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Published online 13 December 2004. doi:10.1083/jcb.200403043
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 6, 1183-1194
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Article

Force measurements in E-cadherin–mediated cell doublets reveal rapid adhesion strengthened by actin cytoskeleton remodeling through Rac and Cdc42



Yeh-Shiu Chu1, William A. Thomas3, Olivier Eder1, Frederic Pincet2, Eric Perez2, Jean Paul Thiery1, and Sylvie Dufour1

1 UMR144 Centre National de la Recherche Scientifique (CNRS)-Institut Curie, 75248 Paris Cedex 05, France
2 UMR8550 CNRS-ENS, 75248 Paris Cedex 05, France
3 Department of Natural Sciences, Colby-Sawyer College, New London, NH 03257

Correspondence to Sylvie Dufour: Sylvie.Dufour{at}curie.fr

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.

J.P. Thiery and S. Dufour were co-principal investigators.

Abbreviations used in this paper: {alpha}-cat, {alpha}-catenin; ß-cat, ß-catenin; BFA, brefeldin A; Ecad-{Delta}cyto, E-cadherin lacking the cytoplasmic domain; Jasp, Jasplakinolide; LatB, Latrunculin B; SF, separation force; TC, trypsin-calcium.


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