A Sensitive and Versatile In Situ Hybridization Protocol for Gene Expression Analysis in Developing Amniote Brains
A Sensitive and Versatile In Situ Hybridization Protocol for Gene Expression Analysis in Developing Amniote Brains
复制标题
一种灵敏且多功能的原位杂交方案，用于羊膜动物大脑发育中的基因表达分析
DOI：
10.1007/978-1-4939-7216-6_22
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Hanashima Carina
中科院分区：
文献类型：
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作者：
Hou Pei-Shan;Kumamoto Takuma;Hanashima Carina
胚胎组织中特异性RNA分子的检测具有广泛的研究应用，包括研究大脑发育和进化中的基因表达动态。测序技术的最新进展引入了新的动物模型，以探索不同物种之间脑回路组装和多样化的分子原理。在这里，我们提供了一个一步一步的协议，一个通用的原位杂交方法，可立即适用于一系列的胚胎，包括斑胸草雀和马达加斯加地壁虎，两个新的模式生物，已迅速出现的比较大脑研究在最近几年。使用相同的平台从低到高丰度表达范围的转录本的灵敏检测使得能够在不同的哺乳动物中直接比较感兴趣的基因，提供基因表达的高分辨率时空信息，以剖析大脑进化的分子原理。
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.