Reed Bruce, Harden Nicholas
Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON, Canada, N2L 3G1.
Department of Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, Canada, V5A 1S6.
Methods Mol Biol. 2017;1652:229-256. doi: 10.1007/978-1-4939-7219-7_16.
For several decades, genetic analysis in Drosophila has made important contributions to the understanding of signaling by Egfr. Egfr has been well characterized with regard to its oncogenic potential but is also being studied for its roles in organismal development. We have recently developed dorsal closure of the Drosophila embryo as a system for characterizing Egfr regulation of events that do not involve proliferation, as no cell divisions occur during this process. Dorsal closure is essentially a developmental wound healing event with parallels to vertebrate developmental epithelial fusions such as neural tube closure and palate fusion. We describe here a set of materials and protocols for studying Egfr signaling during dorsal closure, including assessing effects of altering Egfr signaling on other pathways, gene expression and, using live imaging, morphogenesis and programmed cell death. Although this "tool kit" is designed for looking at Egfr, it can be readily adapted to look at the participation of any signaling molecule in dorsal closure.
几十年来，果蝇的遗传分析为理解表皮生长因子受体（Egfr）信号传导做出了重要贡献。就其致癌潜力而言，Egfr已得到充分表征，但人们也在研究它在生物体发育中的作用。我们最近开发了果蝇胚胎背侧闭合作为一种系统，用于表征Egfr对不涉及增殖的事件的调控，因为在此过程中不发生细胞分裂。背侧闭合本质上是一种发育性伤口愈合事件，类似于脊椎动物发育过程中的上皮融合，如神经管闭合和腭融合。我们在此描述了一套用于研究背侧闭合过程中Egfr信号传导的材料和方案，包括评估改变Egfr信号传导对其他途径、基因表达的影响，以及使用实时成像研究形态发生和程序性细胞死亡。虽然这个“工具包”是为研究Egfr而设计的，但它可以很容易地改编用于研究任何信号分子在背侧闭合中的参与情况。
