帕金森病基因α-Synuclein过表达对秀丽隐杆线虫毒性作用的研究

Toxic effect of Parkinson's disease gene α-Synuclein on caenorhabditis elegans

目的:利用载体pPD95.86建立携带帕金森病基因α-Synuclein(α-SYN)的秀丽隐杆线虫(秀丽线虫)模型,探讨α-SYN过表达对秀丽线虫的毒性作用。方法:构建同时表达α-SYN和绿色荧光蛋白(GFP)的重组质粒pPD95.86::α-SYN,采用野生型N2秀丽线虫,通过显微注射和基因整合筛选获得pPD95.86转基因线虫和pPD95.86::α-SYN转基因线虫。同步化野生型N2秀丽线虫和两种转基因线虫,以pPD95.86::α-SYN转基因线虫为实验组,以pPD95.86转基因线虫为条件对照组,以N2野生型线虫为空白对照组,观察3组线虫的寿命、运动能力和热休克条件下的存活情况。结果:与两对照组比较,pPD95.86::α-SYN转基因线虫的寿命明显缩短,运动能力显著下降,热休克条件下其存活能力减弱(P均<0.01)。结论:帕金森病基因α-SYN的过表达对秀丽线虫具有明显的毒性作用,并且该毒性在热休克条件下加重。

Objective： To establish a transgenic caenorhabditis elegans （C. elegans） model of human ct-synu- clein with the vector pPD95.86 and explore the toxic effects of α-synuclein overexpression on the life-span of elegans. Methods： Human α-synuclein and the green fluorescent protein （GFP） were fused and cloned into the vector pPD95.86. Transgenic C. elegans with pPD95.86 and pPD95.86： ： α-SYN were produced by microinjecting the recombinant plasmid into wild-type worms and gene integration. Transgenic C. elegans with pPD95.86 ： α-SYN were divided into experimental group, transgenic C. elegans with pPD95.86 into control conditions, and N2 wild-type C. elegans into blank control group The effects of α-synuclein expression on the life-span were studied. Results： Compared with two control group, transgenic C. elegans with pPD95.86： α-SYN had significant shorter life span and their motor function was impaired （ all P 〈 0.01 ）. Exposure to the heat stress significantly compromised the viability of transgenic C. elegans expressing human α-synuclein （P 〈0. 01 ）. Conclusion： Human α-synuclein expressing in muscle cells of C. elegans decreases the life-span of elegans