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This study was designed to characterize nitric oxide (NO) production and anoxic depolarization in the rat hippocampus during transient forebrain ischemia using two NO synthase (NOS) inhibitors, L-N(5)-(1-iminoethyl)ornithine (L-NIO), a relatively selective endothelial NOS (eNOS) inhibitor, and 7-nitroindazole, a relatively selective neuronal NOS (nNOS) inhibitor, and an NO scavenger, [2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide] (carboxy-PTIO). We measured the mean arterial blood pressure, hippocampal blood flow, NO concentration and direct current potential before, during and after transient forebrain ischemia, which was induced by 4-vessel occlusion for 10 min. Saline, L-NIO (20 mg/kg), 7-nitroindazole (25 mg/kg), L-NIO (20 mg/kg)+7-nitroindazole (25 mg/kg) or carboxy-PTIO (1 mg/kg) was administered intraperitoneally 20 min before the onset of ischemia. We observed early and sharp NO production in the hippocampus during ischemia in the saline group. This NO increase during ischemia was significantly reduced by L-NIO (20 mg/kg)+7-nitroindazole (25 mg/kg) or carboxy-PTIO (1 mg/kg), but not L-NIO (20 mg/kg) or 7-nitroindazole (25 mg/kg). On the other hand, NO production after ischemia was significantly reduced by 7-nitroindazole (25 mg/kg), L-NIO (20 mg/kg)+7-nitroindazole (25 mg/kg) or carboxy-PTIO (1 mg/kg), but not L-NIO (20 mg/kg). The peak latency of NO production during ischemia always preceded the onset latency of anoxic depolarization in both the saline group and the carboxy-PTIO group. In the carboxy-PTIO group, the onset latency of anoxic depolarization was significantly longer than that in the saline group. Moreover, carboxy-PTIO significantly reduced the anoxic depolarization amplitude, compared with that of the saline group. These results suggest that both NOS-dependent and-independent NO formation contributes to early and sharp NO production during ischemia, and that this NO increase is, at least in part, related to the triggering of anoxic depolarization.

To clarify whether nitric oxide (NO) modifies high K(+)-evoked gamma-aminobutyric acid (GABA) release, we examined the effects of sodium nitroprusside, an NO donor; diethyldithiocarbamate, an NO trapper; dithiothreitol, a superoxide radical scavenger; and 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one, a specific guanylyl cyclase inhibitor, on high (100 mM) K(+)-evoked GABA release from rat hippocampus in vivo using microdialysis. Perfusion with 0.5 or 5 mM sodium nitroprusside significantly reduced high K(+)-evoked GABA release. Co-perfusion with 0.5 mM sodium nitroprusside and 5 mM diethyldithiocarbamate or 0.5 mM 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one significantly enhanced high K(+)-evoked GABA release. Co-perfusion with 0.5 mM sodium nitroprusside and 1 mM dithiothreitol tended to increase it. These results demonstrate that sodium nitroprusside reduces high K(+)-evoked GABA release both via an NO/cyclic GMP-dependent pathway and via an NO-dependent, but cyclic GMP-independent, pathway in rat hippocampus in vivo.

The present study investigated the effects of (-)-epigallocatechin gallate (EGCG), which is the major component of polyphenol in green tea, on nitric oxide (NO) stress-induced neuronal damage, by monitoring NO mobilizations in the intact rat hippocampus and assaying the viability of cultured rat hippocampal neurons. A 10-min ischemia increased NO (NO(3)(-)/NO(2)(-)) concentrations in the intact rat hippocampus, while EGCG (50 mg/kg i.p.) inhibited the increase by 77% without affecting hippocampal blood flow. The NO donor, sodium nitroprusside (SNP; 50 microM), produced NO (NO(3)(-)/NO(2)(-)), while EGCG inhibited it in a dose-dependent manner at concentrations ranging from 50 to 200 microM. Treatment with SNP (100 microM) reduced the viability of cultured rat hippocampal neurons to 22% of control levels, while EGCG caused it to recover to 51% for 10 microM, 73% for 20 microM, and 70% for 50 microM. Taken together, it appears that EGCG could protect against ischemic neuronal damage by deoxidizing peroxynitrate/peroxynitrite, which is converted to NO radical or hydroxy radical., (Copyright 2002 Elsevier Science B.V.)

