In Situ Hybridization and Immunostaining of Xenopus Brain

Methods Mol Biol. 2020:2047:363-375. doi: 10.1007/978-1-4939-9732-9_20.

Abstract

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.

Keywords: Brain dissection; Expression pattern; Immunostaining; In situ hybridization; MicroRNA; Section; Xenopus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / embryology*
  • Brain / metabolism*
  • Immunohistochemistry
  • In Situ Hybridization / methods*
  • MicroRNAs / metabolism*
  • Xenopus laevis / embryology*
  • Xenopus laevis / metabolism*

Substances

  • MicroRNAs