Hou Pei-Shan, Kumamoto Takuma, Hanashima Carina
Laboratory for Neocortical Development, RIKEN Center for Developmental Biology, Kobe, 650-0047, Japan.
Sorbonne Universités, UPMC Univ Paris 06, INSERM U968, CNRS UMR 7210, Institut de la Vision, 17 rue Moreau, 75012, Paris, France.
Methods Mol Biol. 2017;1650:319-334. doi: 10.1007/978-1-4939-7216-6_22.
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.
在胚胎组织中检测特定RNA分子具有广泛的研究应用，包括研究大脑发育和进化过程中的基因表达动态。测序技术的最新进展引入了新的动物模型，以探索不同羊膜动物物种之间脑回路组装和多样化背后的分子原理。在这里，我们提供了一种通用原位杂交方法的详细步骤方案，该方法可立即应用于一系列羊膜动物胚胎，包括斑马雀和马达加斯加地壁虎，这两种新的模式生物近年来迅速崛起，用于比较脑研究。使用同一平台对从低丰度到高丰度表达范围的转录本进行灵敏检测，能够直接比较不同羊膜动物中感兴趣的基因，提供基因表达的高分辨率时空信息，以剖析大脑进化背后的分子原理。
