Neuronal death in cultured murine cortical cells is induced by inhibition of GAPDH and triosephosphate isomerase.
Polyglutamine-containing proteins expressed in the CAG repeat diseases Huntington's disease and dentatorubralpallidoluyisian atrophy have recently been suggested to inhibit the key glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). To examine the consequences of GAPDH inhibition upon neuronal survival, we exposed murine neocortical cell cultures to the inhibitor of GAPDH and triosephosphate isomerase, alpha-monochlorohydrin. Cultures exposed to 6-15 mM alpha-monochlorohydrin for 48 h exhibited an increase in dihydroxyacetone phosphate and a decrease in neuronal ATP that was followed by progressive neuronal death; some glial death occurred at high drug concentrations. The neuronal death was characterized by cell body shrinkage and chromatin condensation and was sensitive to cycloheximide and to the caspase inhibitors Z-Val-Ala-Asp fluoromethylketone and tert-butoxycarbonyl-Asp fluoromethylketone. Neurons in striatal cell cultures were more vulnerable to death induced by exposure to alpha-monochlorohydrin, except that NADPH-diaphorase(+) neurons were selectively spared. Repeated addition of the glycolytic endpoint metabolite pyruvate to the bathing medium attenuated both the drop in neuronal ATP and the neuronal cell death.
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
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D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000201:enzymology / Q000473:pathology / Q000502:physiology*; D004099:Dihydroxyacetone Phosphate / Q000037:antagonists & inhibitors; D004351:Drug Resistance; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors*; D051379:Mice; D009252:NADPH Dehydrogenase / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D019289:Pyruvic Acid / Q000494:pharmacology; D014305:Triose-Phosphate Isomerase / Q000037:antagonists & inhibitors*; D000517:alpha-Chlorohydrin / Q000494:pharmacology
D004099:Dihydroxyacetone Phosphate; D019289:Pyruvic Acid; D000255:Adenosine Triphosphate; D000517:alpha-Chlorohydrin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D009252:NADPH Dehydrogenase; D014305:Triose-Phosphate Isomerase