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C57BL/6小鼠中皮质、海马、丘脑、下丘脑、外侧隔核和纹状体特异性的子宫内电穿孔

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse.

作者信息

Baumgart Jan, Baumgart Nadine

机构信息

TARC (Translational Animal Research Center), University Medical Center, JGU Mainz.

TARC (Translational Animal Research Center), University Medical Center, JGU Mainz;

出版信息

J Vis Exp. 2016 Jan 18(107):e53303. doi: 10.3791/53303.

Abstract

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.

摘要

子宫内电穿孔是一种广泛应用的技术,用于在啮齿动物中枢神经系统中对各种基因进行快速有效的时空操纵。与通过短发夹RNA(shRNA)介导的功能丧失研究一样,过表达所需基因也是可行的。因此,它具有广泛的应用范围。在特定区域靶向特定细胞的可行性进一步扩大了这种非常有用方法的潜在应用范围。为了有效地靶向特定区域,了解一些细微之处是必不可少的,例如胚胎阶段、施加的电压,最重要的是电极的位置。在这里,我们提供了一个详细的方案,可对C57BL/6小鼠中枢神经系统的几个区域进行特定且高效的子宫内电穿孔。特别展示了如何转染发育为压后皮质、运动皮质、体感皮质、梨状皮质、海马体1-3区、齿状回、纹状体、外侧隔核、丘脑和下丘脑的区域。为此,提供了有关合适胚胎阶段以及相应胚胎阶段合适电压的信息。最重要的是,描绘了一个角度图,该图指示了正极的合适位置。这个标准化方案有助于促进高效的子宫内电穿孔,这也可能导致动物数量的减少。

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本文引用的文献

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