Vogt Daniel, Wu Pei-Rung, Sorrells Shawn F, Arnold Christine, Alvarez-Buylla Arturo, Rubenstein John L R
Department of Psychiatry, University of California San Francisco;
Department of Psychiatry, University of California San Francisco.
J Vis Exp. 2015 Apr 23(98):52740. doi: 10.3791/52740.
GABAergic cortical interneurons, derived from the embryonic medial and caudal ganglionic eminences (MGE and CGE), are functionally and morphologically diverse. Inroads have been made in understanding the roles of distinct cortical interneuron subgroups, however, there are still many mechanisms to be worked out that may contribute to the development and maturation of different types of GABAergic cells. Moreover, altered GABAergic signaling may contribute to phenotypes of autism, schizophrenia and epilepsy. Specific Cre-driver lines have begun to parcel out the functions of unique interneuron subgroups. Despite the advances in mouse models, it is often difficult to efficiently study GABAergic cortical interneuron progenitors with molecular approaches in vivo. One important technique used to study the cell autonomous programming of these cells is transplantation of MGE cells into host cortices. These transplanted cells migrate extensively, differentiate, and functionally integrate. In addition, MGE cells can be efficiently transduced with lentivirus immediately prior to transplantation, allowing for a multitude of molecular approaches. Here we detail a protocol to efficiently transduce MGE cells before transplantation for in vivo analysis, using available Cre-driver lines and Cre-dependent expression vectors. This approach is advantageous because it combines precise genetic manipulation with the ability of these cells to disperse after transplantation, permitting greater cell-type specific resolution in vivo.
源自胚胎内侧和尾侧神经节隆起（MGE和CGE）的GABA能皮质中间神经元在功能和形态上具有多样性。在理解不同皮质中间神经元亚群的作用方面已经取得了进展，然而，仍有许多机制有待阐明，这些机制可能有助于不同类型GABA能细胞的发育和成熟。此外，GABA能信号改变可能导致自闭症、精神分裂症和癫痫的表型。特定的Cre驱动系已开始区分独特中间神经元亚群的功能。尽管小鼠模型取得了进展，但在体内用分子方法有效研究GABA能皮质中间神经元祖细胞往往很困难。用于研究这些细胞自主编程的一项重要技术是将MGE细胞移植到宿主皮质中。这些移植的细胞广泛迁移、分化并在功能上整合。此外，MGE细胞在移植前可立即用慢病毒高效转导，从而允许采用多种分子方法。在这里，我们详细介绍一种方案，利用现有的Cre驱动系和Cre依赖性表达载体，在移植前有效转导MGE细胞以进行体内分析。这种方法具有优势，因为它将精确的基因操作与这些细胞移植后分散的能力相结合，在体内实现更高的细胞类型特异性分辨率。
