Dual effect of glycine on NMDA-induced neurotoxicity in rat cortical cultures.
To examine the roles of glycine in neurotoxicity caused by NMDA, primary rat cortical cultures were exposed to 100-300 microM NMDA plus glycine (0-3000 microM) or other glycine analogs in a simple saline solution, and toxicity was assessed by the amount of lactate dehydrogenase (LDH) released from the cultures. NMDA-induced neurotoxicity was abolished by 100 microM D-2-amino-5-phosphonovaleric acid (D-APV), phencyclidine (IC50, 4.1 microM), and Mg (IC50, 7.5 mM), or by reducing [Ca]0 to 0.1 mM. NMDA-induced neurotoxicity could also be abolished by 7-chlorokynurenic acid (IC50, 8.6 microM), suggesting the presence of residual glycine in the culture medium (confirmed by high-performance liquid chromatography measurement). Moreover, in the presence of 30 microM 7-chlorokynurenic acid, glycine, D-serine, D-alanine, beta-fluoro-D-alanine, and 1-aminocyclopropanecarboxylic acid could restore the neurotoxic action of NMDA, and their relative potencies and relative efficacies were the same as measured in electrophysiological assays in Xenopus oocytes or cultured neurons. The addition of greater than 100 microM glycine doubled the excitotoxic effect of NMDA. The potency of glycine was low (EC50, 27 microM), and this effect was not due to a direct action on the NMDA receptor. The above-mentioned agonists were unable to substitute for glycine, even at high concentrations (1 mM). On the other hand, beta-alanine, taurine, and GABA (1 mM) did potentiate NMDA neurotoxicity, and strychnine (IC50, 550 nM) could greatly reduce neurotoxicity in the presence of 1 mM glycine plus 300 microM NMDA.(ABSTRACT TRUNCATED AT 250 WORDS)
McNamara
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Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill 27599.
Dingledine
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D015763:2-Amino-5-phosphonovalerate / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D004357:Drug Synergism; D005998:Glycine / Q000494:pharmacology*; D007736:Kynurenic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016202:N-Methylaspartate / Q000494:pharmacology*; D009410:Nerve Degeneration; D009474:Neurons / Q000187:drug effects / Q000201:enzymology; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D010622:Phencyclidine / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D018009:Receptors, Glycine; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000502:physiology; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000502:physiology; D013654:Taurine / Q000494:pharmacology; D014981:Xenopus; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D018009:Receptors, Glycine; D016194:Receptors, N-Methyl-D-Aspartate; D017981:Receptors, Neurotransmitter; D013654:Taurine; D005680:gamma-Aminobutyric Acid; D016202:N-Methylaspartate; D015763:2-Amino-5-phosphonovalerate; D007770:L-Lactate Dehydrogenase; D007736:Kynurenic Acid; D010622:Phencyclidine; C057013:7-chlorokynurenic acid; D005998:Glycine