Institute for Neurobiology, University of Münster, Germany.
Eur J Cell Biol. 2013 Oct-Nov;92(10-11):349-54. doi: 10.1016/j.ejcb.2013.09.003. Epub 2013 Oct 9.
Molecular understanding of actin dynamics requires a genetically traceable model system that allows live cell imaging together with high-resolution microscopy techniques. Here, we used Drosophila pupal macrophages that combine many advantages of cultured cells with a genetic in vivo model system. Using structured illumination microscopy together with advanced spinning disk confocal microscopy we show that these cells provide a powerful system for single gene analysis. It allows forward genetic screens to characterize the regulatory network controlling cell shape and directed cell migration in a physiological context. We knocked down components regulating lamellipodia formation, including WAVE, single subunits of Arp2/3 complex and CPA, one of the two capping protein subunits and demonstrate the advantages of this model system by imaging mutant macrophages ex vivo as well as in vivo upon laser-induced wounding.
肌动蛋白动力学的分子理解需要一个可遗传追踪的模型系统，该系统允许与高分辨率显微镜技术一起进行活细胞成像。在这里，我们使用果蝇蛹期巨噬细胞，将培养细胞的许多优势与体内遗传模型系统结合起来。我们使用结构照明显微镜和先进的旋转盘共聚焦显微镜，表明这些细胞为单基因分析提供了强大的系统。它允许正向遗传筛选来描述控制细胞形状和定向细胞迁移的调控网络，这是在生理环境下进行的。我们敲低了调节片状伪足形成的成分，包括 WAVE、Arp2/3 复合物的单个亚基和 CPA（两个盖帽蛋白亚基之一），通过对体外成像的突变巨噬细胞以及激光诱导损伤后的体内成像，展示了这个模型系统的优势。
