Basolateral pH-sensitive K+ channels mediate membrane potential of proximal tubule cells in bullfrog kidney

The present study investigated basolateral K+ channels and their pH-sensitivity in isolated bullfrog proximal tubule cells by using the patch-clamp technique, and compared channel activity with the basolateral membrane potential (EM) and intracellular pH (pHi) monitored by using double-barreled H+-selective microelectrodes in perfused bullfrog proximal tubules. In the patch-clamp experiments, K+ channels with inward slope conductance of about 50 pS were observed in the basolateral membrane of isolated proximal tubule cells in cell-attached patches. Raising pH of the bath with HCO3(-)-free HEPES Ringer solutions from 7.7 (control) to 8.2 in the presence of an H+ ionophore, FCCP (2 micro M), enhanced channel activity to 126.4% of controls; lowering bath pH to 7.2 and to 6.7 reduced channel activity to 26.4 and to 1.7% of controls. Microelectrode experiments in bullfrog proximal tubules perfused with HCO3(-) -free HEPES Ringer solutions showed that EM and pHi in control conditions of peritubular pH 7.7 without FCCP were -52.6 mV and 7.45. Raising peritubular pH to 8.2 in the presence of FCCP (2 micro M) increased EM and pHi to -61.8 mV and 7.73; lowering it to 7. 2 and to 6.7 decreased EM and pHi to -28.6 mV and 7.25 and to -10.6 mV and 6.95. These results suggest that changes in EM in response to cellular alkalinization or acidification made by HCO3(-)-free HEPES solutions are produced primarily by changes in activity of the pH-sensitive K+ channels.