A sensitive and versatile in situ hybridization protocol for gene expression analysis in developing amniote brains

Pei Shan Hou, Takuma Kumamoto, Carina Hanashima

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

The detection of specific RNA molecules in embryonic tissues has wide research applications including studying gene expression dynamics in brain development and evolution. Recent advances in sequencing technologies have introduced new animal models to explore the molecular principles underlying the assembly and diversification of brain circuits between different amniote species. Here, we provide a step-by-step protocol for a versatile in situ hybridization method that is immediately applicable to a range of amniote embryos including zebra finch and Madagascar ground gecko, two new model organisms that have rapidly emerged for comparative brain studies over recent years. The sensitive detection of transcripts from low to high abundance expression range using the same platform enables direct comparison of gene of interest among different amniotes, providing high-resolution spatiotemporal information of gene expression to dissect the molecular principles underlying brain evolution.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages319-334
Number of pages16
Volume1650
DOIs
Publication statusPublished - 2017
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1650
ISSN (Print)1064-3745

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Keywords

  • Amniote embryos
  • Brain development
  • Dual color in situ hybridization
  • Evolution
  • Gecko
  • Pallium
  • Zebra finch

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Hou, P. S., Kumamoto, T., & Hanashima, C. (2017). A sensitive and versatile in situ hybridization protocol for gene expression analysis in developing amniote brains. In Methods in Molecular Biology (Vol. 1650, pp. 319-334). (Methods in Molecular Biology; Vol. 1650). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7216-6_22