Sheline C T, Choi D W
Department of Neurology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Neurobiol Dis. 1998 Jul;5(1):47-54. doi: 10.1006/nbdi.1998.0177.
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.
最近有研究表明，在CAG重复序列疾病亨廷顿舞蹈病和齿状核红核苍白球路易体萎缩症中表达的含多聚谷氨酰胺蛋白会抑制关键糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH）。为了研究GAPDH抑制对神经元存活的影响，我们将小鼠新皮质细胞培养物暴露于GAPDH和磷酸丙糖异构酶的抑制剂α-一氯丙醇中。暴露于6-15 mM α-一氯丙醇48小时的培养物中，磷酸二羟丙酮增加，神经元ATP减少，随后神经元逐渐死亡；在高药物浓度下会发生一些胶质细胞死亡。神经元死亡的特征是细胞体收缩和染色质浓缩，并且对环己酰亚胺以及半胱天冬酶抑制剂Z-缬氨酸-丙氨酸-天冬氨酸氟甲基酮和叔丁氧羰基-天冬氨酸氟甲基酮敏感。纹状体细胞培养物中的神经元更容易受到暴露于α-一氯丙醇诱导的死亡影响，不过NADPH-黄递酶（+）神经元被选择性地保留下来。向培养液中反复添加糖酵解终产物丙酮酸可减轻神经元ATP的下降和神经元细胞死亡。
