Dynamic assessment of fibroblast mechanical activity during Rac-induced cell spreading in 3-D culture.
Dynamic assessment of fibroblast mechanical activity during Rac-induced cell spreading in 3-D culture.
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DOI：
10.1002/jcp.21487
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发表时间：
2008-10
影响因子：
5.6
通讯作者：
Vishwanath, Mridula
中科院分区：
文献类型：
--
作者：
Petroll, W. Matthew;Ma, Lisha;Kim, Areum;Ly, Linda;Vishwanath, Mridula
本研究的目的是确定RAC激活对三维胶原基质内成纤维细胞的形态和亚细胞力学影响。将角膜成纤维细胞低密度接种于100μm厚的胶原纤维基质内，在无血清培养液中培养1~2d。然后使用Nomarski DIC进行时间推移成像。驯化一段时间后，将灌流切换到含有PDGF的培养液中。在一些实验中，分别使用Y-27632或Blebbistatin来抑制Rho-Kinase(ROCK)或肌球蛋白II。PDGF激活RAC并诱导细胞扩散，导致细胞长度、细胞面积和伪足突起数目增加。牵引力是通过延长伪足而产生的，如向心性移位和胶原纤维的重排所示。有趣的是，伪足延伸和局部胶原纤维重新排列的模式高度依赖于纤维在前缘的初始取向。在ROCK或肌球蛋白II抑制后，观察到明显的ECM松弛，但在伪足尖端继续检测到小的胶原纤维移位。综上所述，这些数据表明，在RAC诱导的细胞在3-D基质内扩散的过程中，角膜成纤维细胞的力分布从中心向外围转移。在细胞在3-D胶原基质内扩散时，ROCK介导了基于肌球蛋白II的大牵引力的产生，然而，在延伸伪足的尖端可以产生残余力，这两种力都不依赖于ROCK和肌球蛋白II。
The goal of this study was to determine the morphological and sub-cellular mechanical effects of Rac activation on fibroblasts within 3-D collagen matrices. Corneal fibroblasts were plated at low density inside 100 μm thick fibrillar collagen matrices and cultured for 1 to 2 days in serum-free media. Time-lapse imaging was then performed using Nomarski DIC. After an acclimation period, perfusion was switched to media containing PDGF. In some experiments, Y-27632 or blebbistatin were used to inhibit Rho-kinase (ROCK) or myosin II, respectively. PDGF activated Rac and induced cell spreading, which resulted in an increase in cell length, cell area, and the number of pseudopodial processes. Tractional forces were generated by extending pseudopodia, as indicated by centripetal displacement and realignment of collagen fibrils. Interestingly, the pattern of pseudopodial extension and local collagen fibril realignment was highly dependent upon the initial orientation of fibrils at the leading edge. Following ROCK or myosin II inhibition, significant ECM relaxation was observed, but small displacements of collagen fibrils continued to be detected at the tips of pseudopodia. Taken together, the data suggests that during Rac-induced cell spreading within 3-D matrices, there is a shift in the distribution of forces from the center to the periphery of corneal fibroblasts. ROCK mediates the generation of large myosin II-based tractional forces during cell spreading within 3-D collagen matrices, however residual forces can be generated at the tips of extending pseudopodia that are both ROCK and myosin II-independent.
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影响因子：
56.9
作者：
Cukierman, E;Pankov, R;Yamada, KM
通讯作者：
Yamada, KM
影响因子：
3.7
作者：
Grinnell, F;Rocha, LB;Jiang, HM
通讯作者：
Jiang, HM
影响因子：
3.4
作者：
Bohnet, S;Ananthakrishnan, R;Verkhovsky, AB
通讯作者：
Verkhovsky, AB
Rho-kinase in Vivo的肌球蛋白磷酸酶的肌球蛋白结合亚基（MB）的磷酸化。
DOI：
10.1083/jcb.147.5.1023
发表时间：
1999-11-29
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kawano Y;Fukata Y;Oshiro N;Amano M;Nakamura T;Ito M;Matsumura F;Inagaki M;Kaibuchi K
通讯作者：
Kaibuchi K
DOI：
10.1083/jcb.200204153
发表时间：
2002-11-25
期刊：
The Journal of cell biology
影响因子：
--
作者：
Galbraith CG;Yamada KM;Sheetz MP
通讯作者：
Sheetz MP