多细胞生物
生物
细胞生物学
细胞
吞噬作用
体内
表型
细胞凋亡
细胞命运测定
模式生物
体外
果蝇属(亚属)
施耐德2号电池
计算生物学
遗传学
基因
转录因子
RNA干扰
核糖核酸
作者
Elizabeth Silva,Jemima J. Burden,Nathalie C. Franc
出处
期刊:Methods in Enzymology
日期:2008-01-01
卷期号:: 39-59
被引量:5
标识
DOI:10.1016/s0076-6879(08)01603-0
摘要
Proper development of all multicellular organisms involves programmed apoptosis. Completion of this process requires removal of the resulting cell corpses through phagocytosis by their neighbors or by macrophages. Studies in C. elegans have been fruitful in the genetic dissection of key pathways, but they lack the professional immune system of higher organisms. Mammalian studies have identified a plethora of factors that are required for engulfment, but redundancy in the pathways has made it difficult to explain the genetic hierarchy of these factors. Thus, Drosophila has proven to be a useful evolutionary intermediate in which to examine this phenomenon. Here we describe methods used for dissecting the mechanisms and pathways involved in the engulfment of apoptotic cells by Drosophila phagocytes. Included are methods to be used for in vivo studies in the early embryo that can be used to examine engulfment of dying cells at various stages of embryogenesis. We also describe in vitro techniques for the use of Drosophila cell culture, including cell engulfment assays, that can be used for general phenotypic analysis, as well as live cell studies. We provide advice on imaging, including the preparation of samples for high-resolution microscopy and quantification of potential engulfment phenotypes for both in vivo and in vitro methods.
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