Mapping mechanical strain of an endogenous cytoskeletal network in living endothelial cells
Mapping mechanical strain of an endogenous cytoskeletal network in living endothelial cells
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DOI：
10.1016/s0006-3495(03)75074-7
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发表时间：
2003-04-01
影响因子：
3.4
通讯作者：
Davies, PF
中科院分区：
文献类型：
--
作者：
Helmke, BP;Rosen, AB;Davies, PF
细胞机械化学信号传导的一个中心方面是在施加外源力时细胞骨架张力的变化。在这里，我们报告测量的机械应变的时空分布在中间丝细胞骨架的内皮细胞计算内源性绿色荧光蛋白（GFP）-波形蛋白的相对位移之前和之后发生的剪切应力。定量图像分析允许计算的主要值和方向的拉格朗日应变从3-D高分辨率荧光强度分布，描述中间丝的位置。在剪切应力发生后，空间局部峰值在中间丝应变重新定位。主应变的方向表明，机械拉伸引起的细胞边界。这种使用内源性报告基因进行细胞内应变作图的新方法证明了力从内腔表面转移到整个细胞。
A central aspect of cellular mechanochemical signaling is a change of cytoskeletal tension upon the imposition of exogenous forces. Here we report measurements of the spatiotemporal distribution of mechanical strain in the intermediate filament cytoskeleton of endothelial cells computed from the relative displacement of endogenous green fluorescent protein (GFP)-vimentin before and after onset of shear stress. Quantitative image analysis permitted computation of the principal values and orientations of Lagrangian strain from 3-D high-resolution fluorescence intensity distributions that described intermediate filament positions. Spatially localized peaks in intermediate filament strain were repositioned after onset of shear stress. The orientation of principal strain indicated that mechanical stretching was induced across cell boundaries. This novel approach for intracellular strain mapping using an endogenous reporter demonstrates force transfer from the lumenal surface throughout the cell.