Regulation of the GABA(A) receptor by nitric oxide in frog pituitary melanotrophs.
Nitric oxide (NO) is implicated in the regulation of various endocrine functions, but the effect of NO on GABA(A) receptor transmission has never been reported in endocrine cells. In the present study, we have investigated the effects of various agents acting on the NO transduction pathway on GABA(A) receptor function in frog pituitary melanotrophs. Histochemical studies using the NADPH-diaphorase reaction and immunohistochemical labeling with antibodies against neuronal NO synthase (nNOS) revealed that nNOS is expressed in the intermediate lobe of the pituitary and in cultured melanotrophs. Whole-cell patch-clamp recordings showed that the specific substrate of NOS L-arginine (L-Arg, 10(-4) M) or the NO donor sodium nitroprusside (10(-5) M) provoked a long-lasting inhibition of the current evoked by GABA (5 x 10(-6) M). The NOS inhibitor L-nitroarginine (10(-5) M) produced a biphasic effect, i.e. a transient decrease followed by a delayed increase of the GABA-evoked current amplitude. Similarly, the specific nNOS inhibitor 7-nitroindazole and the specific inducible NOS (iNOS) inhibitor aminoguanidine (10(-5) M each) provoked a transient depression of the current followed by a sustained potentiation. Formation of cGMP in neurointermediate lobes was enhanced by L-Arg (10(-4) M) and by the calcium-releasing agent caffeine (10(-4) M), and inhibited by the calmodulin (CaM)/Ca2+ complex blocker W7 (10(-5) M). The GABA-evoked current was potentiated by the guanylyl cyclase inhibitor ODQ (10(-8)-10(-7) M) and inhibited by the protein kinase G (PKG) activator 8pCPT-cGMP (3 x 10(-7)-3 x 10(-5) M). The present data indicate that NO, produced by a CaM/Ca2+-dependent NOS in frog melanotrophs, exerts an autocrine inhibitory effect on the GABA-evoked current. The action of NO on the GABA(A) receptor function is mediated through activation of the cGMP/PKG pathway.
Castel
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Institut National de la Santé et de la Recherche Médicale U-413, Unité Affiliée au Centre National de la Recherche Scientifique, University of Rouen, Mont-Saint-Aignan, France.
Jégou
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Tonon
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Vaudry
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D000818:Animals; D006152:Cyclic GMP / Q000378:metabolism; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism*; D009252:NADPH Dehydrogenase / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I; D018408:Patch-Clamp Techniques; D010766:Phosphorylation; D010902:Pituitary Gland / Q000166:cytology / Q000378:metabolism*; D011895:Rana ridibunda; D011963:Receptors, GABA-A / Q000378:metabolism*; D000521:alpha-MSH / Q000378:metabolism
D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D020030:Nitric Oxide Donors; D011963:Receptors, GABA-A; D009569:Nitric Oxide; D000521:alpha-MSH; D009074:Melanocyte-Stimulating Hormones; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D009252:NADPH Dehydrogenase; D006152:Cyclic GMP