Kanaan Diana, Shklyar Boris, Porat-Kuperstein Lilach, Toledano Hila
Department of Human Biology, Faculty of Natural Sciences, University of Haifa, Haifa, Israel.
Bioimaging Unit, Faculty of Natural Sciences, University of Haifa, Haifa, Israel.
Bio Protoc. 2023 Mar 20;13(6):e4637. doi: 10.21769/BioProtoc.4637.
is a prevalent type of programmed cell death (PCD) in adult tissues in which phagocytes non-autonomously eliminate viable cells. Therefore, phagoptosis can only be studied in the context of the entire tissue that includes both the phagocyte executors and the targeted cells doomed to die. Here, we describe an ex vivo live imaging protocol of Drosophila testis to study the dynamics of phagoptosis of germ cell progenitors that are spontaneously removed by neighboring cyst cells. Using this approach, we followed the pattern of exogenous fluorophores with endogenously expressed fluorescent proteins and revealed the sequence of events in germ cell phagoptosis. Although optimized for Drosophila testis, this easy-to-use protocol can be adapted to a wide variety of organisms, tissues, and probes, thus providing a reliable and simple means to study phagoptosis.
是成年组织中一种普遍存在的程序性细胞死亡(PCD)类型,其中吞噬细胞非自主地清除存活细胞。因此,吞噬凋亡只能在包括吞噬细胞执行者和注定死亡的靶细胞的整个组织的背景下进行研究。在这里,我们描述了一种果蝇睾丸的体外实时成像方案,以研究生殖细胞祖细胞的吞噬凋亡动态,这些祖细胞被相邻的囊细胞自发清除。使用这种方法,我们追踪了外源性荧光团与内源性表达的荧光蛋白的模式,并揭示了生殖细胞吞噬凋亡中的事件顺序。尽管该方案是针对果蝇睾丸进行优化的,但这种易于使用的方案可以适用于多种生物体、组织和探针,从而为研究吞噬凋亡提供了一种可靠且简单的方法。