Eimon Peter M
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.
Methods Enzymol. 2014;544:395-431. doi: 10.1016/B978-0-12-417158-9.00016-9.
Zebrafish (Danio rerio) have been extensively used to study apoptotic cell death during normal development and under a wide range of experimental manipulations. A number of features make zebrafish a particularly powerful model organism: (1) embryos are small in size, develop rapidly outside the mother, and are optically transparent; (2) tools are readily available for rapid knockdown and overexpression of genes; and (3) embryos can be arrayed into multiwell plates and are permeable to a wide range of drugs and small molecules. The molecular machinery underlying the intrinsic and extrinsic apoptosis pathways appears to be highly conserved between zebrafish and mammals. In this chapter, techniques are described for detecting apoptotic cells in situ in both fixed and live zebrafish embryos. Methods for inducing and inhibiting apoptosis and for functionally manipulating genes involved in apoptotic signaling are also discussed.
斑马鱼（Danio rerio）已被广泛用于研究正常发育过程中以及在各种实验操作下的凋亡细胞死亡。斑马鱼成为一种特别强大的模式生物有多个特点：（1）胚胎体积小，在母体外发育迅速，且具有光学透明性；（2）有现成的工具可用于快速敲低和过表达基因；（3）胚胎可排列在多孔板中，并且对多种药物和小分子具有通透性。斑马鱼与哺乳动物之间，内在和外在凋亡途径的分子机制似乎高度保守。在本章中，将描述在固定和活的斑马鱼胚胎中原位检测凋亡细胞的技术。还将讨论诱导和抑制凋亡以及在功能上操纵参与凋亡信号传导的基因的方法。
