Outward K+ current in epithelial cells isolated from intermediate portion of endolymphatic sac of guinea pigs.

Abstract

Ion currents in epithelial cells isolated from the intermediate portion of endolymphatic sac (ES) in guinea pigs were investigated with the use of the whole cell patch-clamp technique. Depolarizing voltage steps from a holding potential of -60 mV induced a time- and voltage-dependent outward current, which is comparable to that of delayed rectifying K+ currents. The average resting membrane potential in the current-clamp mode was -54.8 +/- 11 mV (n = 45), which was similar to the value of zero current potential (-55.6 +/- 0.8 mV, n = 32) obtained from current-voltage (I-V) relationships of outward currents in voltage-clamp mode. The I-V relationship of the tail current exhibited a reversal potential (Erev) of -78.1 +/- 0.9 mV (n = 19) in standard external solution. The Erev of the outward current was linearly related to the logarithm of extracellular K+ concentrations. The slope was 48 mV per 10-fold change in extracellular K+ concentrations. The time constants of K+ current activation, inactivation, and K+ tail current deactivation were voltage dependent. The steady-state activation and inactivation of K+ current exhibited a sigmoidal relationship to voltage. The 50% maximal activation voltage and slope factor were -21 and 11 mV (n = 8), respectively. The 50% maximal inactivation voltage and slope factor were -45 and 13 mV (n = 7), respectively. The K+ current was blocked by externally applied 1 mM 4-aminopyridine (4-AP), 5 mM Ba2+ and 20 mM tetraethylammonium chloride (TEA). The sensitivity of the current to 4-AP and Ba2+ was higher than that to TEA. Elimination of external Ca2+ and increase of internal Ca2+ failed to significantly change the current, suggesting that the K+ current may be Ca2+ independent. The results show that epithelial cells in the intermediate portion of the ES possess a delayed-rectifier K+ current, which may be involved in membrane stability or in the ion balance between the cytosol and the extracellular environment.