Zhou Y, Moszczynska A, Opas M
Department of Anatomy and Cell Biology, Faculty of Medicine, University of Toronto, Canada.
Exp Eye Res. 1994 May;58(5):585-93. doi: 10.1006/exer.1994.1053.
We have generated antisera to pig retinal pigment epithelial (RPE) cells and to their fractionated membrane proteins. The antisera have been screened functionally by adhesion inhibition assays, and morphologically with immunofluorescence microscopy of cultured cells and frozen sections of the retina. From these sera, we have affinity purified five monospecific antibodies to the RPE surface molecules having molecular weights of 220, 180, 110, 85 and 70 kDa, which effectively inhibit cell-substratum attachment. The 220 and 85 kDa antigens are localized to focal contacts of the cultured cells and are seen in patches in the intact RPE layer in frozen sections of eye tissues. Immunofluorescence microscopy shows that the 180, 110 and 70 kDa antigens localize predominantly to cell-cell junctions. Hence, these antigens may participate in both cell-cell and cell-substratum adhesion in RPE cells. Staining patterns obtained with confocal microscopy of frozen sections of the retina demonstrate that these antibodies are RPE-specific. These adhesion-related molecules may play a role in maintenance of the morphological and functional integrity of RPE.
我们制备了针对猪视网膜色素上皮（RPE）细胞及其分离的膜蛋白的抗血清。通过黏附抑制试验对这些抗血清进行了功能筛选，并利用培养细胞的免疫荧光显微镜检查和视网膜冰冻切片进行了形态学分析。从这些血清中，我们亲和纯化了五种针对RPE表面分子的单特异性抗体，这些分子的分子量分别为220、180、110、85和70 kDa，它们能有效抑制细胞与基质的黏附。220 kDa和85 kDa的抗原定位于培养细胞的黏着斑，在眼组织冰冻切片的完整RPE层中呈斑块状可见。免疫荧光显微镜检查显示，180、110和70 kDa的抗原主要定位于细胞间连接。因此，这些抗原可能参与RPE细胞中的细胞间黏附和细胞与基质的黏附。用视网膜冰冻切片的共聚焦显微镜检查获得的染色模式表明这些抗体具有RPE特异性。这些与黏附相关的分子可能在维持RPE的形态和功能完整性方面发挥作用。
