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.
Sheline
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CT
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Choi
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DW
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