Koury S T, Repasky E A, Eckert B S
Cell Tissue Res. 1987 Jul;249(1):69-77. doi: 10.1007/BF00215420.
Cytoskeletons of primitive erythrocytes have been isolated from the embryos of day 12 pregnant C57/Bl mice and examined by transmission electron microscopy, immunofluorescence microscopy, and SDS-polyacrylamide gel electrophoresis. Microtubules are the most prominent cytoskeletal component. They are found either singly or organized into loose bundles just under the plasma membrane, but do not form classical marginal bands in most cells. Immunofluorescence with a polyclonal tubulin antiserum confirms this distribution and further reveals numerous mitotic figures among cells. Rhodamine-conjugated phalloidin and heavy meromyosin labeling reveal that actin is localized in the cortex of the primitive erythrocyte in the form of 6 nm filaments. Antibody directed against avian erythrocyte alpha spectrin demonstrates that spectrin is also found in the cortex. Occasional 10-nm intermediate filaments, observed in the primitive erythrocytes by electron microscopy, are believed to be of the vimentin class based on positive reaction of the cells with vimentin-specific antiserum. In addition, a band in erythrocyte cytoskeletons comigrates in SDS-polyacrylamide gels with vimentin isolated from mouse kidney. Spectrin and actin were also found to be associated with the membrane of primitive erythrocytes when membrane ghost preparations were analyzed by SDS-polyacrylamide gel electrophoresis.
已从怀孕12天的C57/Bl小鼠胚胎中分离出原始红细胞的细胞骨架，并通过透射电子显微镜、免疫荧光显微镜和SDS-聚丙烯酰胺凝胶电泳进行了检查。微管是最突出的细胞骨架成分。它们要么单独存在，要么在质膜下方组织成松散的束状，但在大多数细胞中不形成经典的边缘带。用多克隆微管蛋白抗血清进行免疫荧光证实了这种分布，并进一步揭示了细胞中有许多有丝分裂图像。罗丹明偶联的鬼笔环肽和重酶解肌球蛋白标记显示，肌动蛋白以6纳米细丝的形式定位于原始红细胞的皮质中。针对禽红细胞α-血影蛋白的抗体表明，血影蛋白也存在于皮质中。通过电子显微镜在原始红细胞中偶尔观察到的10纳米中间丝，基于细胞与波形蛋白特异性抗血清的阳性反应，被认为属于波形蛋白类别。此外，红细胞细胞骨架中的一条带在SDS-聚丙烯酰胺凝胶中与从小鼠肾脏分离的波形蛋白一起迁移。当通过SDS-聚丙烯酰胺凝胶电泳分析膜空壳制剂时，还发现血影蛋白和肌动蛋白与原始红细胞膜相关。
