Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates
Force and focal adhesion assembly: a close relationship studied using elastic micropatterned substrates
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DOI：
10.1038/35074532
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发表时间：
2001-05-01
影响因子：
21.3
通讯作者：
Geiger, B
中科院分区：
文献类型：
--
作者：
Balaban, NQ;Schwarz, US;Geiger, B
机械力在细胞黏附和细胞骨架组织的调节中起着重要作用。为了探索这种调节背后的分子机制，我们研究了细胞施加于基质的局部力与粘着斑组装之间的关系。我们开发了一种新方法，用于实时、高分辨率地测量细胞在单个黏附位点施加的力。这种方法将弹性体基质的微图案化与表达绿色荧光蛋白标记的纽蛋白的活细胞中粘着斑的荧光成像相结合。局部力与粘着斑的方向、总荧光强度和面积相关，表明应力恒定为5.5±2纳牛/平方微米。通过阻断肌动球蛋白的收缩性，对粘着斑的力依赖性调节动力学进行了表征，发现其时间尺度为秒级。这些结果对粘着斑对施加力的机械感觉反应的可能分子机制提出了明确的限制。
Mechanical forces play a major role in the regulation of cell adhesion and cytoskeletal organization. In order to explore the molecular mechanism underlying this regulation, we have investigated the relationship between local force applied by the cell to the substrate and the assembly of focal adhesions. A novel approach was developed for real-time, high-resolution measurements of forces applied by cells at single adhesion sites. This method combines micropatterning of elastomer substrates and fluorescence imaging of focal adhesions in live cells expressing GFP-tagged vinculin, Local forces are correlated with the orientation, total fluorescence intensity and area of the focal adhesions, indicating a constant stress of 5.5 +/- 2 nN mum-2. The dynamics of the force-dependent modulation of focal adhesions were characterized by blocking actomyosin contractility and were found to be on a time scale of seconds. The results put clear constraints on the possible molecular mechanisms for the mechanosensory response of focal adhesions to applied force.