Roll F J, Madri J A, Albert J, Furthmayr H
J Cell Biol. 1980 Jun;85(3):597-616. doi: 10.1083/jcb.85.3.597.
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.
用针对I型、III型、AB2型胶原蛋白以及一种来自小鼠肿瘤的部分特性已明确的IV型胶原蛋白的亲和纯化兔抗体,来研究胶原蛋白在正常小鼠肾脏中的分布。对常规冰冻切片进行免疫荧光染色显示,I型和III型胶原蛋白存在于大血管周围的束状结构以及肾小球和肾小管周围的纤维中,而IV型和AB2型胶原蛋白则以线性方式分布在肾小管、肾小球和鲍曼囊的基底膜以及系膜柄中。通过使用特异性胶原蛋白抗体、生物素化羊抗兔IgG和抗生物素蛋白-铁蛋白缀合物的三步法,对600至800埃厚的冰冻切片进行染色,在超微结构水平上检查IV型和AB2型胶原蛋白的分布。通过该方法进行的标记显示,AB2型和假定的IV型胶原蛋白在所有三种基底膜中均共分布。此外,系膜基质显示含有这两种胶原蛋白类型。这些结果支持了基底膜中胶原蛋白异质性的最新生化证据,也支持了系膜基质与肾小球基底膜之间存在结构关系的概念。