Wang Hui, Matise Michael P
Department of Neuroscience and Cell Biology, UMDNJ-Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Methods Mol Biol. 2013;1018:133-40. doi: 10.1007/978-1-62703-444-9_13.
The developing spinal cord is a well-established model system widely used to study the signaling pathways and genetic programs that control neuronal/glial differentiation and neural circuit assembly. This is largely due to the relatively simple organization (compared to other CNS regions) and experimental accessibility of the neural tube, particularly in the chick embryo. In vivo transfection of cells within the developing chick neural tube using in ovo electroporation has emerged as a rapid and powerful experimental technique in that (1) transfected factors can be functionally tested in a spatially and temporally controlled manner and (2) the chick embryo provides a physiologically relevant in vivo environment to conduct biochemical studies such as dual-channel luciferase assay, co-immunoprecipitation (co-IP), and Chromatin Immunoprecipitation (ChIP). In this chapter, we will take an in-depth look at the in ovo electroporation system in embryonic chicken spinal cord. In the following chapter, we will continue by examining the use of in ovo electroporation in the dual-channel luciferase assay as an example of its biochemical application.
发育中的脊髓是一个成熟的模型系统,被广泛用于研究控制神经元/胶质细胞分化和神经回路组装的信号通路及遗传程序。这主要是由于神经管的组织结构相对简单(与其他中枢神经系统区域相比)且易于进行实验操作,尤其是在鸡胚中。利用胚内电穿孔对发育中的鸡神经管内的细胞进行体内转染,已成为一种快速且强大的实验技术,原因在于:(1)可以在空间和时间可控的方式下对转染因子进行功能测试;(2)鸡胚为进行诸如双通道荧光素酶测定、免疫共沉淀(co-IP)和染色质免疫沉淀(ChIP)等生化研究提供了生理相关的体内环境。在本章中,我们将深入探讨鸡胚脊髓中的胚内电穿孔系统。在下一章中,我们将继续以双通道荧光素酶测定中胚内电穿孔的应用为例,研究其生化应用。