In mouse striatal neurons in primary culture, the maximal increase in intracellular cyclic guanosine monophosphate level evoked by N-methyl-d-aspartic acid (NMDA) receptor activation was twice that induced by kainate, KCl and ionomycin. Quisqualate was almost inactive. All responses were mediated by nitric oxide (NO) production since they were blocked by haemoglobin (a NO scavenger) and by l-NG-monomethylarginine and l-NG-nitroarginine, the effects of both arginine analogues being reversed by an excess of l-arginine. Several results indicate that NMDA receptors stimulate a specific NO synthase activity. This specifically NMDA-activated NO synthase was blocked by nanomolar concentrations of l-NG-nitroarginine, whereas the responses evoked by other agents, including kainate, KCl and ionomycin, were only blocked by micromolar concentrations of this NO synthase inhibitor. The NMDA response could not be totally reproduced by an increase in cytosolic calcium (Ca2+) alone. In contrast, in the presence of staurosporine, an inhibitor of protein kinases C (PKC), as well as after desensitization of PKC induced by long-term treatment with the phorbol ester, phorbol-12, 13-dibutyrate, NMDA-stimulated NO production was selectively reduced, reaching the level evoked by kainate or Ca2+ increase. In conclusion, our results suggest that in striatal neurons, NMDA selectively stimulates a NO synthase activity which is inhibited by low concentrations of l-NG-nitro-arginine, through a mechanism involving PKC.