THE CYTOSKELETON OF ISOLATED MURINE PRIMITIVE ERYTHROCYTES
THE CYTOSKELETON OF ISOLATED MURINE PRIMITIVE ERYTHROCYTES
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DOI：
10.1007/bf00215420
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发表时间：
1987-01-01
影响因子：
3.6
通讯作者：
ECKERT, BS
中科院分区：
文献类型：
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作者：
KOURY, ST;REPASKY, EA;ECKERT, BS
从妊娠第12天的C57/Bl小鼠胚胎中分离出原始红细胞的细胞骨架，并通过透射电镜、免疫荧光显微镜和sds -聚丙烯酰胺凝胶电泳进行检测。微管是最重要的细胞骨架成分。它们或单独存在，或组织成松散的束，就在质膜下，但在大多数细胞中不形成经典的边缘带。多克隆微管蛋白抗血清的免疫荧光证实了这种分布，并进一步揭示了细胞中许多有丝分裂的图形。罗丹明偶联的phalloidin和大量的meromyosin标记显示肌动蛋白以6 nm细丝的形式定位于原始红细胞的皮层。针对禽红细胞α spectrin的抗体表明，在皮层中也发现了spectrin。电镜观察到原始红细胞中偶有10 nm的中间细丝，根据细胞与vimentin特异性抗血清的阳性反应，认为是vimentin类。此外，红细胞骨架中的一个条带在sds -聚丙烯酰胺凝胶中与从小鼠肾脏分离的波形蛋白相结合。用sds -聚丙烯酰胺凝胶电泳法对鬼影膜进行分析，发现鬼影蛋白和肌动蛋白也与原始红细胞的细胞膜有关。
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 the 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 primitve 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.