Maxi K+ channels in the basolateral membrane of the exocrine frog-skin gland regulated by intracellular calcium and pH

Abstract

With the single-channel patch-clamp technique we have identified Ca2+-sensitive, high-conductance (maxi) K+ channels in the basolateral membrane (BLM) of exocrine gland cells in frog skin, Under resting conditions, maxi K+ channels were normally quiescent, but they were activated by muscarinic agonists or by high serosal K+. In excised inside-out patches and with symmetrical 140 mmol/l K+, single-channel conductance was 200 pS and the channel exhibited a high selectivity for K+ over Na+. Depolarization of the BLM increased maxi K+ channel activity. Increasing cytosolic free Ca2+ (by addition of 100 nmol/l thapsigargin to the bathing solution of cell-attached patches also increased channel activity, whereas thapsigargin had no effect when added to excised inside-out patches, An increase in cytosolic free Ca2+ directly activated channel activity in a voltage-dependent manner, Maxi K+ channel activity was sensitive to changes in intracellular pH, with maximal activity at pH 7.4 and decreasing activities following acidification and alkalinization. Maxi K+ channel outward current was reversibly blocked by micromolar concentrations of Ba2+ from the cytosolic and extracellular site, and was irreversibly blocked by micromolar concentrations of charybdotoxin and kaliotoxin from the extracellular site in outside-out patches.