Further analysis of ATP-mediated activation of K+ channels in renal epithelioid Madin Darby canine kidney (MDCK) cells

Abstract

ATP activates K+ channels by increasing intracellular calcium activity in Madin Darby canine kidney (MDCK) cells. The present study has been performed to test for the involvement of G-proteins and of protein kinase C in the intracellular transmission of these effects. To this end, the effect of ATP on intracellular calcium and K+ channel activity has been studied in cells pretreated with the phorbol ester 12-O-tetradecanoyl-phorbol 13-acetate (TPA) and/or pertussis toxin. The ATP-induced increase of intracellular calcium is not significantly affected by pretreatment with pertussis toxin, is significantly blunted by pretreatment with TPA and is abolished by pretreatment with both pertussis toxin and the phorbol ester. The ATP activation of K+ channels is similarly blunted by pretreatment with TPA, but is not abolished by pretreatment with both the phorbol ester and pertussis toxin. Furthermore, the ATP-induced hyperpolarization is not abolished in cells pretreated with both pertussis toxin and TPA. In those cells, ATP may activate K+ channels by calcium-dependent mechanisms or lead to localized increases of intracellular calcium sufficient to activate the K+ channels but escaping detection with fura-2 fluorescence.