Stress fiber reformation after ATP depletion.
Stress fiber reformation after ATP depletion.
复制标题
ATP 耗尽后应力纤维重组。
DOI：
10.1002/cm.970080204
复制
发表时间：
1987
影响因子：
--
通讯作者：
Sanger,JW
中科院分区：
文献类型：
--
作者：
GlascottJr,PA;McSorley,KM;Mittal,B;Sanger,JM;Sanger,JW
当细胞从 ATP 消耗培养基（20 mM 叠氮化钠和 10 mM 2-脱氧葡萄糖）中取出时，在应力纤维重新组装的过程中，荧光标记的重墨肌动蛋白、α-辅肌动蛋白和纽蛋白分别用于定位透化细胞和活细胞中的肌动蛋白和纽蛋白。在 80% 从 ATP 耗尽中恢复的细胞中，在去除抑制剂 5 分钟内，含有肌动蛋白、α-肌动蛋白和纽蛋白的小而分散的斑块被长而细的周期性纤维所取代。随着恢复的进展，这些新生的应力纤维变得越来越宽，直到它们达到对照细胞中应力纤维的厚度。在另外20%的细胞中，分散的斑块在逆转后5分钟内聚集，细胞中几乎所有的肌动蛋白、α-肌动蛋白和纽蛋白都集中在一个核周聚集体中，直径约为15-25μm。随着恢复的进展，所有聚集体都类似于环，其直径以约 0.5 μm/分钟的速度增加，在一些巨细胞中增长至 70 μm。随着环的尺寸增加，纤维从环向外辐射，有时跨越环的直径。在此期间细胞的形状没有改变。逆转后 1 小时，环不再存在，所有细胞都有应力纤维网络。用于定位微管蛋白和波形蛋白的间接免疫荧光技术表明，微管和中间丝不是环的组成部分，并且从反射对比光学来看，环与基底并不紧密贴合。附着斑块快速重排成核周聚集体，在细胞质中呈放射状扩散，其速度与成纤维细胞和染色体运动相同，但与其他类型的胞质内运动不同。
Flurescently labeled heavy mermoyosin, alpha‐actinin, and vinculin were used to localize actin, and vinculin, respectively, in permeabilized and living cells during the process of stress fiber reassembly, which occurred when cells were removed from ATP‐depleting medium (20 mM sodium azide and 10 mM 2‐deoxyglucose). In 80% of the cells recovering from ATP depletion, small, scattered plaques containing actin, alpha‐actinin, and vinculin were replaced by long, thin, periodic fibers within 5 minutes of removal of the inhibitors. These nascent stress fibers grew broader as recovery progressed, until they attained the thickness of stress fibers in control cells. In the other 20% of the cells, the scattered plaques aggregated within 5 minutes of reversal, and almost all the actin, alpha‐actinin, and vinculin in the cell became localized in one perinuclear aggregate, with a diameter of approximaterly 15–25 μm. As recovery progressed, all aggregates resembled rings, with diameters that increased at about 0.5 μm/minute and grew to as large as 70 μm in some giant cells. As the size of the rings increased, fibers radiated outward from them and sometimes spanned the diamater of te rings. The shape of the cells did not change during this time. By 1 hour after reversal, the rings were no longer present and all cells had networks of stress fibers. Indirect immunofluorescence techniques used to localize tubulin and vimentin indicated that microtubules and intermediate filaments were not constituents of the rings, and the rings were not closely apposed to the substrate, judging from reflection contrast optics. The rapid rearrangement of attachment plaques into a perinuclear aggregate that spreads radially in the cytoplasm occurs at the same speed as fibroblast and chromosomal movement, but is unlike other types of intracytoplasmic motility.