dizocilpine-maleate and linsidomine

Nitric oxide (NO) modulates the uptake and/or release of neurotransmitters through a variety of cellular mechanisms. However, the pharmacological and biochemical processes underlying these neurochemical effects of NO often remain unclear. In our study, we used immunocytochemical methods to study the effects of NO, cyclic guanosine monophosphate (cGMP), and peroxynitrite on the uptake and release of gamma-aminobutyric acid (GABA) and glycine in the turtle retina. In addition, we examined the involvement of glutamate receptors, calcium, and the GABA transporter in this GABA uptake and release. We also tested for interactions between the GABAergic and glycinergic systems. In general, we show that NO stimulated GABA release and inhibited glycine release. The NO-stimulated GABA release involved calcium-dependent or calcium-independent synaptic release or reversal of the GABA transporter. Some effects of NO on GABA release involved glutamate, cGMP, or peroxynitrite. NO promoted glycine uptake and inhibited its release, and this inhibition of glycine release was influenced by GABAergic modulation. These findings indicate that NO modulates the levels of the inhibitory transmitters GABA and glycine through several specific biochemical mechanisms in different retinal cell types and layers. Thus it appears that some of the previously described reciprocal interactions between GABA and glycine in the retina function through specific NO signaling pathways.

Topics: 6-Cyano-7-nitroquinoxaline-2,3-dione; Animals; Bicuculline; Cadmium; Citrulline; Cyclic GMP; DEET; Dizocilpine Maleate; Drug Interactions; Enzyme Inhibitors; Excitatory Amino Acid Antagonists; Free Radical Scavengers; GABA Antagonists; gamma-Aminobutyric Acid; Glycine; Immunohistochemistry; In Vitro Techniques; Molsidomine; Neural Inhibition; Nipecotic Acids; Nitric Oxide; Potassium; Retina; Silver Staining; Thiourea; Turtles; Vigabatrin

Free radicals are suggested to play an important role in hypoxic-ischemic neuronal death. However, the importance in human disease is not known. Furthermore, whether posthypoxic free radical formation mainly occurs in endothelium and neutrophils, or whether neuronal production is important, is not finally determined. To study this we differentiated human Ntera2 teratocarcinoma cells into postmitotic NT2-N neurons and exposed them to free radicals, hypoxia, or oxygen and glucose deprivation. These cells are devoid of nitric oxide synthase, and we hypothesized that free radicals are important mediators downstream of N-methyl-D-aspartate stimulation. Production of free radicals, evaluated with the fluorescent dyes dihydrorhodamine and 2',7'-dichlorodihydrofluorescein, was significantly higher in neurons deprived of oxygen and glucose after 40 min of reoxygenation than in normoxic cells. The antioxidant trolox, the flavonoid quercetin, thiopental, and the N-methyl-D-aspartate-glutamate receptor antagonist MK-801 reduced the formation of free radicals. Treatment with the flavonoid rutin (86 +/- 16% of hypoxic cells without drug, p < 0.01), trolox (86 +/- 20%, p < 0.01), and MK-801 (57 +/- 12%, p < 0.01) reduced lactate dehydrogenase release after 6 h of hypoxia. Trolox, salicylate, and quercetin also significantly reduced lactate dehydrogenase release after 3 h of oxygen and glucose deprivation. The protection offered by these antioxidants was, however, limited compared with the effect of MK-801. We conclude that oxygen and glucose deprivation causes a moderate increase in the formation of free radicals in NT2-N neurons that can be inhibited by antioxidants and by blocking of the N-methyl-D-aspartate-glutamate receptor. Although MK-801 conveys profound protection, antioxidants provide only a limited improvement in neuronal survival. Thus in this model, mechanisms downstream of the N-methyl-D-aspartate-glutamate receptor other than free radicals and nitric oxide have to be invoked.

Topics: Antioxidants; Cell Death; Cell Hypoxia; Dizocilpine Maleate; Enzyme Inhibitors; Excitatory Amino Acid Antagonists; Fluorescent Dyes; Free Radicals; Glucose; Humans; L-Lactate Dehydrogenase; Molsidomine; Neurons; Neuroprotective Agents; Oxygen; Reactive Oxygen Species; Tumor Cells, Cultured

The purpose of this study was to characterize behavioral interactions between nitric oxide synthase (NOS) inhibitors and serotonergic 5-HT2 receptors. In the present study, NOS inhibitors, N(G)-nitro-L-arginine, N(G)-nitro-L-arginine methylester, N(G)-monomethyl-L-arginine, 7-nitroindazole, trifluoperazine and NO scavenger, methylene blue markedly enhanced 5-hydroxytryptamine (5-HT)-induced selective serotonergic behavior, the head twitch response (HTR), in mice. However NO generators, sodium nitroprusside, 3-morpholinosydnonimine and S-nitroso-N-acetylpenicillamine as well as NO precursor, L-arginine markedly inhibited 5-HT induced HTR in mice. In the previous study, it was demonstrated that the N-methyl-D-aspartate (NMDA) receptor antagonists markedly enhanced 5-HT-induced selective serotonergic behavior, HTR, whereas NMDA itself inhibited 5-HT-induced HTR in mice. In the present study, it was demonstrated that the inhibition by a NMDA receptor agonist, NMDA of 5-HT-induced HTR was reversed by the treatment with NOS inhibitors, N(G)-nitro-L-arginine and N(G)-nitro-L-arginine methylester. The suppressive action by a NO generator, S-nitroso-N-acetylpenicillamine of 5-HT-induced HTR was also reversed by the treatment with NMDA receptor antagonists, MK-801 and dextromethorphan. These results have shown that the NO system is located down stream of NMDA receptors involved in modulation of 5-HT2-mediated HTR. Therefore, the enhanced effects of NOS inhibitors on 5-HT-induced HTR support experimental evidence for the NO/5-HT2 as well as NMDA/5-HT2 receptor interactions indicating that NO plays an important role in the glutamatergic modulation of the serotonergic function at the 5-HT2 receptor.

Topics: Animals; Arginine; Dextromethorphan; Dizocilpine Maleate; Dose-Response Relationship, Drug; Enzyme Inhibitors; Head Movements; Male; Methylene Blue; Mice; Mice, Inbred ICR; Molsidomine; N-Methylaspartate; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase; Nitroprusside; Penicillamine; Receptors, Serotonin; Serotonin; Serotonin Antagonists

Topics: Amino Acid Oxidoreductases; Analysis of Variance; Animals; Arginine; Cerebral Infarction; Dizocilpine Maleate; Ischemic Attack, Transient; Male; Mice; Molsidomine; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Vasodilator Agents