Nadezhdina E S, Vaĭsberg E A
Tsitologiia. 1987 May;29(5):543-8.
Cultured pig kidney epithelial cells were centrifuged at 20,000 gav so that the centrifugation force was oriented parallel to the substrate, fixed and processed for indirect immunofluorescent staining with tubulin and vimentin antibodies. After a 2 hour centrifugation vimentin filaments aggregated in the centripetal parts of the cells (probably, because of their association with floating lipid vesicles). Microtubule-organizing centers were found near the centripetal poles of the nuclei, which migrated in the direction of the centrifugal force. The distribution of the cytoplasmic microtubules did not change during centrifugation. The staining of the cultures one hour after centrifugation revealed vimentin-containing spots with radiating intermediate filaments in most of the cells. These spots were localized near the cell nuclei; double immunofluorescent staining with tubulin and vimentin antibodies showed that their position was identical to that of the microtubule-organizing centers. Similar foci of vimentin filaments were seen in the cells after a 3-4 hour centrifugation. Probably, these structures participate in organizing the intermediate filament cytoskeleton in cells.
将培养的猪肾上皮细胞以20,000 gav离心,使离心力与底物平行,固定并用微管蛋白和波形蛋白抗体进行间接免疫荧光染色处理。离心2小时后,波形蛋白丝聚集在细胞的向心部分(可能是因为它们与漂浮的脂质囊泡相关联)。在细胞核的向心极附近发现了微管组织中心,其朝着离心力方向迁移。离心过程中细胞质微管的分布没有变化。离心1小时后对培养物进行染色,发现大多数细胞中存在含有波形蛋白的斑点,并有放射状的中间丝。这些斑点位于细胞核附近;用微管蛋白和波形蛋白抗体进行的双重免疫荧光染色表明,它们的位置与微管组织中心相同。离心3 - 4小时后,在细胞中也观察到了类似的波形蛋白丝聚集点。这些结构可能参与细胞中间丝细胞骨架的组织形成。