Guglielmi G, De Renzis S
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Methods Cell Biol. 2017;139:167-186. doi: 10.1016/bs.mcb.2016.10.007. Epub 2016 Nov 23.
Morphogenesis of multicellular organisms is driven by changes in cell behavior, which happen at precise locations and defined developmental stages. Therefore, the studying of morphogenetic events would greatly benefit from tools that allow the perturbation of cell activity with spatial and temporal precision. We recently developed an optogenetic approach to modulate cell contractility with cellular precision and on fast (seconds) timescales during Drosophila embryogenesis. We present here a protocol to handle genetically engineered photosensitive Drosophila embryos and achieve light-mediated inhibition of apical constriction during tissue invagination. The possibility to modulate the levels of optogenetic activation at different laser powers makes this method suited also for studying how mechanical stresses are sensed and interpreted in vivo. Given the conserved function of cell contractility during animal development, the application of this method to other morphogenetic processes will facilitate our understanding of tissue mechanics and cell-cell interaction during morphogenesis.
多细胞生物的形态发生是由细胞行为的变化驱动的,这些变化发生在精确的位置和特定的发育阶段。因此,研究形态发生事件将极大地受益于能够在空间和时间上精确扰动细胞活性的工具。我们最近开发了一种光遗传学方法,可在果蝇胚胎发生过程中以细胞精度和快速(秒)时间尺度调节细胞收缩性。我们在此展示一种处理基因工程改造的光敏果蝇胚胎的方案,并在组织内陷过程中实现光介导的顶端收缩抑制。能够在不同激光功率下调节光遗传学激活水平,使得该方法也适用于研究体内机械应力是如何被感知和解读的。鉴于细胞收缩性在动物发育过程中的保守功能,将该方法应用于其他形态发生过程将有助于我们理解形态发生过程中的组织力学和细胞间相互作用。