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Characteristics of intracellular propagation of mitochondrial BAX recruitment during apoptosis
Apoptosis ( IF 6.1 ) Pub Date : 2021-01-11 , DOI: 10.1007/s10495-020-01654-w
Joshua A Grosser 1, 2 , Margaret E Maes 3 , Robert W Nickells 1, 4
Affiliation  

Recent advancements in live cell imaging technologies have identified the phenomenon of intracellular propagation of late apoptotic events, such as cytochrome c release and caspase activation. The mechanism, prevalence, and speed of apoptosis propagation remain unclear. Additionally, no studies have demonstrated propagation of the pro-apoptotic protein, BAX. To evaluate the role of BAX in intracellular apoptotic propagation, we used high speed live-cell imaging to visualize fluorescently tagged-BAX recruitment to mitochondria in four immortalized cell lines. We show that propagation of mitochondrial BAX recruitment occurs in parallel to cytochrome c and SMAC/Diablo release and is affected by cellular morphology, such that cells with processes are more likely to exhibit propagation. The initiation of propagation events is most prevalent in the distal tips of processes, while the rate of propagation is influenced by the 2-dimensional width of the process. Propagation was rarely observed in the cell soma, which exhibited near synchronous recruitment of BAX. Propagation velocity is not affected by mitochondrial volume in segments of processes, but is negatively affected by mitochondrial density. There was no evidence of a propagating wave of increased levels of intracellular calcium ions. Alternatively, we did observe a uniform increase in superoxide build-up in cellular mitochondria, which was released as a propagating wave simultaneously with the propagating recruitment of BAX to the mitochondrial outer membrane.



中文翻译:

细胞凋亡过程中线粒体 BAX 募集的细胞内传播特征

活细胞成像技术的最新进展已经确定了晚期凋亡事件的细胞内传播现象,例如细胞色素 c 释放和半胱天冬酶激活。细胞凋亡传播的机制、流行率和速度仍不清楚。此外,没有研究表明促凋亡蛋白 BAX 的增殖。为了评估 BAX 在细胞内凋亡增殖中的作用,我们使用高速活细胞成像来可视化四种永生化细胞系中荧光标记的 BAX 向线粒体的募集。我们表明,线粒体 BAX 募集的传播与细胞色素 c 和 SMAC/暗黑破坏神释放并行发生,并受细胞形态的影响,因此具有过程的细胞更有可能表现出传播。传播事件的启动在过程的远端最普遍,而传播速率受过程的二维宽度的影响。在细胞胞体中很少观察到繁殖,其表现出接近同步的 BAX 募集。传播速度不受过程片段中线粒体体积的影响,但受线粒体密度的负面影响。没有证据表明细胞内钙离子水平增加的传播波。或者,我们确实观察到细胞线粒体中超氧化物积聚的均匀增加,其作为传播波释放,同时 BAX 向线粒体外膜的传播募集。在细胞胞体中很少观察到繁殖,其表现出 BAX 的近同步募集。传播速度不受过程片段中线粒体体积的影响,但受线粒体密度的负面影响。没有证据表明细胞内钙离子水平增加的传播波。或者,我们确实观察到细胞线粒体中超氧化物积聚的均匀增加,其作为传播波释放,同时 BAX 向线粒体外膜的传播募集。在细胞胞体中很少观察到繁殖,其表现出接近同步的 BAX 募集。传播速度不受过程片段中线粒体体积的影响,但受线粒体密度的负面影响。没有证据表明细胞内钙离子水平增加的传播波。或者,我们确实观察到细胞线粒体中超氧化物积聚的均匀增加,其作为传播波释放,同时 BAX 向线粒体外膜的传播募集。没有证据表明细胞内钙离子水平增加的传播波。或者,我们确实观察到细胞线粒体中超氧化物积聚的均匀增加,其作为传播波释放,同时 BAX 向线粒体外膜的传播募集。没有证据表明细胞内钙离子水平增加的传播波。或者,我们确实观察到细胞线粒体中超氧化物积聚的均匀增加,其作为传播波释放,同时 BAX 向线粒体外膜的传播募集。

更新日期:2021-01-11
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