小鼠子宫内胚胎电转技术方法的改良

Optimization of mouse in utero electroporation technique

目的优化小鼠子宫内胚胎电转技术,用于检测胎鼠大脑新生神经元的迁移和分化特征。方法通过选择异氟烷气体麻醉方式,改进毛细玻璃针的制备方法,减少手术中胚胎的暴露时间,优化电转参数等,对小鼠子宫内胚胎电转技术进行一系列优化。采用优化的子宫内胚胎电转技术给6只怀孕16.5 d的孕鼠进行了手术,共电转胚胎42只。结果电转后孕鼠和胚胎的存活率达到100%,其中大脑表达外源基因的胎鼠占85%±3%。结论优化的小鼠子宫内胚胎电转技术可以获得令人满意的胎鼠存活率和基因电转效率。

Objective To optimize the in utero eleetroporation protocol for transfering genes into the embryonic murine brain. Methods Critical steps of in utero electroporation were optimized, including choosing isoflurane anesthesia, improving the preparation of micropipettes, reducing the exposure time of embryos during the surgery as well as optimizing electroporation parameters. Forty-two mouse embryos from 6 mothers were subjected to in utero electroporation at gestational day 16.5 using the optimized protocol. Results The survival rates of mothers and embryos were 100%. The tranfcred gene was expressed in 85% ± 3% of electroporated embryonic brains. Conclusion Using the optimized in utero electroporation protocol, a satisfying survival rate of embryos and transfection efficiency can be achieved.