Parallel intermediate conductance K⁺ and Cl^- channel activity mediates electroneutral K⁺ exit across basolateral...

Transepithelial K⁺ absorption requires apical K⁺ uptake and basolateral K⁺ exit. In the colon, apical H⁺-K⁺-ATPase mediates cellular K⁺ uptake, and it has been suggested that electroneutral basolateral K⁺ exit reflects K⁺-Cl^- cotransporter-1 (KCC1) operating in parallel with K⁺ and Cl^- channels. The present study was designed to identify basolateral transporter(s) responsible for K⁺ exit in rat distal colon. Active K⁺ absorption was determined by measuring ⁸⁶Rb⁺ (K⁺ surrogate) fluxes across colonic epithelia under voltage-clamp conditions. With zero Cl^- in the mucosal solution, net K⁺ absorption was reduced by 38%, indicating that K⁺ absorption was partially Cl^--dependent. Serosal addition of DIOA (KCC1 inhibitor) or Ba²⁺ (nonspecific K⁺ channel blocker) inhibited net K⁺ absorption by 21% or 61%, respectively, suggesting that both KCC1 and K⁺ channels contribute to basolateral K⁺ exit. Clotrimazole and TRAM34 (IK channel blockers) added serosally inhibited net K⁺ absorption, pointing to the involvement of IK channels in basolateral K⁺ exit. GaTx2 (CLC2 blocker) added serosally also inhibited net K⁺ absorption, suggesting that CLC2-mediated Cl^- exit accompanies IK channel-mediated K⁺ exit across the basolateral membrane. Net K⁺ absorption was not inhibited by serosal addition of either IbTX (BK channel blocker), apamin (SK channel blocker), chromanol 293B (K_V7 channel blocker), or CFTR_inh172 (CFTR blocker). Immunofluorescence studies confirmed basolateral membrane colocalization of CLC2-like proteins and Na⁺-K⁺-ATPase α-subunits. We conclude that active K⁺ absorption in rat distal colon involves electroneutral basolateral K⁺ exit, which may reflect IK and CLC2 channels operating in parallel.NEW & NOTEWORTHY This study demonstrates that during active electroneutral K⁺ absorption in rat distal colon, K⁺ exit across the basolateral membrane mainly reflects intermediate conductance K⁺ channels operating in conjunction with chloride channel 2, with a smaller, but significant, contribution from K⁺-Cl^- cotransporter-1 (KCC1) activity.