DCPIB is a novel selective blocker of ICI,swell and prevents swelling-induced shortening of guinea-pig atrial action potential duration

Abstract

1. We identified the ethacrynic-acid derivative DCPIB as a potent inhibitor of ICl,swell, which blocks native ICl,sweell of calf bovine pulmonary artery endothelial (CPAE) cells with an IC50 of 4.1 μM. Similarly, 10 μM DCPIB almost completely inhibited the swelling-induced chloride conductance in Xenopus oocytes and in guinea-pig atrial cardiomyocytes. Block of ICl,swell by DCPIB was fully reversible and voltage independent. 2. DCPIB (10 μM) showed selectivity for ICl,swell and had no significant inhibitory effects on ICl,Ca in CPAE cells, on chloride currents elicited by several members of the CLC-chloride channel family or on the human cystic fibrosis transmembrane conductance regulator (hCFTR) after heterologous expression in Xenopus oocytes, DCPIB (10 μM) also showed no significant inhibition of several native anion and cation currents of guinea pig heart like ICl,PKA, IKr, IKs, IKl, INa and ICa. 3. In all atrial cardiomyocytes (n = 7), osmotic swelling produced an increase in chloride current and a strong shortening of the action potential duration (APD). Both swelling-induced chloride conductance and AP shortening were inhibited by treatment of swollen cells with DCPIB (10 μM). In agreement with the selectivity for ICl,swell, DCPIB did not affect atrial APD under isoosmotic conditions. 4. Preincubation of atrial cardiomyocytes with DCPIB (10 μM) completely prevented both the swelling-induced chloride currents and the AP shortening but not the hypotonic cell swelling. 5. We conclude that swelling-induced AP shortening in isolated atrial cells is mainly caused by activation of ICl,swell. DCPIB therefore is a valuable pharmacological tool to study the role of ICl,swell in cardiac excitability under pathophysiological conditions leading to cell swelling.