Alterations in mitochondrial structure and function are early events of dexamethasone-induced thymocyte apoptosis.
Alterations in mitochondrial structure and function are early events of dexamethasone-induced thymocyte apoptosis.
复制标题
线粒体结构和功能的改变是地塞米松诱导的胸腺细胞凋亡的早期事件。
DOI：
10.1083/jcb.130.1.157
复制
发表时间：
1995-07
影响因子：
7.8
通讯作者：
Gougeon, M L
中科院分区：
文献类型：
--
作者：
Petit, P X;Lecoeur, H;Zorn, E;Dauguet, C;Mignotte, B;Gougeon, M L
在本文中，我们使用了多参数的方法来分析广泛的胸腺细胞凋亡细胞死亡过程中发生的事件，此外，我们问是否在线粒体结构和功能的改变发生在细胞凋亡的早期阶段。对正常或凋亡胸腺细胞进行多参数定量分析，这些胸腺细胞是在培养基中或地塞米松存在下培养数小时后出现的。同时检测光散射特性，质膜的完整性（台盼蓝排斥），染色质凝聚（AO/EB染色的整个细胞或PI染色的细胞核），和DNA片段化（原位缺口翻译凋亡细胞）允许一个精确的分析的前凋亡和凋亡阶段。此外，对线粒体跨膜电位（Δ psi m）的全面研究在凋亡细胞摄取阳离子亲脂性染料DiOC 6（3）或J-聚集体形成阳离子JC-1的时间过程研究之后进行评估，表明Δ psi m的下降在胸腺细胞凋亡中非常早地发生，在DNA断裂之前。这与凋亡细胞中NAO摄取的细胞荧光研究评估的线粒体结构改变有关。最后，通过共聚焦和电子显微镜分析证实了在细胞凋亡的早期阶段发生的线粒体结构和功能的这些显著变化。
In this paper we used a multiparametric approach to analyze extensively the events occurring during apoptotic cell death of thymocytes, and furthermore, we asked whether alterations in mitochondrial structure and function are occurring in early stages of apoptosis. A multiparametric quantitative analysis was performed on normal or apoptotic thymocytes emerging from a few-hour culture performed in culture medium or in the presence of dexamethasone. Simultaneous detection of light scattering properties, integrity of plasma membrane (trypan blue exclusion), chromatin condensation (AO/EB staining of entire cells or PI staining of nuclei), and DNA fragmentation (in situ nick-translation in apoptotic cells) allowed a precise analysis of the preapoptotic and apoptotic stages. Moreover a thorough study of mitochondrial transmembrane potential (delta psi m) assessed following in a time course study the uptake by apoptotic cells of the cationic lipophilic dye DiOC6(3) or the J-aggregate-forming cation JC-1, indicates that a drop in delta psi m occurs very early in thymocyte apoptosis, before DNA fragmentation. This is associated with alteration in mitochondrial structure assessed by cytofluorimetric study of NAO uptake in apoptotic cells. Finally these dramatic alterations in mitochondrial structure and function occurring in early stages of apoptosis were confirmed by confocal and electron microscopy analysis.