Josefsson Emma C, White Michael J, Dowling Mark R, Kile Benjamin T
Cancer and Haematology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Methods Mol Biol. 2012;788:59-71. doi: 10.1007/978-1-61779-307-3_5.
Like many nucleated mammalian cells, the life and death of the anucleate platelet is regulated by Bcl-2 family proteins. Platelets depend on Bcl-x(L) for survival. Bcl-x(L) maintains platelet viability by restraining the killer protein Bak. When Bak is unleashed, it triggers classical intrinsic apoptosis by causing mitochondrial damage. The latter leads to caspase activation and phosphatidylserine (PS) exposure. Platelet apoptosis can be blocked by caspase inhibitors, or by genetic deletion of Bak and its close relative Bax. Perturbations in the platelet apoptosis program lead to changes in platelet life span in vivo. Here, we describe methods to determine platelet life span, enumerate young platelets, and measure hallmarks of platelet apoptosis, such as PS exposure, caspase activation, and mitochondrial dysfunction.
与许多有核哺乳动物细胞一样，无核血小板的生死受Bcl-2家族蛋白调控。血小板的存活依赖于Bcl-x(L)。Bcl-x(L)通过抑制杀伤蛋白Bak来维持血小板的活力。当Bak被释放时，它会通过造成线粒体损伤引发经典的内源性凋亡。后者导致半胱天冬酶激活和磷脂酰丝氨酸（PS）暴露。半胱天冬酶抑制剂或通过基因敲除Bak及其近亲Bax可阻断血小板凋亡。血小板凋亡程序的紊乱会导致体内血小板寿命的改变。在此，我们描述了测定血小板寿命、计数年轻血小板以及测量血小板凋亡特征（如PS暴露、半胱天冬酶激活和线粒体功能障碍）的方法。
