In vivo electroporation in the embryonic mouse central nervous system
In vivo electroporation in the embryonic mouse central nervous system
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DOI：
10.1038/nprot.2006.276
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Saito, Tetsuichiro
中科院分区：
文献类型：
--
作者：
Saito, Tetsuichiro
该协议描述了胚胎小鼠神经系统中体内电穿孔的基本方法。将DNA微注射到脑室或脊髓中央管后的电脉冲传递，可以有效地将基因转染到神经系统中。转染由镊子型电极促进，该电极容纳子宫和/或包含胚胎的蛋黄囊。单个怀孕小鼠中的十个胚胎可以在30分钟内进行操作。超过90％的手术胚胎存活，其中90％以上的幸存者适当地表达了转染的基因。神经元中的基因表达持续很长一段时间，即使在产后阶段，电穿孔后也是如此。因此，该方法不仅可以用于分析基因在胚胎发育中的作用，而且还可以通过神经系统（例如学习）来分析基因的作用。
This protocol describes a basic method for in vivo electroporation in the nervous system of embryonic mice. Delivery of electric pulses following microinjection of DNA into the brain ventricle or the spinal cord central canal enables efficient transfection of genes into the nervous system. Transfection is facilitated by forceps-type electrodes, which hold the uterus and/or the yolk sac containing the embryo. More than ten embryos in a single pregnant mouse can be operated on within 30 min. More than 90% of operated embryos survive and more than 90% of these survivors express the transfected genes appropriately. Gene expression in neurons persists for a long time, even at postnatal stages, after electroporation. Thus, this method could be used to analyze roles of genes not only in embryonic development but also in higher order function of the nervous system, such as learning.