Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies
Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies
复制标题
DOI：
10.3791/52740
复制
发表时间：
2015-04-01
影响因子：
1.2
通讯作者：
Rubenstein, John L. R.
中科院分区：
文献类型：
--
作者：
Vogt, Daniel;Wu, Pei-Rung;Rubenstein, John L. R.
GABA能皮质中间神经元来源于胚胎内侧和尾侧神经节隆起(MGE和CGE)，在功能和形态上具有多样性。在理解不同的皮质中间神经元亚群的作用方面已经取得了进展，但仍有许多机制有待研究，这些机制可能有助于不同类型的GABA能细胞的发育和成熟。此外，GABA能信号的改变可能与自闭症、精神分裂症和癫痫的表型有关。特定的CRE驱动器系已经开始分离出独特的中间神经元亚群的功能。尽管在小鼠模型方面取得了进展，但用分子方法在体内有效地研究GABA能皮质间神经元祖细胞往往是困难的。研究这些细胞的细胞自主编程的一个重要技术是将MGE细胞移植到宿主大脑皮层。这些移植的细胞广泛迁移、分化和功能整合。此外，MGE细胞可以在移植前立即用慢病毒有效地转导，从而允许多种分子途径。在这里，我们详细介绍了一种协议，利用可用的Cre驱动系和Cre依赖的表达载体，在移植前有效地转导MGE细胞用于体内分析。这种方法是有利的，因为它结合了精确的遗传操作和这些细胞在移植后分散的能力，允许在体内获得更大的细胞类型特异性分辨率。
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.