2 PATHWAYS OF CYCLIC-GMP PRODUCTION THROUGH GLUTAMATE RECEPTOR-MEDIATED NITRIC-OXIDE SYNTHESIS

The selective agonists for the metabotropic glutamate receptor,and the ionotropic non-N-methyl-D-aspartate (NMDA) glutamate receptor, (+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (ACPD) and (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA), respectively, increased the cyclic GMP (cGMP) content in cerebellar slices prepared from adult rats. The ACPD-induced rise in cGMP level was blocked by compounds known to antagonize metabotropic glutamate receptors, such as DL-2-amino-3-phosphonopropionic acid and L-2-amino-4-phosphonobutyric acid, but not by ionotropic glutamate receptor antagonists, D-2-amino-5-phosphonovaleric acid and 6 - cyano - 7 - nitroquinoxaline - 2, 3 - dione (CNQX), whereas the AMPA-induced rise in cGMP level was suppressed by CNQX. Both rises in cGMP level involved nitric oxide synthase (NOS), because N(G)-methyl-L-arginine (NMLA), an inhibitor of NOS, blocked both cGMP level rises, and excess L-arginine reversed the effect of NMLA. After lithium chloride treatment, which could exhaust phosphatidylinositol phosphates, ACPD no longer increased cGMP levels, whereas AMPA was still effective. In a calcium-free medium, ACPD still induced a rise in cGMP level, whereas AMPA did not. When the molecular layer was isolated to determine-the cGMP content separately from that in the rest of the cerebellar cortex, it was found that ACPD raised the cGMP level mainly in the molecular layer, whereas AMPA raised it in both sections. These results suggest that ACPD enhances the cGMP level through activation of NOS independently of extracellular calcium, most likely by calcium release from intracellular stores triggered by metabotropic glutamate receptors linked to phosphoinositide breakdown, whereas AMPA activates the enzyme by calcium entry from the extracellular space triggered by ionotropic non-NMDA glutamate receptors.