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Articles in PresS, published online ahead of print February 20, 2002
Am J Physiol Cell Physiol, 10.1152/ajpcell.00038.2001
Submitted on January 25, 2001
Accepted on February 18, 2002
1 Laboratory of Physiology, K.U. Leuven, Leuven, Belgium
2 Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, Freiburg, Germany
3 Department of Medical Physiology, Panum Institute, University of Copenhagen, Copenhagen, Denmark
4 Biochemical Department, August Krogh Institute, University of Copenhagen, Copenhagen, Denmark
* To whom correspondence should be addressed. E-mail: Jan.Eggermont{at}med.kuleuven.ac.be.
Cell swelling triggers in most cell types an outwardly rectifying anion current, ICl,swell, via volume-regulated anion channels (VRAC). We have previously demonstrated in calf pulmonary artery endothelial cells (CPAE) that inhibition of the Rho/Rho kinase/myosin light chain phosphorylation pathway reduces the swelling-dependent activation of ICl,swell. However, these experiments did not allow to discriminate between a direct activator role or a permissive effect. We now show that the Rho pathway did not affect VRAC activity if this pathway was activated by transfecting CPAE with constitutively active isoforms of G
13 (a Rho activating heterotrimeric G protein subunit), Rho or Rho kinase. Furthermore, biochemical and morphological analysis failed to demonstrate activation of the Rho pathway during hypotonic cell swelling. Finally, manipulating the Rho pathway with either GTP
S or C3 exoenzyme had no effect on VRAC in caveolin-1 expressing Caco-2 cells. We conclude that the Rho pathway exerts a permissive effect on VRAC in CPAE cells, i.e. swelling-induced opening of VRAC requires a functional Rho pathway, but not an activation of the Rho pathway.
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