Chai Yongping, Zhu Zhiwen, Ou Guangshuo
Tsinghua-Peking Center for Life Sciences, School of Life Sciences and MOE Key Laboratory for Protein Science, Tsinghua University, Beijing, China.
Methods Mol Biol. 2018;1749:239-255. doi: 10.1007/978-1-4939-7701-7_18.
During C. elegans larval development, the Q neuroblasts produce their lineage by three rounds of divisions along with continuous cell migrations. Their neuronal progeny is dispersed from the pharynx to the anus. This in vivo system to study cell migration is appealing for several reasons. The lineage development is stereotyped; functional analysis and genomic screens are rendered easy and powerful thanks to powerful tools; transgenic manipulations and genome engineering are efficient and can be conveniently combined with live-cell imaging. Here we describe a series of protocols in Q cell migration studies, including quantifications of progeny position, genetic screening strategies, preparation of migration mutants or transgenic worms expressing related fluorescent proteins, multipositional time-lapse tracking of Q cell migration using confocal microscopy and image analyses of single cell movements and dynamics.
在秀丽隐杆线虫幼虫发育过程中,Q神经母细胞通过三轮分裂以及持续的细胞迁移产生其谱系。它们的神经元后代从咽部分散到肛门。这个用于研究细胞迁移的体内系统因多种原因而颇具吸引力。谱系发育是定型的;由于有强大的工具,功能分析和基因组筛选变得简单且强大;转基因操作和基因组工程效率高,并且可以方便地与活细胞成像相结合。在这里,我们描述了一系列Q细胞迁移研究的方案,包括后代位置的量化、遗传筛选策略、迁移突变体或表达相关荧光蛋白的转基因蠕虫的制备、使用共聚焦显微镜对Q细胞迁移进行多位置延时追踪以及单细胞运动和动态的图像分析。