ATP-sensitive K⁺ channels maintain resting membrane potential in interstitial cells of Cajal from the mouse colon

To investigate the role of ATP-sensitive K⁺(K_ATP) channels on pacemaker activity in interstitial cells of Cajal (ICC), whole-cell patch clamping, RT-PCR, and intracellular Ca²⁺([Ca²⁺]_i) imaging were performed in cultured colonic ICC. Pinacidil (a K⁺ channel opener) hyperpolarized the membrane and inhibited the generation of pacemaker potential, and this effect was reversed by glibenclamide (a K_ATP channel blocker). RT-PCR showed that K_ir 6.1 and SUR2B were expressed in Ano-1 positive colonic ICC. Glibenclamide depolarized the membrane and increased pacemaker potential frequency. However, 5-hydroxydecanoic acid (a mitochondrial K_ATP channel blocker) had no effects on pacemaker potentials. Phorbol 12-myristate 13-acetate (PMA; a protein kinase C activator) blocked the pinacidil-induced effects, and PMA alone depolarized the membrane and increased pacemaker potential frequency. Cell-permeable 8-bromo-cyclic AMP also increased pacemaker potential frequency. Recordings of spontaneous intracellular Ca²⁺([Ca²⁺]_i) oscillations showed that glibenclamide increased the frequency of [Ca²⁺]_i oscillations. In small intestinal ICC, glibenclamide alone did not alter the generation of pacemaker potentials, and K_ir 6.2 and SUR2B were expressed in Ano-1 positive ICC. Therefore, K_ATP channels in colonic ICC are activated in resting state and play an important role in maintaining resting membrane potential.