Stimulation of A2a adenosine receptor abolishes the inhibitory effectof arachidonic acid on the basolateral 50-ps K channel in the thick ascending limb

Wang M, Sui H, Li W, Wang J, Liu Y, Gu L, Wang W-H, Gu R. Stimulation of A(2a) adenosine receptor abolishes the inhibitory effect of arachidonic acid on the basolateral 50-pS K channel in the thick ascending limb. Am J Physiol Renal Physiol 300: F906-F913, 2011. First published January 5, 2011; doi:10.1152/ajprenal.00617.2010.-The basolateral 50-pS K channels are stimulated by a cAMP-dependent pathway and inhibited by cytochrome P-450-omega-hydroxylase-dependent metabolism of arachidonic acid (AA) in the rat thick ascending limb (TAL). We now used the patch-clamp technique to examine whether stimulation of adenosine A(2a) receptor modulates the inhibitory effect of AA on the basolateral 50-pS K channels in the medullary TAL. Stimulation of adenosine A(2a) receptor with CGS-21680 or inhibition of phospholipase A(2) (PLA(2)) with AACOCF3 increased the 50-pS K channel activity in the TAL. Western blot demonstrated that application of CGS-21680 decreased the phosphorylation of PLA(2) at serine residue 505, an indication of inhibiting PLA(2) activity. In the presence of CGS-21680, inhibition of PLA(2) had no further effect on the basolateral 50-pS K channels. The possibility that CGS-21680-induced stimulation of the basolateral 50-pS K channels was partially achieved by inhibition of PLA(2) in the TAL was also supported by the observation that CGS-21680 had no additional effect in the presence of AACOCF3. Moreover, stimulation of adenosine A(2a) receptor with CGS-21680 also abolished the inhibitory effect of AA and 20-hydroxyeicosatetraenoic acid (20-HETE) on the 50-pS K channels. The effect of CGS-21680 on AA and 20-HETE-mediated inhibition of the 50-pS K channels was mediated by cAMP because application of membrane-permeable cAMP analog, dibutyryl-cAMP, not only increased the 50-pS K channel activity but also abolished the inhibitory effect of AA and 20-HETE. We conclude that stimulation of adenosine A(2a) receptor increased the 50-pS K channel activity in the TAL, an effect that is achieved by suppression of PLA(2) activity and 20-HETE-induced inhibition.