Nitric Oxide Synthase Activity Endogenously Modulates NMDA Receptors

Abstract: We tested the possibility that endogenous nitric oxide synthase activity regulated NMDA receptors in primary cultured striatal neurons. We monitored NMDA-induced increase in intra-cellular Ca²⁺ levels with fura-2 ratio imaging, while nitric oxide synthase activity was either increased with l -arginihe (the natural substrate of nitric oxide synthase) or inhibited using nitro- l -arginine (a specific inhibitor of nitric oxide synthase). We found that the NMDA receptor effect was slowly but strongly diminished after an l -arginine (1 m M , 15 min) treatment ( l -arginine preincubation reduced the 100 μM NMDA-induced maximal effect by 30–50%). The l -arginine blockade of NMDA receptors was long-lasting but could be partially reversed by hemoglobin (100 μM , 10 min), which binds nitric oxide. This was not observed when the neurons were treated with l -arginine together with nitro- l -arginine. Our data strongly suggest that physiological nitric oxide synthase activity could regulate NMDA receptors.     

Reversed-phase high-performance liquid chromatography method for the analysis of nitro-arginine in rat plasma and urine

Nitro- -arginine ( -NNA) is an inhibitor of the enzyme nitric oxide synthase (NOS). We developed a simple, sensitive and reproducible reversed-phase high-performance liquid chromatographic method for detection of nitro-arginine ( - and -enantiomer) in rat plasma and urine. Samples were treated with perchloric acid, neutralized and eluted through a C₈ reversed-phase column with a mobile phase of 18.5 mM heptanesulfonic acid-10% methanok in water using theophylline as an internal standard. Plasma recovery for both isomers was complete, and the sensitivity limit was 0.5 μg/ml. This method may be used for disposition studies of -NNA in small animals.     

Stereotaxic Administration of 1-Methyl-4-Phenylpyridinium Ion (MPP+) Decreases Striatal Fructose 2,6-Bisphosphate in Rats

Abstract: The Stereotaxic administration of 1-methyl-4-phenylpyridinium ion (MPP⁺) into the neostriatum of male rats caused a lesion that resulted in a large dose-dependent loss of striatal fructose 2,6-bisphosphate; initial values were restored 5 days after the treatment. This effect was not protected by systemic administration of MK-801 or by nitroarginine. The content of hexose 6-phosphates and ATP was also reduced by MPP⁺ treatment, whereas lactate was increased. Biochemical and histological results suggested that MPP⁺ caused a nonselective cell death, followed by a pronounced astroglial response, parallel to fructose 2, 6-bisphosphate recovery. The Stereotaxic administration of rotenone showed a different time effect on fructose 2,6-bisphosphate cerebral content, with a significantly faster recovery. These results indicate that cerebral fructose 2,6-bisphosphate may be a sensitive metabolite related to brain damage caused by potent neurotoxins such as MPP⁺. On the other hand, they show that MPP⁺ acts in the brain through a quick, strong cytotoxic mechanism, which probably involves mechanisms other than mitochondria! chain blockage