Fiskum G, Craig S W, Decker G L, Lehninger A L
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3430-4. doi: 10.1073/pnas.77.6.3430.
Treatment of isolated rat hepatocptes with low concentrations of digitonin increases the permeability of the plsma membrane to cytosolic proteins without causing release of organelles such as mitochondria into the surrounding medium. Electron microscopy showed that treatment of the cells with increasing concentations of digitonin results in a progressive loss in the continuity of the plasma membrane, while most other aspects of cellular morphology remain normal. Depletion of background staining material from the cytosol by digitonin treatment of the cells greatly enhances the visualization of the cytoskeleton. The use of this technique, together with immunofluorescent light microscopy, has verified the presence of an actin-containing filamentous network at the hepatocyte cortex as well as intermediate filaments distributed throughout the cell. Digitonin is thus useful both for selectively permeabilizing the plasma membrane and for intensifying the appearance of intracellular structures such as microfilaments that are normally difficult to observe in cells such as hepatocytes.
用低浓度的洋地黄皂苷处理分离的大鼠肝细胞,可增加质膜对胞质蛋白的通透性,而不会导致线粒体等细胞器释放到周围培养基中。电子显微镜显示,用浓度不断增加的洋地黄皂苷处理细胞会导致质膜的连续性逐渐丧失,而细胞形态的大多数其他方面仍保持正常。通过用洋地黄皂苷处理细胞来耗尽胞质中的背景染色物质,可大大增强细胞骨架的可视化效果。这项技术与免疫荧光光学显微镜一起使用,已证实肝细胞皮质存在含肌动蛋白的丝状网络以及分布于整个细胞的中间丝。因此,洋地黄皂苷不仅可用于选择性地使质膜通透,还可用于增强细胞内结构(如微丝)的外观,而这些结构在肝细胞等细胞中通常难以观察到。