Gangliosides attenuate ethanol-induced apoptosis in rat cerebellar granule neurons.
Ethanol significantly enhances cell death of differentiated rat cerebellar granule neurons on culture in a serum-free medium containing a depolarizing concentration of KCl (25 mM), 5 microM MK-801 (an NMDA receptor antagonist), and 20-200 mM ethanol for 1-4 days. Cell death augmented by ethanol was concentration- and time-dependent with neurons displaying hallmark apoptotic morphology and DNA fragmentation that correlated with the activation of cytosolic caspase-3. Inclusion of 5 microM MK-801 or 100 microM glycine in culture media did not alter rates of cell death indicating ethanol toxicity is mediated via an NMDA receptor-independent pathway. Preincubation with 50 microM gangliosides GM1, GD1a, GD1b or GT1b for 2 h, or preincubation with 10 microM LIGA20 (a semisynthetic GM1 with N-dichloroacetylsphingosine) for 10 min, attenuated caspase-3 activity and ethanol-induced cell death. Data show native gangliosides and a synthetic derivative are potently neuroprotective in this model of ethanol toxicity, and potentially serve as useful probes to further unravel the mechanisms relevant to neuronal apoptosis.
Saito
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Department of Neurochemistry, The Nathan S. Kline Institute for Psychiatric Research and New York University, Orangeburg 10962, USA. Marsaito@NKI.rfmh.org
Saito
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Berg
M J
MJ
Guidotti
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Marks
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D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects*; D003594:Cytoplasmic Granules / Q000187:drug effects; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004789:Enzyme Activation; D000431:Ethanol / Q000633:toxicity*; D005677:G(M1) Ganglioside / Q000031:analogs & derivatives / Q000494:pharmacology; D005732:Gangliosides / Q000494:pharmacology*; D005998:Glycine / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013110:Sphingosine / Q000031:analogs & derivatives / Q000494:pharmacology
D005732:Gangliosides; C086919:LIGA 20; D005677:G(M1) Ganglioside; D000431:Ethanol; D016291:Dizocilpine Maleate; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D013110:Sphingosine; D005998:Glycine