Protocol for analysis of senescent neuronal stem cells in genetic-modified embryonic mice using in utero electroporation technique

Liping Chen; Ying Li; Yan Wu; Shen Li; Xin Chang; Haitao Wu

doi:10.1016/j.xpro.2022.101461

Protocol for analysis of senescent neuronal stem cells in genetic-modified embryonic mice using in utero electroporation technique

STAR Protoc. 2022 Jun 13;3(3):101461. doi: 10.1016/j.xpro.2022.101461. eCollection 2022 Sep 16.

Authors

Liping Chen¹, Ying Li², Yan Wu², Shen Li², Xin Chang², Haitao Wu³

Affiliations

¹ Department of Neurobiology, Beijing Institute of Basic Medical Sciences, Beijing 100850, China. Electronic address: 2006443033@163.com.
² Department of Neurobiology, Beijing Institute of Basic Medical Sciences, Beijing 100850, China.
³ Department of Neurobiology, Beijing Institute of Basic Medical Sciences, Beijing 100850, China; Key Laboratory of Neuroregeneration, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu Province 226019, China; Chinese Institute for Brain Research, Beijing 102206, China. Electronic address: wuht@bmi.ac.cn.

Abstract

Here, we present a detailed protocol for identification and analysis of senescent neuronal stem cells (NSCs) in the developing mouse embryonic neocortex using in utero electroporation. We describe the procedures of in utero electroporation and brain slide preparation. We then detail the procedures of senescence-associated β-galactosidase (SA-β-Gal) staining and immunofluorescence staining, followed by microscope imaging analysis. This combined approach can be used to study the in situ senescence of in utero electroporated embryonic brains. For complete details on the use and execution of this protocol, please refer to Zhu et al. (2021).

Keywords: Cell Biology; Developmental biology; Microscopy; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Electroporation* / methods
Mice
Neocortex*
Neurons
Stem Cells