K+ currents in rabbit esophageal muscularis mucosae

Single cells were obtained from esophageal muscularis mucosae of the rabbit using enzymatic dispersion. Their electrophysiological properties were studied with both conventional whole cell and nystatin-perforated patch techniques. The latter technique was used to prevent "washout" of intracellular constituents and to maintain endogenous buffering of Ca2+. The average resting potential of these cells was -54 +/- 3.2 mV in the conventional recording and -51 +/- 4.4 mV in perforated patch recordings. In the current-clamp mode, regenerative responses were consistently observed in perforated patch recordings, but not when conventional whole cell gigaseal methods were used. Conventional whole cell voltage-clamp methods revealed outward currents on depolarization from a holding potential of -70 mV. These currents were inhibited by extracellular tetraethylammonium (TEA, 5-10 mM) and CoCl2 (4 mM), indicating that the predominant outward current is a Ca(2+)-activated K+ current. In the presence of TEA, inward Ca2+ currents were unmasked. In contrast, when the nystatin-perforated patch technique was used, depolarizations resulted in a net inward current followed by an outward current. The outward current was inhibited by CoCl2 (2 mM) and TEA (5 mM) to the same extent as in conventional recordings. A second component of K+ current was observed in both types of recordings when extracellular Ca2+ influx was abolished and also in the presence of TEA. This slowly activating persistent K+ current resembled a delayed rectifier K+ current. These studies show that the rabbit tunica muscularis mucosal cells possess voltage-activated Ca2+ and K+ channels as well as the capability to elicit action potentials.