Caffeine- and carbachol-induced Cl- and cation currents in single opossum esophageal circular muscle cells.

Abstract

Cl- and cation currents may play important roles in esophageal smooth muscle membrane potential changes and contraction. We studied Ca2+ release-activated cell-shortening and membrane currents in single cells freshly dispersed from the circular muscle of the opossum esophagus using the standard patch-clamp whole cell recording method. Caffeine (10-20 microM) and carbachol (10-100 microM) shortened the single smooth muscle cells by releasing intracellular Ca2+. At a holding potential of 0 mV, spontaneous transient outward currents STOCs, representing spontaneous Ca(2+)-activated K+ currents) were recorded. Caffeine, carbachol, or ionomycin evoked large outward currents (up to 1,650 pA) and subsequently abolished STOCs. At a holding potential of -50 mV in K(+)-containing solutions, an outward current in response to the agonists was observed; in some cells, the outward current followed an inward current. In K(+)-free solutions, the agonists induced only an inward current whose reversal potential was shifted by alteration of the anion gradient but not by that of the cation. With a low-Cl- pipette solution (Cl- substituted by glucuronate or glutamate), the inward currents were dependent mainly on the external cation gradient. This cation channel was permeable to Ba2+. Inclusion of 10 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid in the pipette solution abolished all these currents. These data suggest that in the opossum esophageal circular muscle 1) Ca2+ released from the intracellular stores by caffeine and carbachol is sufficient to induce single smooth muscle cell contraction and 2) the caffeine-, carbachol-, and ionomycin-induced membrane currents consist of Ca(2+)-activated K+, Cl-, and cation conductances.