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Author:
Date:
2016-03-05
[Abstract] Numerous biological functions such as cytokinesis, changes in cell shape and cell migration require actomyosin-driven cellular contractility. However, the detailed mechanism of how contractile forces drive cellular processes are difficult to decipher due to the complexity of the intracellular environment. In particular, the mesoscopic description of the myosin II-dependent actin retrograde flow in cell lamellum is missing. Here, we describe a methodology for detergent extraction of cell, which preserves integrity of the actin cytoskeleton. This semi-in vitro cell model allows for the observation, using light microscopy, and quantification of changes in the actin cytoskeleton resulting from the activation of cellular contractility upon addition of ATP. This assay also allows for ...
[摘要] 许多生物学功能如细胞分裂，细胞形状和细胞迁移的变化需要肌动蛋白驱动的细胞收缩性。然而，由于细胞内环境的复杂性，收缩力如何驱动细胞过程的详细机制难以解读。特别是，细胞层中的肌球蛋白II依赖性肌动蛋白逆行流的介观描述缺失。在这里，我们描述了洗涤剂提取的细胞，保留肌动蛋白细胞骨架的完整性的方法。这种半体外细胞模型允许使用光学显微镜观察和定量在添加ATP时由于细胞收缩性的活化而导致的肌动蛋白细胞骨架中的变化。该测定还允许评估肌动蛋白相关蛋白和其他相关因子在肌动蛋白收缩活性的调节中的作用。在这里，我们证明逆行流动的着名的基于肌动蛋白的结构 - 横向弧，这是肌球蛋白IIA含有结构出现在lamellipodium-lamellum之间的边界和肌球蛋白II依赖的方式向心移动。
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