localization to only granule membrane. Colocalization studies showed that lactoferrin and CD15 antigens were both present in a subset of intracellular granules, confirming a secondary granule location for these antigens.
Anat Rec 1990 Nov
PMID:Functional, physical, and ultrastructural localization of CD15 antigens to the human polymorphonuclear leukocyte secondary granule. 197 22

A procedure has been developed for the three-dimensional immunoelectron microscopic localization of cytoskeletal filaments by a deep-etching replica method in combination with immunogold labeling and/or myosin subfragment 1 (S1) decoration techniques. Neonatal hamster heart cells grown on glass coverslips were extracted with Triton X-100 or physically permeabilized by breaking open the cell membranes. S1 decoration was performed on some specimens immediately after the permeabilization. After prefixation in formaldehyde, samples were immunostained with poly- or monoclonal antibodies to desmin or vimentin, and indirectly tagged with colloidal gold probes by the biotin-streptavidin method. After postfixation with glutaraldehyde, tannic acid and osmium tetroxide, the cells were freeze-etched and rotary-replicated with platinum and carbon in a freeze-fracture apparatus. Replicas were viewed with a transmission electron microscope using a tilting specimen stage to obtain stereo images. The procedure made it possible to identify the specific filaments within the complex cytoskeletal networks in cultured hamster heart muscle and nonmuscle cells at high resolution and in three dimensions. The method has advantages in its three-dimensionality and feasibility to evaluate the data by comparing them with those obtained by alternative light microscopic methods. Details of the protocol and a description of the results of using three different antibodies are given.
Anat Rec 1991 Mar
PMID:Deep-etching immunogold replica electron microscopy of cytoskeletal elements in cultured hamster heart cells. 202 81

Actin filaments, intermediate filaments, and microtubules in the odontoblasts of rat incisors were investigated electron microscopically using heavy meromyosin and taxol. Actin filaments were abundant at the periphery of the odontoblast process in the form of a network or in bundles. In a branch of the odontoblast process, longitudinally oriented actin filament bundles were found. Most actin filaments were associated with the plasma membrane via electron-dense material which stained with tannic acid. The intermediate filaments had a diameter of 11 to 13 nm. They were distributed throughout the cytoplasm of odontoblasts. They ran lengthwise in the core of the odontoblast process, which showed a different distribution compared with that of actin filaments. Microtubules, which were disrupted after Triton X-100 but preserved by addition of taxol, tended to be associated with intermediate filaments. Such a relation was also seen in conventional preparations. Coated vesicles, which were abundant at the periphery of the odontoblast process, were often associated with actin filaments. Therefore, it is suggested that actin filaments, in the odontoblast process at least, play a role associated with the coated vesicles at the periphery of the process, and may be involved in coated vesicle transport.
Anat Rec 1987 Oct
PMID:Microtubules, intermediate filaments, and actin filaments in the odontoblast of rat incisor. 368 67

The lungs of male LAF1 mice were locally irradiated with doses of 5, 9, and 13 Gy. The animals were killed at times corresponding to the appearance of histologically identifiable fibrosis or, for 13 Gy, at the LD50 for these doses and strain of mouse: 63, 36, and 28 weeks postirradiation (PI) respectively. Lungs were excised, incubated in buffer alone, or partially digested with enzymes for determination of relative glycosidase resistance, fixed with ruthenium red/Triton X-100 for demonstration of basal laminar anionic sites, and processed for electron microscopy. Sham-irradiated and untreated control groups (0 Gy, 0 times) were also processed. Tissue was examined ultrastructurally and alterations in both alveolar and capillary basal laminar anionic sites were quantitated. In each of the doses examined the number of anionic sites surpassed normal levels; however, the glycosidase resistance of the regenerated laminae at these late time points was not significantly altered from controls. This contrasts with the marked increase in the glycosidase resistance of laminae regenerating from radiation damage (4-12 weeks PI) reported earlier. The increased numbers of anionic sites were compared to expected values derived from models based on compensatory synthesis and continued accumulation and indicate close correlation with certain aspects of the compensatory synthesis model but not with others. The effects on basal laminar permeability, basal laminar thickening, and fibrotic induction are discussed.
Anat Rec 1986 Jun
PMID:Cell-cell matrix interactions in induced lung injury: III. Long term effects of X-irradiation on basal laminar proteoglycans. 372 10

Sciatic nerves from young mice were incubated for 2-8 hours in 0.5% Triton X-100 in 0.5 M ammonium acetate, a solution which solubilizes the large and small basic proteins of the myelin sheath. As previously noted (Peterson and Gruener, 1978), myelin sheaths from treated nerves extensively split and unravelled along major dense lines. Small focal areas of compact myelin remained. In freeze-fracture replicas, areas of myelin with lamellar splitting contained few intramembranous particles, while membrane areas with greater than normal densities of particles were associated with the patches of compact myelin membrane. Fixation for as short a time as 15 minutes stabilized the myelin membrane enough to prevent the Triton X-100 effects, even when incubations were extended to 20 hours. Controls, both untreated and 0.5 M ammonium acetate-treated nerves, had predominantly compact myelin sheaths; their leaflets were covered with numerous intramembranous particles. The data suggest that Triton X-100 alters the compact structure of peripheral nervous system myelin. In areas where lamellae are split and separated, there is a loss of intramembranous particles. It appears that the loss of intramembranous particles is related to the removal of the basic proteins which are located in major dense line regions of compact myelin sheaths.
Anat Rec 1983 Dec
PMID:Electron microscopic study of intramembranous changes in protein-extracted peripheral nervous system myelin. 667 Jul 54