KINETICS OF PLASMA-MEMBRANE AND MITOCHONDRIAL ALTERATIONS IN CELLS UNDERGOING APOPTOSIS
KINETICS OF PLASMA-MEMBRANE AND MITOCHONDRIAL ALTERATIONS IN CELLS UNDERGOING APOPTOSIS
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DOI：
10.1002/cyto.990210308
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发表时间：
1995-11-01
期刊：
影响因子：
--
通讯作者：
REVILLARD, JP
中科院分区：
文献类型：
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作者：
LIZARD, G;FOURNEL, S;REVILLARD, JP
程序性细胞死亡或凋亡的特征在于典型的形态学改变。通过透射电子显微镜，凋亡细胞通过染色质与核膜紧密并置的浓缩、核膜的改变和核的碎裂来鉴定，而质膜和细胞器的完整性得以保留。相反，经历坏死的细胞显示出质膜的早期崩解和线粒体的肿胀，在这项研究中，我们通过流式细胞术评估了前向角光散射、90度光散射和与荧光素二乙酸酯、罗丹明123、和碘化丙啶在两个人B细胞系中进行拓扑异构酶II抑制剂VP-16诱导的细胞凋亡。将这些修饰的动力学与叠氮钠诱导的坏死细胞的动力学进行比较，在同一时间间隔，用Hoechst 33342染色后通过透射电子显微镜和紫外显微镜检查细胞，我们报告光散射和荧光素的顺序变化二醋酸酯在经历细胞凋亡或坏死的细胞中是相似的，而细胞凋亡的特征在于通过罗丹明123染色评估的线粒体活性的略微延迟的降低，令人惊讶的是，经历细胞凋亡的一部分细胞显示出碘化丙啶的早期摄取，随后是其核的浓缩和随后的碎裂，它的结论是碘化丙啶的摄取是一个非常早期的市场细胞死亡，不区分坏死和凋亡。沿着生物化学标准，Hoechst 33342染色显示的核形态学似乎是最简单和最具鉴别力的凋亡测定。(C)1995 Wiley-Liss，Inc.
Programmed cell death or apoptosis is characterized by typical morphological alterations. By transmission electron microscopy, apoptotic cells are identified by condensation of the chromatin in tight apposition to the nuclear envelope, alteration of the nuclear envelope and fragmentation of the nucleus, whereas integrity of the plasma membrane and organelles is preserved, Conversely cells undergoing necrosis display an early desintegration of cytoplasmic membrane and swelling of mitochondria, In this study we assessed by now cytometry the sequential alterations of forward angle Light scatter, 90 degrees light scatter, and fluorescence associated with fluorescein diacetate, rhodamine 123, and propidium iodide in two human B cell lines undergoing apoptosis induced by the topoisomerase II inhibitor VP-16. The kinetics of these modifications were compared to those of cells undergoing necrosis induced by sodium azide, At the same time intervals, cells were examined by transmission electron microscopy and by UV microscopy after staining with Hoechst 33342, We report that sequential changes in Light scatters and fluorescein diacetate are similar in cells undergoing apoptosis or necrosis, whereas apoptosis is characterized by a slightly delayed decrease of mitochondrial activity as assessed by rhodamine 123 staining, Surprisingly a part of cells undergoing apoptosis displayed an early uptake of propidium iodide followed by a condensation and then a fragmentation of their nuclei, It is concluded that uptake of propidium iodide is a very early market of cell death which does not discriminate between necrosis and apoptosis. Along with biochemical criteria, nuclear morphology revealed by staining with Hoechst 33342 would seem to be of the most simple and most discriminative assay of apoptosis. (C) 1995 Wiley-Liss, Inc.