Possible role of phosphatidylinositol 4,5, bisphosphate in luteinizing hormone releasing hormone‐mediated M‐current inhibition in bullfrog sympathetic neurons

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Abstract
Luteinizing hormone releasing hormone (LHRH) is a physiological modulator of neuronal excitability in bullfrog sympathetic ganglia (BFSG). Actions of LHRH involve suppression of the noninactivating, voltage-dependent M-type K+ channel conductance (gM). We found, using whole-cell recordings from these neurons, that LHRH-induced suppression of gM was attenuated by the phospholipase C (PLC) inhibitor U73122 (10 microM) but not by the inactive isomer U73343 (10 microM). Buffering internal Ca2+ to 117 nM with intracellular 20 mM BAPTA + 8 mM Ca2+ or to < 10 nM with intracellular 20 mM BAPTA + 0.4 mM Ca2+ did not attenuate LHRH-induced gM suppression. Suppression of gM by LHRH was not antagonized by the inositol 1,4,5 trisphosphate (InsP3) receptor antagonist heparin (approximately 300 microM). Preventing phosphatidylinositol-4,5-bisphosphate (PIP2) synthesis by blocking phosphatidylinositol-4-kinase with wortmannin (10 microM) or with the nonhydrolysable ATP analogue AMP-PNP (3 mM) prolonged recovery of LHRH-induced gM suppression. This effect was not produced by blocking phosphatidyl inositol-3-kinase with LY294002 (10 microM). Rundown of gM was attenuated when cells were dialysed with 240 microM di-octanoyl PIP2 or 240 microM di-octanoyl phosphatidylinositol-3,4,5-trisphosphate (PIP3) but not with 240 microM di-octanoyl phosphatidylcholine. LHRH-induced gM suppression was competitively antagonized by dialysis with 240 microM di-octanoyl PIP2, but not with di-octanoyl phosphatidylcholine. These results would be expected if LHRH-induced gM suppression reflects a PLC-mediated decrease in plasma membrane PIP2 levels.