rhoa gtpase switch controls cx43-hemichannel activity through the contractile systemrhoa gtpase cx43-hemichannel活动通过收缩系统开关控制.pdfVIP

RhoA GTPase Switch Controls Cx43-Hemichannel Activity through the Contractile System 1 . 1. ´ 1 1 2 Raf Ponsaerts * , Catheleyne D’hondt , Frederic Hertens , Jan B. Parys , Luc Leybaert , 1 1 1 Johan Vereecke , Bernard Himpens , Geert Bultynck * 1 Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine, Campus Gasthuisberg O/N-1, Faculty of Medicine, KU Leuven, Leuven, Belgium, 2 Department of Basic Medical Sciences, Physiology Group, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium Abstract ATP-dependent paracrine signaling, mediated via the release of ATP through plasma membrane-embedded hemichannels of the connexin family, coordinates a synchronized response between neighboring cells. Connexin 43 (Cx43) hemichannels that are present in the plasma membrane need to be tightly regulated to ensure cell viability. In monolayers of bovine corneal endothelial cells (BCEC),Cx43-mediated ATP release is strongly inhibited when the cells are treated with inflammatory mediators, in particular thrombin and histamine. In this study we investigated the involvement of RhoA activation in the inhibition of hemichannel-mediated ATP release in BCEC. We found that RhoA activation occurs rapidly and transiently upon thrombin treatment of BCEC. The RhoA activity correlated with the onset of actomyosin contractility that is involved in the inhibition of Cx43 hemichannels. RhoA activation and inhibition of Cx43-hemichannel activity were both prevented by pre-treatment of the cells with C3-toxin as well as knock down of RhoA by siRNA. These findings provide evide