Methods for the assessment of mitochondrial membrane permeabilization in apoptosis
Methods for the assessment of mitochondrial membrane permeabilization in apoptosis
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DOI：
10.1007/s10495-007-0720-1
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发表时间：
2007-05-01
期刊：
影响因子：
7.2
通讯作者：
Kroemer, Guido
中科院分区：
文献类型：
--
作者：
Galluzzi, Lorenzo;Zamzami, Naoufal;Kroemer, Guido
线粒体膜透化（MMP）被认为是许多程序性细胞死亡模型中的“不归点”。事实上，线粒体决定了细胞凋亡的内在途径，并且在外在途径中也起主要作用。MMP在不同程度上影响线粒体内膜和外膜（分别为IM和OM）。OM透化最终导致通常限制在线粒体膜间隙（IMS）中的蛋白质释放，包括半胱天冬酶激活剂（例如细胞色素c）和半胱天冬酶非依赖性死亡效应物（例如凋亡诱导因子）。部分IM透化破坏线粒体离子和体积稳态并消散线粒体跨膜电位（Δ Psi（m））。对早期线粒体改变的评估允许鉴定那些致力于死亡但尚未显示凋亡表型的细胞。已经开发了几种通过细胞荧光测定法和荧光显微镜测量MMP的技术。在这里，我们总结了目前可用的检测MMP的方法，并提供了这些技术的比较分析。
Mitochondrial membrane permeabilization (MMP) is considered as the "point-of-no-return" in numerous models of programmed cell death. Indeed, mitochondria determine the intrinsic pathway of apoptosis, and play a major role in the extrinsic route as well. MMP affects the inner and outer mitochondrial membranes (IM and OM, respectively) to a variable degree. OM permeabilization culminates in the release of proteins that normally are confined in the mitochondrial intermembrane space (IMS), including caspase activators (e.g. cytochrome c) and caspase-independent death effectors (e.g. apoptosis-inducing factor). Partial IM permeabilization disrupts mitochondrial ion and volume homeostasis and dissipates the mitochondrial transmembrane potential (Delta Psi(m)). The assessment of early mitochondrial alterations allows for the identification of cells that are committed to die but have not displayed yet the apoptotic phenotype. Several techniques to measure MMP by cytofluorometry and fluorescence microscopy have been developed. Here, we summarize the currently available methods for the detection of MMP, and provide a comparative analysis of these techniques.