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.
Derangeon
Mickaël
M
Institut de Physiologie et Biologie Cellulaires, Université de Poitiers, F-86022 Poitiers, France.
Bourmeyster
Nicolas
N
Plaisance
Isabelle
I
Pinet-Charvet
Caroline
C
Chen
Qian
Q
Duthe
Fabien
F
Popoff
Michel R
MR
Sarrouilhe
Denis
D
Hervé
Jean-Claude
JC
D036002:ADP Ribose Transferases / Q000494:pharmacology; D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology; D001905:Botulinum Toxins / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology*; D018031:Connexin 43 / Q000378:metabolism*; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D029968:Escherichia coli Proteins / Q000494:pharmacology; D017629:Gap Junctions / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010590:Phalloidine / Q000494:pharmacology; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects / Q000502:physiology; D011042:Poisons / Q000494:pharmacology; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D062826:Zonula Occludens-1 Protein; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D000199:Actins; D001427:Bacterial Toxins; D018031:Connexin 43; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D010750:Phosphoproteins; D011042:Poisons; C569044:Tjp1 protein, rat; D062826:Zonula Occludens-1 Protein; C060296:cytotoxic necrotizing factor type 1; D010590:Phalloidine; D015638:Cytochalasin D; D000255:Adenosine Triphosphate; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; D020742:rhoA GTP-Binding Protein