In the present study, we examined the effects of nitric oxide (NO)-related compounds, i.e. sodium nitroprusside (NO donor), diethyldithiocarbamate (NO trapper) and dithiothreitol (superoxide radical scavenger) on release of aspartate and adenosine from rat hippocampus using electrophysiological and microdialysis methods. Perfusion with 0.05 or 0.5 mM sodium nitroprusside significantly reduced high K(+)-evoked release of aspartate during high K(+) perfusion. Perfusion with 0.5 mM sodium nitroprusside always induced seizures and significantly increased release of aspartate and adenosine during washout of sodium nitroprusside. Diethyldithiocarbamate (5 mM) reversed the effects of sodium nitroprusside. Dithiothreitol (1 mM) significantly reduced the increase in adenosine release by sodium nitroprusside. These findings indicate that adenosine release is closely related to development of seizures, which are triggered by an increase in both NO itself and in part peroxynitrite, which results in reaction with superoxide radicals.

We have recently reported that the nitric oxide (NO) donor, sodium nitroprusside (SNP), induces seizures which are associated with an increase in the basal release of aspartate and glutamate from rat hippocampus (Kaku et al., 1998). In order to determine whether taurine release occurs with SNP-induced seizures, we examined the effects of NO-related compounds, i.e., the NO trapper, diethyldithiocarbamate (DETC), the superoxide radical scavenger, dithiothreitol (DTT), the xanthine oxidase inhibitor, oxypurinol and the guanylyl cyclase inhibitor, 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ), on SNP-induced seizures and in vivo taurine release from rat hippocampus using microdialysis. Perfusion with 0.5mM SNP provoked seizures and significantly increased taurine release, with the increase in release occurring primarily during reperfusion with artificial cerebrospinal fluid lacking SNP. Perfusion with 5mM DETC significantly abolished the SNP-induced seizures and reduced taurine release during and after perfusion with the drugs. Perfusion with 1mM DTT significantly reduced both the frequency of the SNP-induced seizures and taurine release during and after perfusion with the drugs. Perfusion with 1 mM oxypurinol or 0.5 mM ODQ did not reduce the frequency of the SNP-induced seizures, but tended to decrease taurine release during and after perfusion with the drugs. These results demonstrate that SNP-induced seizures are triggered by an increase in both NO and peroxynitrite and are related to an increase in taurine release from rat hippocampus.

We investigated the effects of 7-nitroindazole, a specific inhibitor of neuronal nitric oxide (NO) synthase, on NO concentration and on blood flow in rat hippocampus after transient forebrain ischemia which was induced by 4-vessel occlusion for 10 min. NO concentration was measured directly by an NO-selective electrode method. Hippocampal blood flow was also estimated by laser Doppler flowmetry. 7-Nitroindazole [0 (vehicle), 12.5, 25, 50 or 100 mg/kg] was administered intraperitoneally 20 min before ischemia. 7-Nitroindazole at any dose used did not affect basal NO levels before ischemia. 7-Nitroindazole (25, 50 and 100 mg/kg) reduced the NO concentration significantly during post-ischemic early reperfusion. Before 10 min of ischemia and during post-ischemic early reperfusion, there were no significant differences in hippocampal basal blood flow and reactive hyperemia between vehicle- and 7-nitroindazole-treated groups. These results demonstrate that the neuronal NO synthase inhibitor, 7-nitroindazole, can effectively inhibit NO synthesis in rat hippocampus during post-ischemic early reperfusion.