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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Monitoring the mitochondrial transmembrane potential with the JC-1 fluorochrome in programmed cell death during mesophyll leaf senescence
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Monitoring the mitochondrial transmembrane potential with the JC-1 fluorochrome in programmed cell death during mesophyll leaf senescence

机译：在叶肉衰老过程中使用JC-1荧光染料监测程序性细胞死亡中的线粒体跨膜电位

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Analysis of the mitochondrial transmembrane potential (DeltaPsi(m)) with the help of the JC-1 fluorochrome (5,5',6,6'-tetrachloro-1.1',3,3'-tetraethylbenzimidazolcarbocyanine iodide) during mesophyll leaf senescence was performed in order to determine whether a reduction of DeltaPsi(m) takes place during mesophyll senescence and whether plant mitochondria. like mammalian ones, might be involved in the induction of programmed cell death. Fluorescence analysis of mesophyll protoplasts of Pisum sativum in a confocal microscope, fluorescent spectra analysis and time dependence of fluorescence intensity of monomers and of J-aggregates revealed that JC-1 is incorporated and accumulated specifically in plant mitochondria. Analysis of DeltaPsi(m) during mesophyll protoplast senescence revealed that two subpopulations of mitochondria which differ in DeltaPsi(m) exist in all analyzed stages of leaf senescence. The first subpopulation contains mitochondria with red fluorescence of J-aggregates due to an unperturbed high DeltaPsi(m). The second subpopulation comprises mitochondria with green fluorescence of monomers due to a low DeltaPsi(m). proving total depolarization of mitochondrial membranes. Fluorescence analysis demonstrated that even in the latest analyzed stages of leaf senescence. mitochondria with a high DeltaPsi(m) still exist. Fluorometric measurements revealed that the fluorescence intensity of J-aggregates decreases with the age of plants, which indicates that a reduction of DeltaPsi(m) during the mesophyll senescence process takes place; however. it does not take place within the whole population of mitochondria of the same protoplast. The reason of this can be due to a dramatic reorganization of mitochondria in mesophyll cells and the appearance of large mitochondria with local heterogeneity of DeltaPsi(m) in the oldest analyzed stages. All mitochondria in every stage of senescence maintained their membrane organization even when their size, distribution, and spatial organization in protoplasts changed dramatically. We stated that the reduction of DeltaPsi(m) does not directly induce programmed cell death in mesophyll cells, as opposed to animal apoptosis.
机译：在叶肉叶片衰老过程中，借助JC-1荧光染料（5,5'，6,6'-四氯-1.1'，3,3'-四乙基苯并咪唑碳菁碘）对线粒体跨膜电位（DeltaPsi（m））进行了分析。为了确定在叶肉衰老过程中是否发生DeltaPsi（m）的降低以及植物线粒体的表达，进行了检测。像哺乳动物一样，可能参与了程序性细胞死亡的诱导。在共聚焦显微镜下对豌豆叶肉原生质体的荧光分析，荧光光谱分析以及单体和J聚集体的荧光强度随时间的变化表明，JC-1专门掺入并积累在植物线粒体中。在叶肉原生质体衰老过程中对DeltaPsi（m）的分析表明，在所有叶片衰老的分析阶段，都有两个不同的线粒体亚群存在于DeltaPsi（m）中。第一个亚群包含线粒体，由于不受干扰的高DeltaPsi（m），J聚集体具有红色荧光。第二个亚群包括线粒体，由于低的DeltaPsi（m），具有绿色的单体荧光。证明线粒体膜完全去极化。荧光分析表明，即使在最近的叶片衰老分析阶段。高DeltaPsi（m）的线粒体仍然存在。荧光测定表明，J-聚集体的荧光强度随植物的年龄而降低，这表明在叶肉衰老过程中DeltaPsi（m）的降低。然而。它并不发生在同一原生质体的整个线粒体内。其原因可能是由于在最旧的分析阶段中叶肉细胞中线粒体的显着重组以及出现具有局部异质性的DeltaPsi（m）的大线粒体。即使原生质体的大小，分布和空间组织发生了巨大变化，衰老各个阶段中的所有线粒体仍保持其膜组织。我们说，与动物凋亡相反，减少DeltaPsi（m）不会直接诱导叶肉细胞中程序性细胞死亡。

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