使用地高辛标记RNA探针的小鼠胚胎全胚胎原位杂交

Whole-Mount In Situ Hybridization of Mouse Embryos Using DIG-Labeled RNA Probes.

作者信息

Wu Jingyi, Wang Xiaofang

机构信息

Department of Biomedical Sciences and Center for Craniofacial Research and Diagnosis, Texas A&M University College of Dentistry, Dallas, TX, USA.

Stomatological Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.

出版信息

Methods Mol Biol. 2019;1922:151-159. doi: 10.1007/978-1-4939-9012-2_15.

DOI:10.1007/978-1-4939-9012-2_15

PMID:30838573

Abstract

Whole-mount in situ hybridization (WMISH) is a commonly used technique for visualizing the expression profile of mRNAs in embryos. Unlike traditional in situ hybridization techniques, which require thin tissue sections, the WMISH technique allows gene expression patterns to be assessed over the entire embryo and structure. Here, we describe the detailed procedural steps of WMISH, including probe production, embryo fixation and staining, and post-hybridization signal detection. Using this protocol, we visualized highly specific expression patterns of Sonic hedgehog and Bmp4 mRNAs in E12.5 mouse embryos.

摘要

全胚胎原位杂交（WMISH）是一种用于观察胚胎中mRNA表达谱的常用技术。与需要薄组织切片的传统原位杂交技术不同，WMISH技术可在整个胚胎和结构上评估基因表达模式。在此，我们描述了WMISH的详细操作步骤，包括探针制备、胚胎固定和染色以及杂交后信号检测。使用该方案，我们在E12.5小鼠胚胎中观察到了音猬因子（Sonic hedgehog）和骨形态发生蛋白4（Bmp4）mRNA的高度特异性表达模式。