Martin Emmanuel, Rocheleau-Leclair Olivier, Jenna Sarah
Department of Chemistry, Pharmaqam, Université du Québec à Montréal.
Department of Chemistry, Pharmaqam, Université du Québec à Montréal;
J Vis Exp. 2016 Mar 28(109):e53712. doi: 10.3791/53712.
Dissecting the signaling pathways that control the alteration of morphogenic processes during embryonic development requires robust and sensitive metrics. Embryonic elongation of the nematode Caenorhabditis elegans is a late developmental stage consisting of the elongation of the embryo along its longitudinal axis. This developmental stage is controlled by intercellular communication between hypodermal cells and underlying body-wall muscles. These signaling mechanisms control the morphology of hypodermal cells by remodeling the cytoskeleton and the cell-cell junctions. Measurement of embryonic lethality and developmental arrest at larval stages as well as alteration of cytoskeleton and cell-cell adhesion structures in hypodermal and muscle cells are classical phenotypes that have been used for more than 25 years to dissect these signaling pathways. Recent studies required the development of novel metrics specifically targeting either early or late elongation and characterizing morphogenic defects along the antero-posterior axis of the embryo. Here, we provide detailed protocols enabling the accurate measurement of the length and the width of the elongating embryos as well as the length of synchronized larvae. These methods constitute useful tools to identify genes controlling elongation, to assess whether these genes control both early and late phases of this stage and are required evenly along the antero-posterior axis of the embryo.
剖析在胚胎发育过程中控制形态发生过程改变的信号通路需要强大且灵敏的指标。线虫秀丽隐杆线虫的胚胎伸长是一个后期发育阶段,包括胚胎沿其纵轴伸长。这个发育阶段由皮下细胞与下方体壁肌肉之间的细胞间通讯控制。这些信号机制通过重塑细胞骨架和细胞间连接来控制皮下细胞的形态。测量胚胎致死率和幼虫阶段的发育停滞以及皮下和肌肉细胞中细胞骨架和细胞间粘附结构的改变是经典的表型,在超过25年的时间里一直被用于剖析这些信号通路。最近的研究需要开发新的指标,专门针对早期或晚期伸长,并表征沿胚胎前后轴的形态发生缺陷。在这里,我们提供了详细的方案,能够准确测量伸长胚胎的长度和宽度以及同步化幼虫的长度。这些方法是有用的工具,可用于鉴定控制伸长的基因,评估这些基因是否控制该阶段的早期和晚期,以及是否沿胚胎前后轴均匀发挥作用。