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Codistribution of collagen types IV and AB2 in basement membranes and mesangium of the kidney. an immunoferritin study of ultrathin frozen sections

机译:IV型和AB2型胶原蛋白在肾脏的基底膜和肾小球膜中的共分布。超薄冰冻切片的免疫铁蛋白研究

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摘要

Affinity-purified rabbit antibodies specific for collagen types I, III, AB2 and for a partially characterized type IV collagen derived from a murine tumor were used to study the distribution of collagens in the normal mouse kidney. Immunofluorescence staining of conventional frozen sections demonstrated that types I and III were present in bundles around large vessels and in fibers surrounding glomeruli and tubules, whereas types IV and AB2 were distributed in a linear fashion along basement membranes of tubules, glomeruli, and Bowman's capsule and in the mesangial stalk. The distribution of types IV nd AB2 was examined at the ultrastructural level by staining of 600- to 800-A thick frozen sections with a three-stage procedure employing specific collagen antibodies, biotinyl sheep antirabbit IgG, and avidin-ferritin conjugates. Labeling by this procedure demonstrated codistribution of types AB2 and the putative type IV in all three basement membranes. In addition, mesangial matrix was shown to contain both of these collagen types. These results support recent biochemical evidence of collagen heterogeneity in basement membranes, and also support the concept of a structural relationship between mesangial matrix and glomerular basement membranes.
机译:使用亲和纯化的特异性针对I,III,AB2型胶原蛋白和源自鼠类肿瘤的部分表征的IV型胶原蛋白的兔抗体来研究胶原蛋白在正常小鼠肾脏中的分布。常规冷冻切片的免疫荧光染色显示,I型和III型存在于大血管周围的束中以及肾小球和肾小管周围的纤维中,而IV型和AB2型则沿着肾小管,肾小球和鲍曼氏囊的基底膜呈线性分布。在肾小球系膜上通过使用特定胶原蛋白抗体,生物素羊抗兔IgG和抗生物素蛋白-铁蛋白结合物的三阶段程序,对600-800-A厚的冷冻切片进行染色,在超微结构水平上检查了IV nd AB2型的分布。通过该程序标记证明在所有三个基底膜中AB2型和推定的IV型共同分布。此外,显示肾小球系膜基质同时包含这两种胶原类型。这些结果支持最新的生化证据表明基膜中胶原异质性,也支持肾小球系膜基质与肾小球基膜之间结构关系的概念。

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