Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus-and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse
Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus-and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse
复制标题
DOI：
10.3791/53303
复制
发表时间：
2016-01-01
影响因子：
1.2
通讯作者：
Baumgart, Nadine
中科院分区：
文献类型：
--
作者：
Baumgart, Jan;Baumgart, Nadine
子宫内电穿孔是一种广泛使用的技术，用于快速有效地时空操纵啮齿动物中枢神经系统中的各种基因。所需基因的过表达与shRNA介导的功能丧失研究一样可能。因此，它提供了广泛的应用。靶向不同区域中的特定细胞的可行性进一步增加了这种非常有用的方法的潜在应用范围。为了有效地靶向特定区域的知识的微妙之处，如胚胎阶段，电压施加，最重要的是电极的位置，是必不可少的。在这里，我们提供了一个详细的协议，允许特定的和有效的子宫内电穿孔的几个地区的C57 BL/6小鼠中枢神经系统。特别是，它显示了如何识别区域发展成压后皮质，运动皮质，体感皮质，梨状皮质，角amnus 1-3，齿状回，纹状体，外侧隔核，丘脑和下丘脑。对于关于适当胚胎阶段的该信息，提供用于相应胚胎阶段的适当电压。最重要的是，描绘了指示正极的适当位置的角度图。该标准化方案有助于促进有效的子宫内电穿孔，这也可能导致动物数量减少。
In utero electroporation is a widely used technique for fast and efficient spatiotemporal manipulation of various genes in the rodent central nervous system. Overexpression of desired genes is just as possible as shRNA mediated loss-of-function studies. Therefore it offers a wide range of applications. The feasibility to target particular cells in a distinct area further increases the range of potential applications of this very useful method. For efficiently targeting specific regions knowledge about the subtleties, such as the embryonic stage, the voltage to apply and most importantly the position of the electrodes, is indispensable.Here, we provide a detailed protocol that allows for specific and efficient in utero electroporation of several regions of the C57BL/6 mouse central nervous system. In particular it is shown how to transfect regions the develop into the retrosplenial cortex, the motor cortex, the somatosensory cortex, the piriform cortex, the cornu ammonis 1-3, the dentate gyrus, the striatum, the lateral septal nucleus, the thalamus and the hypothalamus. For this information about the appropriate embryonic stage, the appropriate voltage for the corresponding embryonic stage is provided. Most importantly an angle-map, which indicates the appropriate position of the positive pole, is depicted. This standardized protocol helps to facilitate efficient in utero electroporation, which might also lead to a reduced number of animals.