Modulation of glutamate release from rat hippocampal synaptosomes by nitric oxide.
Nitric Oxide 1997 Aug;1(4):315-29 Unique Identifier : MEDLINE 98368277 Sequeira SM; Ambrosio AF; Malva JO; Carvalho AP; Carvalho CM; Center for Neuroscience of Coimbra, Department of Zoology and Faculty; of Medicine, University of Coimbra, Portugal.
Abstract:
We used hippocampal synaptosomes to study the effect of NO originating from NO donors and from the activation of the NO synthase on the Ca2+- dependent release of glutamate due to 4-aminopyridine (4-AP) depolarization. We distinguished between the effects of NO on the exocytotic and on the carrier-mediated release of glutamate, which we found to be related to an increase in cGMP content and to a reduction of the ATP/ADP ratio, respectively. The NO donor hydroxylamine, at concentrations < or = 0.3 mM, inhibited the Ca2+-dependent glutamate release evoked by 4-AP, and addition of the NO donor, NOC-7, had a similar effect, which was reversed by the NO scavenger, carboxy-PTIO. Increasing the activity of NO synthase by addition of L-arginine also led to a decrease in the Ca2+-dependent release of glutamate induced by 4-AP, and this effect was reversed by inhibiting NO synthase with NG- nitro-L-arginine. This depression of the exocytotic release of glutamate was accompanied by an increase in cGMP levels due to the stimulation of soluble guanylyl cyclase by NO, produced either by the NO donors (hydroxylamine <0.3 mM) or by the endogenous NO synthase, but no significant decrease in ATP/ADP ratio was observed. However, at concentrations > or = 0.3 mM, hydroxylamine drastically increased the basal release and completely inhibited the Ca2+-dependent release of glutamate (IC50 = 168 microM). At these higher levels of NO, cGMP levels dropped to about 40% of the maximal values obtained at lower concentrations, and the ATP/ADP ratio decreased to about 50% (at 0.3 mM hydroxylamine). The large increase in the basal release could be partially inhibited by L-trans-2,4-PDC, previously loaded into the synaptosomes, suggesting that the nonexocytotic basal release occurred by reversal of the glutamate carrier. Therefore, the increase in cGMP induced by NO stimulation of the guanylyl cyclase decreases the exocytotic release of glutamate, but higher NO levels reduce the ATP/ADP ratio by inhibiting mitochondrial function, which therefore causes the massive release of cytosolic glutamate through the glutamate carrier.
Keywords: JOURNAL ARTICLE Adenosine Diphosphate/metabolism Adenosine Triphosphate/metabolism Animal Calcium/metabolism Cyclic GMP/metabolism Glutamic Acid/*metabolism Hippocampus/*drug effects/enzymology/metabolism Hydroxylamine/pharmacology Male Nitric Oxide/chemistry/*pharmacology Nitric-Oxide Synthase/metabolism Rats Rats, Wistar Support, Non-U.S. Gov't Synaptosomes/*drug effects/enzymology/metabolism 4-Aminopyridine/pharmacology
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