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Am J Physiol Cell Physiol 283: C115-C125, 2002. First published February 20, 2002; doi:10.1152/ajpcell.00038.2001
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Vol. 283, Issue 1, C115-C125, July 2002

RhoA exerts a permissive effect on volume-regulated anion channels in vascular endothelial cells

Iris Carton1, Dominique Trouet1, Diane Hermans1, Holger Barth2, Klaus Aktories2, Guy Droogmans1, Nanna K. Jorgensen3, Else K. Hoffmann4, Bernd Nilius1, and Jan Eggermont1

1 Laboratory of Physiology, Katholieke Universiteit Leuven, Campus Gasthuisberg, B-3000 Leuven, Belgium; 2 Institute of Experimental and Clinical Pharmacology and Toxicology, 79104 Freiburg, Germany; 3 Department of Medical Physiology, The Panum Institute, University of Copenhagen, 2200 Copenhagen; and 4 Biochemical Department, August Krogh Institute, University of Copenhagen, 2100 Copenhagen, Denmark

Cell swelling triggers in most cell types an outwardly rectifying anion current, ICl,swell, via volume-regulated anion channels (VRACs). We have previously demonstrated in calf pulmonary artery endothelial (CPAE) cells 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 us 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 cells with constitutively active isoforms of Galpha (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 guanosine 5'-O-(3-thiotriphosphate) or C3 exoenzyme had no effect on VRACs in caveolin-1-expressing Caco-2 cells. We conclude that the Rho pathway exerts a permissive effect on VRACs in CPAE cells, i.e., swelling-induced opening of VRACs requires a functional Rho pathway, but not an activation of the Rho pathway.

cytoskeleton; actin; myosin phosphorylation; RhoA pathway


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