Sawada H, Stukenbrok H, Kerjaschki D, Farquhar M G
Am J Pathol. 1986 Nov;125(2):309-18.
The distribution of podocalyxin was determined by several different immunoelectron microscopic methods, which included so-called diffusion methods carried out on cryostat sections and surface methods done on ultrathin sections. When localization was carried out by three different indirect immunogold methods (using ultrathin frozen sections, ultrathin sections cut from tissues embedded in Lowicryl K4M or LR White, or 20- mu cryostat sections for incubations), gold particles were found exclusively on the urinary surfaces of glomerular epithelial cells and, at a lower concentration, on the luminal surface of glomerular endothelial cells. No gold was bound to the basal surfaces (soles) of the epithelial foot processes, the filtration slit diaphragms, the glomerular basement membrane (GBM), or those regions of the endothelial plasmalemma that face the GBM. When podocalyxin was localized by an indirect immunoperoxidase method on cryostat sections, heavy deposits of reaction product were seen on the same cell surfaces as with the immunogold methods. Each of the methods used had certain advantages as well as limitations, but the collective results obtained were convergent and complimentary. It is concluded that podocalyxin is restricted in its distribution to the urinary surfaces of epithelial cells and the luminal surface of endothelial cells; it is missing or present in very low concentrations on the soles (basal surface) of the epithelial foot processes, the slit diaphragms and the basement membrane surface of the endothelium; and the podocalyxin-containing regions of the epithelial and endothelial plasmalemmae constitute microdomains of distinctive membrane protein composition on the corresponding cell surfaces.
采用几种不同的免疫电子显微镜方法确定了足萼糖蛋白的分布,这些方法包括在冷冻切片上进行的所谓扩散法以及在超薄切片上进行的表面法。当通过三种不同的间接免疫金法(使用超薄冷冻切片、从包埋在Lowicryl K4M或LR White中的组织切取的超薄切片或20微米的冷冻切片进行孵育)进行定位时,金颗粒仅在肾小球上皮细胞的尿极表面被发现,在较低浓度下也出现在肾小球内皮细胞的管腔表面。在上皮足突的基底表面(底部)、滤过裂隙膜、肾小球基底膜(GBM)或内皮细胞质膜面对GBM的那些区域均未发现金结合。当通过间接免疫过氧化物酶法在冷冻切片上对足萼糖蛋白进行定位时,在与免疫金法相同的细胞表面观察到大量反应产物沉积。所使用的每种方法都有一定的优点和局限性,但获得的总体结果是一致且互补的。结论是,足萼糖蛋白的分布局限于上皮细胞的尿极表面和内皮细胞的管腔表面;在上皮足突的底部(基底表面)、裂隙膜和内皮细胞的基底膜表面缺失或含量极低;上皮和内皮细胞质膜中含有足萼糖蛋白的区域在相应细胞表面构成了具有独特膜蛋白组成的微区。