Chronic ethanol exposure potentiates NMDA excitotoxicity in cerebral cortical neurons.
The effect of acute and chronic ethanol exposure on excitotoxicity in cultured rat cerebral cortical neurons was examined. Neuronal death was quantitated by measuring the accumulation of lactate dehydrogenase (LDH) in the culture media 20 h after exposure to NMDA. Addition of NMDA (25-100 microM) to the culture dishes for 25 min in Mg(2+)-free buffer resulted in a dose-dependent increase in LDH accumulation. Phase-contrast microscopy revealed obvious signs of cellular injury as evidenced by granulation and disintegration of cell bodies and neuritic processes. Chronic exposure of neuronal cultures to ethanol (100 mM) for 96 h followed by its removal before NMDA exposure, significantly increased NMDA-stimulated LDH release by 36 and 22% in response to 25 microM and 50 microM NMDA, respectively. Neither basal LDH release nor that in response to maximal NMDA (100 microM) stimulation was altered by chronic alcohol exposure. In contrast to the effects of chronic ethanol on NMDA neurotoxicity, inclusion of ethanol (100 mM) only during the NMDA exposure period significantly reduced LDH release by approximately 50% in both control and chronically treated dishes. This reduction by acute ethanol was also observed under phase-contrast microscopy as a lack of development of granulation and a sparing of disintegration of neuritic processes. These results indicate that chronic exposure of ethanol to cerebral cortical neurons in culture can sensitize neurons to excitotoxic NMDA receptor activation.
Chandler
L J
LJ
Department of Pharmacology, University of Florida College of Medicine, Gainesville 32601-0267.
Newsom
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Sumners
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Crews
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D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002540:Cerebral Cortex / Q000166:cytology*; D004357:Drug Synergism; D000431:Ethanol / Q000008:administration & dosage / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D016202:N-Methylaspartate / Q000008:administration & dosage / Q000494:pharmacology / Q000633:toxicity*; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D009474:Neurons / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D051381:Rats
D000431:Ethanol; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase; D008274:Magnesium