Abstract: The glutamate receptor agonist N‐methyl‐D‐aspartate (NMDA) stimulated a rapid, extracellular Ca2+‐dependent conversion of (3H)arginine to (3H)citrulline in primary cultures of cerebellar granule cells, indicating receptor‐mediated activation of nitric oxide (NO) synthase. The NMDA‐induced formation of (3H)citrulline reached a plateau within 10 min. Subsequent addition of unlabeled l‐arginine resulted in the disappearance of 3H from the citrulline pool, indicating a persistent activation of NO synthase after NMDA receptor stimulation. Glutamate, NMDA, and kainate, but not quisqualate, stimulated both the conversion of (3H)arginine to (3H)citrulline and cyclic GMP accumulation in a dose‐dependent manner. Glutamate and NMDA showed similar potencies for the stimulation of (3H)citrulline formation and cyclic GMP synthesis, respectively, whereas kainate was more potent at inducing cyclic GMP accumulation than at stimulating (3H)citrulline formation. Both the (3H)arginine to (3H)citrulline conversion and cyclic GMP synthesis stimulated by NMDA were inhibited by the NMDA receptor antagonist MK‐801 and by the inhibitors of NO synthase, NG‐monomethyl‐L‐arginine (MeArg) and NG‐nitro‐L‐arginine (NOArg). However, MeArg, in contrast to NOArg, also potently inhibited (3H)arginine uptake. Kainate (300 μM) stimulated 45Ca2+ influx to the same extent as 100 μM NMDA, but stimulated (3H)citrulline formation to a much lesser extent, which suggests that NO synthase is localized in subcellular compartments where the Ca2+ concentration is regulated mainly by the NMDA receptor.
Journal of Neurochemistry – Wiley
Published: Jan 1, 1992
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