15 research outputs found
RhoA GTPase and F-actin Dynamically Regulate the Permeability of Cx43-made Channels in Rat Cardiac Myocytes*
Gap junctions are clusters of transmembrane channels allowing a passive
diffusion of ions and small molecules between adjacent cells. Connexin43, the
main channel-forming protein expressed in ventricular myocytes, can associate
with zonula occludens-1, a scaffolding protein linked to the actin
cytoskeleton and to signal transduction molecules. The possible influence of
Rho GTPases, major regulators of cellular junctions and of the actin
cytoskeleton, in the modulation of gap junctional intercellular communication
(GJIC) was examined. The activation of RhoA by cytoxic necrotizing factor 1
markedly enhanced GJIC, whereas its specific inhibition by the Clostridium
botulinum C3 exoenzyme significantly reduced it. RhoA activity affects
GJIC without major cellular redistribution of junctional plaques or changes in
the Cx43 phosphorylation pattern. As these GTPases frequently act via the
cortical cytoskeleton, the importance of F-actin in the modulation of GJIC was
investigated by means of agents interfering with actin polymerization.
Cytoskeleton stabilization by phalloidin slowed down the kinetics of channel
rundown in the absence of ATP, whereas its disruption by cytochalasin D
rapidly and markedly reduced GJIC despite ATP presence. Cytoskeleton
stabilization by phalloidin markedly reduced the consequences of RhoA
activation or inactivation. This mechanism appears to be the first described
capable to both up- or down-regulate GJIC through RhoA activation or,
conversely, inhibition. The inhibition of Rho downstream kinase effectors had
no effect on GJIC. The present results provide further insight into the gating
and regulation of junctional channels and identify a new downstream target for
the small G-protein RhoA