Liu Kai-Li, Wang Xiu-Mei, Li Zi-Long, Liu Ying, He Rong-Qiao
State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
Methods Mol Biol. 2020;2047:363-375. doi: 10.1007/978-1-4939-9732-9_20.
The dynamic expression pattern analysis provides the primary information of gene function. Differences of the RNA and/or protein location will provide valuable information for gene expression regulation. Generally, in situ hybridization (ISH) and immunohistochemistry (IHC) are two main techniques to visualize the locations of gene transcripts and protein products in situ, respectively. Here we describe the protocol for the whole brain dissection, the in situ hybridization, and the immunostaining of the developing Xenopus brain sections. Additionally, we point out the modification of in situ hybridization for microRNA expression detection.
动态表达模式分析提供了基因功能的基本信息。RNA和/或蛋白质定位的差异将为基因表达调控提供有价值的信息。一般来说，原位杂交（ISH）和免疫组织化学（IHC）是分别在原位可视化基因转录本和蛋白质产物定位的两种主要技术。在这里，我们描述了非洲爪蟾发育中脑切片的全脑解剖、原位杂交和免疫染色的实验方案。此外，我们指出了用于微小RNA表达检测的原位杂交的改进方法。
