Force measurements in E-cadherin-mediated cell doublets reveal rapid adhesion strengthened by actin cytoskeleton remodeling through Rac and Cdc42.
Force measurements in E-cadherin-mediated cell doublets reveal rapid adhesion strengthened by actin cytoskeleton remodeling through Rac and Cdc42.
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DOI：
10.1083/jcb.200403043
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发表时间：
2004-12-20
期刊：
影响因子：
--
通讯作者：
Dufour S
中科院分区：
文献类型：
--
作者：
Chu YS;Thomas WA;Eder O;Pincet F;Perez E;Thiery JP;Dufour S
我们已经使用了一个修改后的，双移液管测定来量化钙粘蛋白依赖的细胞间粘附的强度。测量分离悬浮液中表达E-钙粘蛋白的成对细胞所需的力作为细胞间粘附的指数。分离力依赖于功能性钙粘蛋白在细胞表面的亲同性相互作用，随着接触的持续时间和钙粘蛋白水平的增加。切断钙粘蛋白和肌动蛋白细胞骨架之间的联系或破坏肌动蛋白聚合并不影响钙粘蛋白介导的粘附的启动，但可以防止其随着时间的推移而发展和变得更强。Rac和Cdc 42，Rho样的小GTP酶，被激活时，E-钙粘蛋白表达细胞在悬浮液中形成聚集。Rac或Cdc 42的显性负性形式的过度生产允许初始基于E-钙粘蛋白的粘附，但影响其后来的发展;显性活性形式彻底阻止细胞粘附。我们的研究结果强调了Rac，Cdc 42和肌动蛋白细胞骨架动力学在强细胞粘附的发展和调节中发挥的关键作用，定义为机械力。
We have used a modified, dual pipette assay to quantify the strength of cadherin-dependent cell–cell adhesion. The force required to separate E-cadherin–expressing paired cells in suspension was measured as an index of intercellular adhesion. Separation force depended on the homophilic interaction of functional cadherins at the cell surface, increasing with the duration of contact and with cadherin levels. Severing the link between cadherin and the actin cytoskeleton or disrupting actin polymerization did not affect initiation of cadherin-mediated adhesion, but prevented it from developing and becoming stronger over time. Rac and Cdc42, the Rho-like small GTPases, were activated when E-cadherin–expressing cells formed aggregates in suspension. Overproduction of the dominant negative form of Rac or Cdc42 permitted initial E-cadherin–based adhesion but affected its later development; the dominant active forms prevented cell adhesion outright. Our findings highlight the crucial roles played by Rac, Cdc42, and actin cytoskeleton dynamics in the development and regulation of strong cell adhesion, defined in terms of mechanical forces.
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