K B Walsh1, C Wang. 1. Department of Pharmacology, University of South Carolina, School of Medicine, Columbia 29208, USA.
Abstract
OBJECTIVES: The aim of this study was to determine the effects of Cl- channel blockers on the cardiac cystic fibrosis transmembrane conductance regulator (CFTR) Cl- current (ICl) and the protein kinase A-regulated L-type calcium current (PKA-ICa). METHODS: Whole-cell ICl and ICa were recorded from isolated guinea pig ventricular myocytes using the patch clamp technique during stimulation of PKA by forskolin (1 or 2 microM). RESULTS: The inhibitory effects of clofibric acid, p-chlorophenoxy propionic acid, gemfibrozil, diphenylamine-2-carboxylate (DPC), anthracene-9-carboxylate, 4,4'dinitrostilbene-2,2'-disulfonic acid and indanyloxyacetic acid 94 were examined on the two currents. Clofibric acid (1 mM), p-chlorophenoxy propionic acid (1 mM) and gemfibrozil (250 microM) produced an approximate 50% decrease in ICl, but had no effect on the PKA-ICa. Surprisingly, application of DPC (500 microM and 1 mM) and anthracene-9-carboxylate (500 microM) strongly reduced both currents. However, inhibition of the Ca2+ and Cl- channels by DPC could be differentiated in two important ways. First, increasing the pH of the external solution from 7.4 to 10.0 prevented the block of ICl by DPC, but did not attenuate the reduction in the PKA-ICa. Second, DPC inhibited the PKA-ICa in mouse atrial myocytes which lacked ICl. Neither 4,4'dinitrostilbene-2,2'-disulfonic acid (100 microM) nor indanyloxyacetic acid 94 (50 microM) caused any change in either of the guinea pig ventricular currents. CONCLUSIONS: Drugs such as DPC and anthracene-9-carboxylate which block the cardiac CFTR Cl- channel also inhibit the regulation of the L-type ICa. During beta-adrenergic stimulation, changes produced by these drugs on the cardiac action potential duration will be attributable to inhibition of both the Cl- and Ca2+ currents. Analogues of clofibric acid may serve as selective blockers of the CFTR Cl- channel that can be used to determine the physiological function of ICl in cardiac excitation.
OBJECTIVES: The aim of this study was to determine the effects of Cl- channel blockers on the cardiac cystic fibrosis transmembrane conductance regulator (CFTR) Cl- current (ICl) and the protein kinase A-regulated L-type calcium current (PKA-ICa). METHODS: Whole-cell ICl and ICa were recorded from isolated guinea pig ventricular myocytes using the patch clamp technique during stimulation of PKA by forskolin (1 or 2 microM). RESULTS: The inhibitory effects of clofibric acid, p-chlorophenoxy propionic acid, gemfibrozil, diphenylamine-2-carboxylate (DPC), anthracene-9-carboxylate, 4,4'dinitrostilbene-2,2'-disulfonic acid and indanyloxyacetic acid 94 were examined on the two currents. Clofibric acid (1 mM), p-chlorophenoxy propionic acid (1 mM) and gemfibrozil (250 microM) produced an approximate 50% decrease in ICl, but had no effect on the PKA-ICa. Surprisingly, application of DPC (500 microM and 1 mM) and anthracene-9-carboxylate (500 microM) strongly reduced both currents. However, inhibition of the Ca2+ and Cl- channels by DPC could be differentiated in two important ways. First, increasing the pH of the external solution from 7.4 to 10.0 prevented the block of ICl by DPC, but did not attenuate the reduction in the PKA-ICa. Second, DPC inhibited the PKA-ICa in mouse atrial myocytes which lacked ICl. Neither 4,4'dinitrostilbene-2,2'-disulfonic acid (100 microM) nor indanyloxyacetic acid 94 (50 microM) caused any change in either of the guinea pig ventricular currents. CONCLUSIONS: Drugs such as DPC and anthracene-9-carboxylate which block the cardiac CFTR Cl- channel also inhibit the regulation of the L-type ICa. During beta-adrenergic stimulation, changes produced by these drugs on the cardiac action potential duration will be attributable to inhibition of both the Cl- and Ca2+ currents. Analogues of clofibric acid may serve as selective blockers of the CFTR Cl- channel that can be used to determine the physiological function of ICl in cardiac excitation.
Authors: Krisztina Váczi; Bence Hegyi; Ferenc Ruzsnavszky; Kornél Kistamás; Balázs Horváth; Tamás Bányász; Péter P Nánási; Norbert Szentandrássy; János Magyar Journal: Naunyn Schmiedebergs Arch Pharmacol Date: 2014-10-26 Impact factor: 3.000
Authors: Beth Ann Murphy; Kurt A Fakira; Zhentao Song; Annie Beuve; Vanessa H Routh Journal: Am J Physiol Cell Physiol Date: 2009-07-01 Impact factor: 4.249
Authors: Dulce Figueiras-Fierro; Juan José Acevedo; Pablo Martínez-López; Jessica Escoffier; Francisco V Sepúlveda; Enrique Balderas; Gerardo Orta; Pablo E Visconti; Alberto Darszon Journal: J Cell Physiol Date: 2013-03 Impact factor: 6.384