Report for: Avian and Reptilian Developmental Biology

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Chapter title	A Sensitive and Versatile In Situ Hybridization Protocol for Gene Expression Analysis in Developing Amniote Brains
Chapter number	22
Book title	Avian and Reptilian Developmental Biology
Published in	Methods in molecular biology, January 2017
DOI	10.1007/978-1-4939-7216-6_22
Pubmed ID	28809032
Book ISBNs	978-1-4939-7215-9, 978-1-4939-7216-6
Authors	Pei-Shan Hou, Takuma Kumamoto, Carina Hanashima
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.

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Avian and Reptilian Developmental Biology

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