Shklyar Boris, Shklover Jeny, Kurant Estee
Department of Anatomy and Cell Biology, Rappaport Institute for Research in the Medical Sciences, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology.
J Vis Exp. 2013 Aug 18(78):50151. doi: 10.3791/50151.
The proper elimination of unwanted or aberrant cells through apoptosis and subsequent phagocytosis (apoptotic cell clearance) is crucial for normal development in all metazoan organisms. Apoptotic cell clearance is a highly dynamic process intimately associated with cell death; unengulfed apoptotic cells are barely seen in vivo under normal conditions. In order to understand the different steps of apoptotic cell clearance and to compare 'professional' phagocytes--macrophages and dendritic cells to 'non-professional'--tissue-resident neighboring cells, in vivo live imaging of the process is extremely valuable. Here we describe a protocol for studying apoptotic cell clearance in live Drosophila embryos. To follow the dynamics of different steps in phagocytosis we use specific markers for apoptotic cells and phagocytes. In addition, we can monitor two phagocyte systems in parallel: 'professional' macrophages and 'semi-professional' glia in the developing central nervous system (CNS). The method described here employs the Drosophila embryo as an excellent model for real time studies of apoptotic cell clearance.
通过凋亡及随后的吞噬作用（凋亡细胞清除）适当清除不需要的或异常的细胞，对于所有后生动物的正常发育至关重要。凋亡细胞清除是一个与细胞死亡密切相关的高度动态过程；在正常条件下，体内几乎看不到未被吞噬的凋亡细胞。为了了解凋亡细胞清除的不同步骤，并将“专业”吞噬细胞——巨噬细胞和树突状细胞与“非专业”的组织驻留邻近细胞进行比较，对该过程进行体内实时成像极具价值。在这里，我们描述了一种在活的果蝇胚胎中研究凋亡细胞清除的方案。为了追踪吞噬作用不同步骤的动态变化，我们使用凋亡细胞和吞噬细胞的特异性标记物。此外，我们可以同时监测两个吞噬细胞系统：发育中的中枢神经系统（CNS）中的“专业”巨噬细胞和“半专业”神经胶质细胞。这里描述的方法将果蝇胚胎作为实时研究凋亡细胞清除的优秀模型。
