Hudson Clare
Laboratoire de Biologie du Développement de Villefranche-sur-mer (LBDV), Sorbonne Université, CNRS, Villefranche-sur-mer, France.
Methods Mol Biol. 2020;2047:325-345. doi: 10.1007/978-1-4939-9732-9_18.
The technique of in situ hybridization can be used to visualize the spatial and temporal pattern of gene expression during development. Ascidians are invertebrate chordates that develop with a fixed cell cleavage pattern into a tadpole larvae. The knowledge of the cell lineage allows the earliest steps of cell fate specification to be followed at a single cell resolution. This protocol describes preparation of Ciona intestinalis embryos, classical in situ hybridization protocol coupled with nuclear staining, and a guide to identify gene expression in specific precursors of the developing brain at neural plate stages of development.
原位杂交技术可用于观察发育过程中基因表达的时空模式。海鞘是无脊椎动物脊索动物，它们以固定的细胞分裂模式发育成蝌蚪幼虫。细胞谱系的知识使得能够以单细胞分辨率追踪细胞命运特化的最早步骤。本方案描述了玻璃海鞘胚胎的制备、结合核染色的经典原位杂交方案，以及在发育的神经板阶段识别发育中大脑特定前体细胞中基因表达的指南。
