Ca2+-and voltage activated K+ channel in apical cell membrane of gallbladder epithelium fromTriturus

Abstract

The presence of Ca²⁺- and voltage-activated K⁺ channels was directly demonstrated in the apical cell membrane of gallbladder epithelium by patch-clamp single-channel current recording. In K⁺-depolarized epithelial cells, negative pipette potentials induced outward current steps when the patch-pipette was filled with Na⁺-rich solution and these current steps were not affected by the presence or absence of Cl⁻. When K⁺-rich solution was in the pipette and K⁺-depolarized cells were examined, the current-voltage relations were linear with a single-channel conductance of 140 pS and polarity was reversed at 0 mV. In excised inside-out membrane patches, raising the free Ca²⁺ concentration of the medium facing the inner side of the membrane from 10⁻⁷ to 10⁻⁶ M evoked a marked increase in open state probability of the channels without affecting the elementary current steps. This suggests that intracellular Ca²⁺ as a second messenger plays a crucial role in the regulatory mechanism of the membrane potential by modulating the high-conductance apical K⁺ channels.