Madri J A, Roll F J, Furthmayr H, Foidart J M
J Cell Biol. 1980 Aug;86(2):682-7. doi: 10.1083/jcb.86.2.682.
Affinity-purified rabbit antibodies specific for two large noncollagenous gycoproteins--laminin and fibronectin--were used to study the distribution of these proteins in normal murine kidneys. Immunofluorescence staining of conventional frozen sections demonstrates fibronectin within mesangial areas of the glomerulus. Laminin is also found in mesangial areas. However, it also appears to be distributed in typical basement membranelike patterns on glomerular and tubular basement membranes and Bowman's capsule. At the ultrastructural level, by labeling 600-800-A thick frozen sections with a three-stage procedure consisting of specific antibodies, biotinyl sheep anti-rabbit IgG, and avidin-ferritin conjugates, fibronectin is present ony in the mesangial matrix and is specifically localized to areas immediately surrounding mesangial cell processes. Laminin, on the other hand, is found uniformly distributed throughout tubular basement membranes, the mesangial matrix, and Bowman's capsule. In glomerular basement membranes, laminin labeling is restricted to the lamina rara interna and adjacent regions of the lamina densa.
用针对两种大型非胶原蛋白糖蛋白(层粘连蛋白和纤连蛋白)的亲和纯化兔抗体来研究这些蛋白质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,纤连蛋白存在于肾小球的系膜区。层粘连蛋白也存在于系膜区。然而,它似乎也以典型的基底膜样模式分布在肾小球和肾小管基底膜以及鲍曼囊上。在超微结构水平上,通过用由特异性抗体、生物素化羊抗兔IgG和抗生物素蛋白-铁蛋白缀合物组成的三步法标记600-800埃厚的冷冻切片,发现纤连蛋白仅存在于系膜基质中,并特异性定位于紧邻系膜细胞突起的区域。另一方面,层粘连蛋白均匀分布于整个肾小管基底膜、系膜基质和鲍曼囊中。在肾小球基底膜中,层粘连蛋白标记仅限于内疏松层和致密层的相邻区域。