Mechanism for transition from initial to stable cell-cell adhesion: Kinetic analysis of E-cadherin-mediated adhesion using a quantitative adhesion assay
Mechanism for transition from initial to stable cell-cell adhesion: Kinetic analysis of E-cadherin-mediated adhesion using a quantitative adhesion assay
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DOI：
10.1083/jcb.134.2.549
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发表时间：
1996-07-01
影响因子：
7.8
通讯作者：
Nelson, WJ
中科院分区：
文献类型：
--
作者：
Angres, B;Barth, A;Nelson, WJ
基于离心力的粘附试验已被用来定量研究的动力学形成的细胞-细胞接触的糖皮质激素诱导型启动子的控制下，在小鼠成纤维细胞中的表达的E-钙粘蛋白介导的特异性。对表达最大或最小水平的E-钙粘蛋白的细胞的分析表明，E-钙粘蛋白介导的粘附强度在短时间内（半最大粘附，13-17分钟）以单一指数步骤发展。在37 ℃下，粘附强度在前20分钟内迅速增加，没有明显的滞后期。90分钟后，粘合强度达到平台。粘附的最终强度的差异与E-钙粘蛋白表达的水平相称，粘附的加强是温度依赖性的。在19 ℃下，粘附力的增强被延迟，并且随后与37 ℃下的粘附力相比以较慢的速率发展。在4 ° C下，粘附被完全抑制。粘附的加强完全依赖于功能性肌动蛋白细胞骨架，因为当细胞用细胞松弛素D处理时，粘附不会发展。总之，我们目前和以前的（麦克尼尔等人，1993. 1217-1226）的研究表明，E-钙粘蛋白介导的粘附的初始增强速率既不依赖于E-钙粘蛋白表达的量，也不依赖于膜中蛋白质向粘附位点的长程扩散。然而，最初的加强粘附依赖于温度敏感的细胞活动，可能会本地耦合集群的E-钙粘蛋白的肌动蛋白细胞骨架。
A centrifugal force-based adhesion assay has been used to quantitatively examine the kinetics of formation of cell-cell contacts mediated specifically by expression of E-cadherin under the control of a glucocorticoid-inducible promoter in mouse fibroblasts. Analysis of cells expressing maximal or minimal levels of E-cadherin showed that the strength of E-cadherin-mediated adhesion developed in a single exponential step over a short time (half-maximal adhesion, 13-17 min). At 37 degrees C, adhesion strength increased rapidly in the first 20 min without an apparent lag phase. After 90 min, adhesion strength reached a plateau. Differences in final strengths of adhesion were commensurate with the level of E-cadherin expression, Strengthening of adhesion was temperature dependent. At 19 degrees C, strengthening of adhesion was delayed and subsequently developed with a slower rate compared to adhesion at 37 degrees C. At 4 degrees C, adhesion was completely inhibited. Strengthening of adhesion was absolutely dependent on a functional actin cytoskeleton since adhesion did not develop when cells were treated with cytochalasin D. Together, our current and previous (McNeill et al., 1993. J. Cell Biol. 120:1217-1226) studies indicate that the rate of initial strengthening of E-cadherin-mediated adhesion is neither dependent on the amount of E-cadherin expressed nor on long-range protein diffusion in the membrane to the adhesion site. However, initial strengthening of adhesion is dependent on temperature-sensitive cellular activities that may locally couple clusters of E-cadherin to the actin cytoskeleton.