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{"title":"In Vivo Electroporation and Time-Lapse Imaging of the Rostral Migratory Stream in Developing Rodent Brain","authors":"Zhihui Huang, Ying Wang","doi":"10.1002/cpns.65","DOIUrl":null,"url":null,"abstract":"<p>Interneurons in the olfactory bulb are generated from neuronal precursor cells migrating from the anterior subventricular zone (SVZa) throughout the embryonic and postnatal life of mammals. This article describes basic methods for <i>in vivo</i> electroporation to label SVZa cells of both embryonic and postnatal rats. In addition, it describes three methods for tracing SVZa progenitors and following their migration pathway and differentiation, including immunohistochemistry, time-lapse live imaging in slice culture, and time-lapse imaging following transplantation in slice culture. These methods may be applied to all strains of rats and mice, including reporter mice. They may also be combined with methods such as BrdU labeling, tamoxifen injection, and electrophysiology, allowing one to observe proliferation or control gene expression at specific times and for specific neuronal functions. With time-lapse live imaging, details of labeled cells can be studied, including morphology, motility pattern, differentiation, and crosstalk between cells. © 2019 by John Wiley & Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.65","citationCount":"2","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Neuroscience","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpns.65","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Neuroscience","Score":null,"Total":0}
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Abstract
Interneurons in the olfactory bulb are generated from neuronal precursor cells migrating from the anterior subventricular zone (SVZa) throughout the embryonic and postnatal life of mammals. This article describes basic methods for in vivo electroporation to label SVZa cells of both embryonic and postnatal rats. In addition, it describes three methods for tracing SVZa progenitors and following their migration pathway and differentiation, including immunohistochemistry, time-lapse live imaging in slice culture, and time-lapse imaging following transplantation in slice culture. These methods may be applied to all strains of rats and mice, including reporter mice. They may also be combined with methods such as BrdU labeling, tamoxifen injection, and electrophysiology, allowing one to observe proliferation or control gene expression at specific times and for specific neuronal functions. With time-lapse live imaging, details of labeled cells can be studied, including morphology, motility pattern, differentiation, and crosstalk between cells. © 2019 by John Wiley & Sons, Inc.
发育中的啮齿动物大脑吻侧迁移流的体内电穿孔和延时成像
嗅球中的中间神经元是由哺乳动物在胚胎期和出生后从前脑室下区(SVZa)迁移过来的神经元前体细胞产生的。本文介绍了体外电穿孔标记胚胎和出生后大鼠SVZa细胞的基本方法。此外,还介绍了三种追踪SVZa祖细胞迁移途径和分化的方法,包括免疫组织化学、切片培养延时实时成像和切片培养移植后延时成像。这些方法适用于包括报告小鼠在内的所有大鼠和小鼠品系。它们也可以与BrdU标记、他莫昔芬注射和电生理学等方法相结合,允许在特定时间和特定神经元功能观察增殖或控制基因表达。通过延时实时成像,可以研究标记细胞的细节,包括形态、运动模式、分化和细胞间的串扰。©2019 by John Wiley &儿子,Inc。
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