School of Life Sciences, University of Nevada, Las Vegas, NV, USA.
Methods Mol Biol. 2021;2179:65-77. doi: 10.1007/978-1-0716-0779-4_8.
Epithelial-mesenchymal transitions (EMTs) drive the generation of cell diversity during both evolution and development. More and more evidence has pointed to a model where EMT is not a binary switch but a reversible process that can be stabilized at intermediate states. Despite our vast knowledge on the signaling pathways that trigger EMT, we know very little about how EMT happens in a step-wise manner. Live imaging of cells that are undergoing EMT in intact, living, animals will provide us valuable insights into how EMT is executed at both the cellular and molecular levels and help us identify and understand the intermediate states. Here, we describe how to image early stages of EMT in the mesoderm cells of live Drosophila melanogaster embryos and how to image contractile myosin that suspends EMT progression.
上皮-间充质转化(EMT)在进化和发育过程中驱动细胞多样性的产生。越来越多的证据表明,EMT 不是一个二元开关,而是一个可以在中间状态稳定的可逆过程。尽管我们对触发 EMT 的信号通路有了广泛的了解,但我们对 EMT 如何逐步发生知之甚少。对完整的、活体动物中正在经历 EMT 的细胞进行活细胞成像,将为我们提供 EMT 在细胞和分子水平上执行的宝贵见解,并帮助我们识别和理解中间状态。在这里,我们描述了如何在活体黑腹果蝇胚胎的中胚层细胞中对 EMT 的早期阶段进行成像,以及如何对暂停 EMT 进展的收缩性肌球蛋白进行成像。