Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
International Research Center for Neurointelligence (WPI-IRCN), The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
STAR Protoc. 2021 May 17;2(2):100540. doi: 10.1016/j.xpro.2021.100540. eCollection 2021 Jun 18.
The embryonic mammalian neocortex includes neural progenitors and neurons at various stages of differentiation. The regulatory mechanisms underlying multiple aspects of neocortical development-including cell division, neuronal fate commitment, neuronal migration, and neuronal differentiation-have been explored using in utero electroporation and virus infection. Here, we describe a protocol for investigation of the effects of genetic manipulation on neural development through direct isolation of neural progenitors and neurons from the mouse embryonic neocortex by fluorescence-activated cell sorting. For complete details on the use and execution of this protocol, please refer to Tsuboi et al. (2018) and Sakai et al. (2019).
胚胎哺乳动物新皮质包含处于不同分化阶段的神经祖细胞和神经元。使用子宫内电穿孔和病毒感染，已经探索了新皮质发育的多个方面（包括细胞分裂、神经元命运决定、神经元迁移和神经元分化）的调控机制。在这里，我们描述了一种通过荧光激活细胞分选直接从小鼠胚胎新皮质中分离神经祖细胞和神经元来研究遗传操作对神经发育影响的方案。有关该方案使用和执行的完整详细信息，请参阅 Tsuboi 等人（2018 年）和 Sakai 等人（2019 年）。
