Svitkina T M
Tsitologiia. 1989 Oct;31(10):1158-64.
Spreading of mouse embryo fibroblasts in the presence of cytochalasin D (1 microgram/ml) was studied using scanning electron microscopy, immunofluorescence, and electron microscopy of platinum replicas. Whereas circular lamellae were formed around the cell body during normal spreading, separate processes appeared at the cell periphery during spreading in cytochalasin-containing medium. The processes gradually elongated and branched. Cytoskeletons of fibroblasts spreading in the cytochalasin-containing medium were obtained by Triton X-100 extraction. They contained microtubules, intermediate filaments, actin "paracrystals" looking like short microfilament bundles, and patches of a meshwork-granular material. Immunogold coating of the cytoskeletons with anti-actin antibody showed that some meshwork-granular patches were decorated with gold particles, whereas the others were not. Non-actin patches were usually located on the distal ends of the processes, thus leaving behind the actin cytoskeletal components during the process growth. Another characteristic feature of this unidentified material is its usual association with the substratum and microtubules. These results suggest that the process protrusion during cell spreading in cytochalasin-containing medium may occur not due to actin polymerization as in the control cells, but due to involvement of some other non-actin cytoskeletal components. These components seem to be able to move along microtubules and to bind to the substratum.
利用扫描电子显微镜、免疫荧光以及铂复型电子显微镜技术,研究了在细胞松弛素D(1微克/毫升)存在的情况下小鼠胚胎成纤维细胞的铺展情况。在正常铺展过程中,细胞体会形成圆形薄片,而在含有细胞松弛素的培养基中铺展时,细胞周边会出现独立的突起。这些突起逐渐伸长并分支。通过Triton X-100抽提获得了在含有细胞松弛素的培养基中铺展的成纤维细胞的细胞骨架。它们包含微管、中间丝、看起来像短微丝束的肌动蛋白“副晶体”以及网状颗粒状物质斑块。用抗肌动蛋白抗体对细胞骨架进行免疫金标记显示,一些网状颗粒状斑块被金颗粒标记,而另一些则没有。非肌动蛋白斑块通常位于突起的远端,因此在突起生长过程中肌动蛋白细胞骨架成分留在了后面。这种未鉴定物质的另一个特征是它通常与基质和微管相关联。这些结果表明,在含有细胞松弛素的培养基中细胞铺展过程中的突起形成可能不是像对照细胞那样由于肌动蛋白聚合,而是由于一些其他非肌动蛋白细胞骨架成分参与其中。这些成分似乎能够沿着微管移动并与基质结合。