Excitotoxic action of NMDA agonists on nigrostriatal dopaminergic neurons: modulation by inhibition of nitric oxide synthesis

Focal infusions of N-methyl-d-aspartate (NMDA) or an endogenous NMDA agonist, quinolinic acid (QUIN), into the substantia nigra pars compacta (SNc) of adult Sprague-Dawley rats resulted in a dose-dependent depletion of ipsilateral striatal tyrosine hydroxylase (TH) activity, a biochemical marker for dopaminergic neurons. To assess the intermediary role of nitric oxide in the neurotoxicity elicited by these toxins, their action was tested in animals treated with N^ω-nitro-l-arginine methyl ester (L-NAME). Systemic injections (2 injections; 8 h apart) of L-NAME (100, 150 and 250 mg/kg) produced a dose-related inhibition of cerebellar nitric oxide synthase (NOS) activity. The time-course of cerebellar NOS inhibition following L-NAME (250 mg/kg) was rapid in onset and lasted for at least 24 h following the second injection. An L-NAME treatment regimen of 250 mg/kg, with the second injection given 24 h prior to assessment of NOS activity, produced an 87 and 91% inhibition of cerebellar and nigral NOS activity, respectively. Intranigral infusion of 40 and 60 nmol QUIN reduced ipsilateral striatal TH activity by 62 and 75%, respectively. However, 40 and 60 nmol QUIN infusions into animals pretreated with L-NAME (250 mg/kg) reduced striatal TH activity by 83 and 96%, respectively. Intranigral infusion of 15 and 30 nmol NMDA produced a 48 and 77% decrease in striatal TH activity, respectively, whereas the same doses of NMDA given to animals pretreated with L-NAME (250 mg/kg) resulted in a 59 and 88% decrease in TH activity. Thus, both QUIN and NMDA toxicity was enhanced following L-NAME pretreatment. The destruction of the nigrostriatal pathway was verified using TH immunocytochemistry of the SNc. It was also observed that a low dose of L-NAME (1.5 mg/kg), previously shown to be neuroprotective in cerebral ischemic damage, did not influence NMDA (15 nmol) neurotoxicity. The results of this study show that extensive inhibition of NOS activity enhances NMDA receptor-mediated excitoxicity.