Establishing a novel C. elegans model to investigate the role of autophagy in amyotrophic lateral sclerosis.
Establishing a novel C. elegans model to investigate the role of autophagy in amyotrophic lateral sclerosis.
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建立新型线虫模型以研究自噬在肌萎缩侧索硬化症中的作用
DOI：
10.1038/aps.2012.190
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发表时间：
2013-05
影响因子：
8.2
通讯作者：
Le, Wei-dong
中科院分区：
文献类型：
--
作者：
Li, Jia;Huang, Kai-xing;Le, Wei-dong
目的：建立肌萎缩侧索硬化症（ALS）的秀丽隐杆线虫模型，并评估自噬在该疾病中的作用。方法：产生在 GABA 能运动神经元中表达铜、锌超氧化物歧化酶（SOD1）G93A 突变体的稳定转基因线虫。用荧光显微镜观察运动神经元中的轴突引导和蛋白质聚集。进行麻痹测定以评估转基因线虫的运动功能。使用实时 PCR 分析 daf-2 (e1370) 突变体中自噬基因的表达。使用报告基因GFP::LGG-1验证运动神经元中是否诱导自噬。结果：运动神经元中G93A SOD1的表达引起年龄依赖性运动缺陷，并伴有显着的SOD1聚集和轴突引导失败。 12 天后，超过 80% 的 G93A 线虫变得麻痹，而不到 10% 的对照线虫表现出麻痹表型。在秀丽隐杆线虫的daf-2（e1370）突变体中，自噬基因bec-1、atg-7、lgg-1和atg-18的水平上调约1.5倍，unc-51的水平增加约4倍，运动神经元中的自噬体显着增加。将 daf-2 (e1370) 突变与 G93A SOD1 突变蠕虫杂交可显着改善运动缺陷、SOD1 聚集和轴突引导失败。 结论：线虫运动神经元中 G93A SOD1 的表达导致 ALS 的特征性改变。自噬的增加可以保护秀丽隐杆线虫运动神经元免受突变体 SOD1 的毒性。
Aim:To develop a C. elegans model of amyotrophic lateral sclerosis (ALS) and to evaluate the role of autophagy in the disease.Methods:Stable transgenic worms expressing the G93A mutant form of Cu, Zn-superoxide dismutase (SOD1) in GABAergic motor neurons were generated. Axon guidance and protein aggregation in the motor neurons were observed with fluorescence microscopy. A paralysis assay was performed to evaluate the motor function of the transgenic worms. The expression of autophagic genes in daf-2 (e1370) mutants was analyzed using real-time PCR. The reporter GFP:: LGG-1 was used to verify whether autophagy was induced in motor neurons.Results:Expression of G93A SOD1 in motor neurons caused age-dependent motor defects accompanied by significant SOD1 aggregation and axon guidance failure. After 12 d, over 80% of the G93A worms became paralyzed, whereas less than 10% of the controls showed a paralytic phenotype. In the daf-2 (e1370) mutants of C. elegans, the levels of autophagic genes bec-1, atg-7, lgg-1, and atg-18 were upregulated by approximately 1.5-fold, the level of unc-51 increased by approximately fourfold, and autophagosomes in motor neurons was markedly increased. Crossing the daf-2 (e1370) mutation into the G93A SOD1 mutant worms significantly ameliorated the motor defects, SOD1 aggregation, and axon guidance failure.Conclusion:G93A SOD1 expression in motor neurons of C. elegans results in characteristic alterations of ALS. Increased autophagy protects C. elegans motor neurons against the toxicity of mutant SOD1.
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影响因子：
4.8
作者：
Kabuta, Tomohiro;Suzuki, Yasuyuki;Wada, Keiji
通讯作者：
Wada, Keiji
影响因子：
13.3
作者：
Li, Liang;Zhang, Xiaojie;Le, Weidong
通讯作者：
Le, Weidong
影响因子：
14.8
作者：
Sarkar, Sovan;Perlstein, Ethan O.;Imarisio, Sara;Pineau, Sandra;Cordenier, Axelle;Maglathlin, Rebecca L.;Webster, John A.;Lewis, Timothy A.;O'Kane, Cahir J.;Schreiber, Stuart L.;Rubinsztein, David C.
通讯作者：
Rubinsztein, David C.
影响因子：
13.3
作者：
Florez-McClure, Maria L.;Hohsfield, Lindsay A.;Link, Christopher D.
通讯作者：
Link, Christopher D.
影响因子：
2.9
作者：
Morimoto, Nobutoshi;Nagai, Makiko;Abe, Koji
通讯作者：
Abe, Koji