PIP(2) Mediated Inhibition of TREK Potassium Currents by Bradykinin in Mouse Sympathetic Neurons

Bradykinin (BK), a hormone inducing pain and inflammation, is known to inhibit potassium M-currents (I(M)) and to increase the excitability of the superior cervical ganglion (SCG) neurons by activating the Ca(2+)-calmodulin pathway. M-current is also reduced by muscarinic agonists through the depletion of membrane phosphatidylinositol 4,5-biphosphate (PIP(2)). Similarly, the activation of muscarinic receptors inhibits the current through two-pore domain potassium channels (K2P) of the “Tandem of pore-domains in a Weakly Inward rectifying K(+) channel (TWIK)-related channels” (TREK) subfamily by reducing PIP(2) in mouse SCG neurons (mSCG). The aim of this work was to test and characterize the modulation of TREK channels by bradykinin. We used the perforated-patch technique to investigate riluzole (RIL) activated currents in voltage- and current-clamp experiments. RIL is a drug used in the palliative treatment of amyotrophic lateral sclerosis and, in addition to blocking voltage-dependent sodium channels, it also selectively activates the K2P channels of the TREK subfamily. A cell-attached patch-clamp was also used to investigate TREK-2 single channel currents. We report here that BK reduces spike frequency adaptation (SFA), inhibits the riluzole-activated current (I(RIL)), which flows mainly through TREK-2 channels, by about 45%, and reduces the open probability of identified single TREK-2 channels in cultured mSCG cells. The effect of BK on I(RIL) was precluded by the bradykinin receptor (B(2)R) antagonist HOE-140 (d-Arg-[Hyp(3), Thi(5), d-Tic(7), Oic(8)]BK) but also by diC(8)PIP(2) which prevents PIP(2) depletion when phospholipase C (PLC) is activated. On the contrary, antagonizing inositol triphosphate receptors (IP(3)R) using 2-aminoethoxydiphenylborane (2-APB) or inhibiting protein kinase C (PKC) with bisindolylmaleimide did not affect the inhibition of I(RIL) by BK. In conclusion, bradykinin inhibits TREK-2 channels through the activation of B(2)Rs resulting in PIP(2) depletion, much like we have demonstrated for muscarinic agonists. This mechanism implies that TREK channels must be relevant for the capture of information about pain and visceral inflammation.