Acute NMDA toxicity in cultured rat cerebellar granule neurons is accompanied by autophagy induction and late onset autophagic cell death phenotype.
Acute NMDA toxicity in cultured rat cerebellar granule neurons is accompanied by autophagy induction and late onset autophagic cell death phenotype.
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DOI：
10.1186/1471-2202-11-21
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发表时间：
2010-02-18
期刊：
影响因子：
2.4
通讯作者：
Wang KK
中科院分区：
文献类型：
--
作者：
Sadasivan S;Zhang Z;Larner SF;Liu MC;Zheng W;Kobeissy FH;Hayes RL;Wang KK
自噬是一种细胞内对应激的反应，其特征是称为自噬体的双层膜胞质囊泡。已知延长自噬会导致自噬（II 型）细胞死亡。本研究探讨了在受到兴奋性毒素 N-甲基-D-天冬氨酸 (NMDA) 攻击的培养的小脑颗粒神经元中自噬反应的潜在作用。 NMDA 暴露早在治疗后 3 小时就在细胞体和神经突中诱导轻链 3 (LC-3) 免疫阳性和单丹磺酰尸胺 (MDC) 荧光染料标记的自噬体形成。在接触 NMDA 后，还观察到 Beclin-1 和自噬体靶向 LC3-II 水平升高。培养物长时间暴露于 NMDA（8-24 小时）会产生 MDC、LC3 阳性自噬体，同时伴随着 NMDA 攻击的神经退行性阶段。溶酶体抑制研究还表明，NMDA 治疗使自噬体相关的 LC3-II 从正常的溶酶体降解途径转移。自噬抑制剂 3-甲基腺嘌呤显着降低 NMDA 诱导的 LC3-II/LC3-I 比值增加和自噬体的积累，并抑制 NMDA 介导的神经元死亡。 ATG7 siRNA 研究还显示 NMDA 治疗后的神经保护作用。总的来说，这项研究表明，在长期暴露于兴奋毒素的培养神经元中，自噬机制被强烈诱导，而溶酶体途径的自噬体清除可能会受到损害。我们的数据进一步表明，延长的自噬导致 NMDA 介导的兴奋性毒性中的细胞死亡。
Autophagy, an intracellular response to stress, is characterized by double membrane cytosolic vesicles called autophagosomes. Prolonged autophagy is known to result in autophagic (Type II) cell death. This study examined the potential role of an autophagic response in cultured cerebellar granule neurons challenged with excitotoxin N-methyl-D-aspartate (NMDA). NMDA exposure induced light chain-3 (LC-3)-immunopositive and monodansylcadaverine (MDC) fluorescent dye-labeled autophagosome formation in both cell bodies and neurites as early as 3 hours post-treatment. Elevated levels of Beclin-1 and the autophagosome-targeting LC3-II were also observed following NMDA exposure. Prolonged exposure of the cultures to NMDA (8-24 h) generated MDC-, LC3-positive autophagosomal bodies, concomitant with the neurodegenerative phase of NMDA challenge. Lysosomal inhibition studies also suggest that NMDA-treatment diverted the autophagosome-associated LC3-II from the normal lysosomal degradation pathway. Autophagy inhibitor 3-methyladenine significantly reduced NMDA-induced LC3-II/LC3-I ratio increase, accumulation of autophagosomes, and suppressed NMDA-mediated neuronal death. ATG7 siRNA studies also showed neuroprotective effects following NMDA treatment. Collectively, this study shows that autophagy machinery is robustly induced in cultured neurons subjected to prolonged exposure to excitotoxin, while autophagosome clearance by lysosomal pathway might be impaired. Our data further show that prolonged autophagy contributes to cell death in NMDA-mediated excitotoxicity.
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影响因子：
16.2
作者：
ANKARCRONA, M;DYPBUKT, JM;NICOTERA, P
通讯作者：
NICOTERA, P
影响因子：
3
作者：
KOH, JY;CHOI, DW
通讯作者：
CHOI, DW
影响因子：
6.1
作者：
Carloni, Silvia;Buonocore, Giuseppe;Balduini, Walter
通讯作者：
Balduini, Walter
影响因子：
10.6
作者：
Boland, Barry;Nixon, Ralph A.
通讯作者：
Nixon, Ralph A.
影响因子：
4
作者：
González-Polo, RA;Boya, P;Kroemer, G
通讯作者：
Kroemer, G