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Received March 25, 2004
Revised April 15, 2004
Accepted after revision May 19, 2004
1 Kyushu University, Graduate School of Medical Sciences
* To whom correspondence should be addressed. E-mail: moike{at}pharmaco.med.kyushu-u.ac.jp.
We have investigated the cellular mechanisms of mechanical stress-induced immediate responses in human umbilical vein endothelial cells (HUVECs). Hypotonic stress (HTS) induced ATP release, which evoked Ca2+ transient, and actin reorganization in a few minutes in HUVECs. Disruption of actin cytoskeleton did not suppress HTS-induced ATP release, and the inhibition of ATP-mediated Ca2+ response did not affect actin reorganization, thereby indicating that these two responses are not interrelated. ATP release and actin reorganization were also induced by lysophosphatidic acid (LPA). HTS and LPA induced membrane translocation of RhoA, a hallmark of its activation, and tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin. Tyrosine kinase inhibitors (herbimycin A or tyrphostin 46) inhibited both HTS- and LPA-induced ATP release and actin reorganization, but did not affect RhoA activation. In contrast, Rho-kinase inhibitor (Y27632) inhibited all of the HTS- and LPA-induced responses. These results indicate that the activation of RhoA/Rho-kinase pathway followed by tyrosine phosphorylation of FAK and paxillin leads to ATP release and actin reorganization in HUVECs. Furthermore, the fact that HTS and LPA evoke completely same intracellular signals and responses suggests that even these immediate mechanosensitive responses are actually not mechanical stress-specific.
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