In utero intraventricular injection and electroporation of E16 rat embryos.
In utero intraventricular injection and electroporation of E16 rat embryos.
复制标题
DOI：
10.3791/236
复制
发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
Kriegstein, Arnold
中科院分区：
文献类型：
--
作者：
Walantus, William;Elias, Laura;Kriegstein, Arnold
胚胎大鼠新皮质的宫内注射和电穿孔为操纵侧脑室壁上的单个祖细胞提供了一个强大的工具。这项技术现在被广泛应用于通过过度表达或敲除基因并观察对细胞增殖、迁移和分化的影响来研究涉及皮质生成的过程。与传统的基因敲除策略相比，宫内电穿孔提供了一种在特定的时间窗口内操纵细胞群体的快速手段。在这一视频协议中，我们概述了为手术准备大鼠的实验方法，通过卵巢手术暴露子宫角，将DNA注入发育中的胚胎的侧脑室，将DNA电穿孔到排列在侧壁上的祖细胞中，以及在手术后照顾动物。我们的实验室在E15-E21大鼠的手术中使用了这种方案，但如本视频所示，最常见的是在E16进行的手术。
In-utero in-vivo injection and electroporation of the embryonic rat neocortex provides a powerful tool for the manipulation of individual progenitors lining the walls of the lateral ventricle. This technique is now widely used to study the processes involved in corticogenesis by over-expressing or knocking down genes and observing the effects on cellular proliferation, migration, and differentiation. In comparison to traditional knockout strategies, in-utero electroporation provides a rapid means to manipulate a population of cells during a specific temporal window. In this video protocol we outline the experimental methodology for preparing rats for surgery, exposing the uterine horns through laporatomy, injecting DNA into the lateral ventricles of the developing embryo, electroporating DNA into the progenitors lining the lateral wall, and caring for animals post-surgery. Our laboratory uses this protocol for surgeries on E15-E21 rats, however it is most commonly performed at E16 as shown in this video.