摘要
【目的】探讨肌萎缩侧索硬化症基因TDP43致病突变对斑马鱼毒性的影响。【方法】以野生型AB系斑马鱼作为研究对象,构建携带红色荧光蛋白(RFP)的TDP43基因野生型(WT)和突变型(S292N)质粒,通过显微注射法转染斑马鱼单细胞期胚胎,建立转基因斑马鱼模型。通过比较正常AB系斑马鱼、TDP43-WT转基因斑马鱼以及TDP43-S292N基因突变斑马鱼的胚胎发育、运动能力等,探讨TDP43基因突变的毒性作用。【结果】通过显微注射,成功获得表达红色荧光的TDP43-WT及TDP43-S292N转基因鱼。与正常AB系斑马鱼及TDP43-WT转基因斑马鱼相比,TDP43-S292N基因突变斑马鱼的胚胎死亡率及畸形率升高,胚胎孵化率、胚胎摆尾次数以及孵化后运动活力降低(P<0.01)。【结论】TDP43基因突变影响斑马鱼胚胎发育并降低孵化后的运动功能。
[ Objective ] To investigate the toxicity of amyotrophic lateral sclerosis gene TDP43 mutations on zebrafish. [ Methods ] TDP43 Wild Type and S292N mutation were successfully cloned into the red fluorescent protein tagging vectors. The plasmids were then microinjected into zebrafish embryos to obtain the transgenic lines. The toxicity was evaluated by assessing the embryonic development and motor function in normal AB, TDPg3-WT, and TDP43-S292N transgenic zebrallsh. [Results] Red fluorescent was evident in both TDP43-WT and TDPg3-s292N transgenic zebrafish following microinjection. Compared with normal AB and TDP43- WT transgenic zebrafish, TDPg3-s292N mutant had higher embryonic mortality and abnormality rates and lower embryonic hatch rates. The tail beat frequency and motor ability was also significantly lower in TDPg3-s292N mutant （P 〈 0.01）. [ Conclusion] Expression of human mutant TDP43 in zebrafish disturbs the normal embryonic development and reduced the motor ability.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2014年第4期481-487,共7页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(81100936)
广东省自然科学基金(S20110110004860)
