Mechanism for nitric oxide's enhancement of NMDA-stimulated [ 3H]norepinephrine release from rat hippocampal slices

Abstract

Previous studies in our laboratory have shown that nitric oxide (NO) gas enhances NMDA-stimulated release of preloaded tritiated norepinephrine ([ 3H]NA) from rat brain slices in a dose-dependent, oxygen-sensitive, and cyclic GMP-independent manner. In this study we have attempted to determine the mechanism for the enhancement of neurotransmitter release seen with NO. NO-enhanced transmitter release was not due to buffer acidification or generation of NO degradation products, since reducing buffer pH below 7.3 inhibited NMDA-stimulated [ 3H]NA release and nitrite or nitrate ions (3–100 μM) had no significant effect on release. Carbon monoxide (CO, 10–300 μM), another diatomic gas with properties similar to NO including heme binding and guanylate cyclase activation, had no significant effect on depolarization-induced [ 3H]NA release. The NO effect was probably not due to mono-ADP-ribosylation of cellular proteins, since the ADP-ribosyltransferase (ADPRT) inhibitors nicotinamide (10 μM-10 μM) and luminol (1 μM-1 mM) did not diminish the enhancement of transmitter release seen with NO. The NA reuptake inhibitor desmesthylimipramine (DMI, 10 nM-10 μM) neither mimicked nor blocked the effect of NO, suggesting that NO was not acting via inhibition or reversal of the NA transporter. Similar to NO, the metabolic inhibitors sodium azide (NaN 3, 0.1–3 mM), potassium cyanide (KCN, 0.1–3 mM), and 2,4-dinitrophenol (2,4-DNP, 10–300 μM) also dose-dependently enhanced NMDA-stimulated [ 3H]NA release. These results suggest that NO may enhance neurotransmitter release by inhibiting cellular respiration and perhaps ultimately via altering calcium homeostasis.