Glutamate receptors induce a burst of superoxide via activation of nitric oxide synthase in arginine-depleted neurons.
We have previously shown in cultured cerebellar granule neurons (Lafon-Cazal, M., Pietri, S., Culcasi, M., and Bockaert, J. (1993) Nature 364, 535-537) that upon N-methyl-D-aspartate stimulation, a nitric oxide synthase (NOS)-independent, arachidonic acid-dependent generation of superoxide free radicals (O2-.) is observed after a lag time of 10-15 min. Using the electron spin resonance spin trapping technique, we show that N-methyl-D-aspartate stimulation produced a more rapid burst of O2-. in L-arginine (L-Arg)-depleted neurons. These O2-. radicals are synthesized by NOS. KCl and kainate, which also stimulated NOS in these neurons, produced this rapid burst of O2-., which was blocked as follows: (a) in the presence of L-NG-nitro-arginine (L-Narg), and (b) by L-Arg repletion. This burst of O2-. was arachidonic acid-independent, and its time course was similar to that of nitric oxide production. It was also responsible for a weak but significant cell death that was suppressed by L-Narg and L-Arg.
Culcasi
M
M
Structure et Réactivité des Espaces Paramagnétiques-Centre National de la Recherche Scientifique Unité de Recherche Associée 1412, Université de Provence, Marseille, France.
Lafon-Cazal
M
M
Pietri
S
S
Bockaert
J
J
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001119:Arginase / Q000378:metabolism; D001120:Arginine / Q000172:deficiency / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016923:Cell Death; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D004789:Enzyme Activation; D008297:Male; D051379:Mice; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000378:metabolism*; D019001:Nitric Oxide Synthase; D017470:Receptors, Glutamate / Q000502:physiology*; D013481:Superoxides / Q000378:metabolism*
D017470:Receptors, Glutamate; D013481:Superoxides; D016202:N-Methylaspartate; D001120:Arginine; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D001119:Arginase; D006152:Cyclic GMP