Measuring Apoptosis by Microscopy and Flow Cytometry.
Measuring Apoptosis by Microscopy and Flow Cytometry.
复制标题
DOI：
10.1002/0471142735.im1438s112
复制
发表时间：
2016-02-02
影响因子：
--
通讯作者：
Martin, Seamus J
中科院分区：
文献类型：
--
作者：
Hollville, Emilie;Martin, Seamus J
细胞凋亡是细胞程序性死亡的一种模式，在发育和维持组织稳态中起着重要作用。许多生理和病理刺激触发细胞凋亡，例如Fas、TRAIL或TNF受体的参与、生长因子剥夺、缺氧或暴露于细胞毒性药物。细胞凋亡由半胱氨酸蛋白酶的半胱天冬酶家族的成员从内部协调，半胱氨酸蛋白酶家族的成员在激活时触发一系列形态学变化，包括细胞收缩、广泛的质膜起泡、染色质凝聚、DNA水解和核碎裂。这些戏剧性的结构和生物化学的改变不仅导致细胞的受控解体，而且还导致吞噬细胞对凋亡细胞的有效识别和清除。坏死通常是非程序性的或通过压倒性的膜或细胞器损伤强加于细胞，其特征在于质膜快速破裂，随后是细胞器和细胞肿胀。坏死通常是由感染因子或严重偏离生理条件引起的。本单元描述了测量细胞凋亡和区分细胞凋亡与坏死的协议。
Apoptosis is a mode of programmed cell death that plays an important role during development and in the maintenance of tissue homeostasis. Numerous physiological as well as pathological stimuli trigger apoptosis such as engagement of Fas, TRAIL, or TNF receptors, growth factor deprivation, hypoxia, or exposure to cytotoxic drugs. Apoptosis is coordinated from within by members of the caspase family of cysteine proteases that, upon activation, trigger a series of morphological changes including cell shrinkage, extensive plasma membrane blebbing, chromatin condensation, DNA hydrolysis, and nuclear fragmentation. These dramatic structural and biochemical alterations result not only in the controlled dismantling of the cell, but also in the efficient recognition and removal of apoptotic cells by phagocytes. Necrosis, which is typically nonprogrammed or imposed upon the cell by overwhelming membrane or organelle damage, is characterized by rapid plasma membrane rupture followed by organelle and cell swelling. Necrosis is often provoked by infectious agents or a severe departure from physiological conditions. This unit describes protocols for the measurement of apoptosis and for distinguishing apoptosis from necrosis.