Oxidative insults disrupt OPA1-mediated mitochondrial dynamics in cultured mammalian cells.
Oxidative insults disrupt OPA1-mediated mitochondrial dynamics in cultured mammalian cells.
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DOI：
10.1080/13510002.2018.1492766
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发表时间：
2018-12
期刊：
影响因子：
--
通讯作者：
Gilkerson R
中科院分区：
文献类型：
--
作者：
Garcia I;Innis-Whitehouse W;Lopez A;Keniry M;Gilkerson R
目的：探讨氧化损伤对线粒体动力学的影响。在哺乳动物细胞中，氧化损伤激活应激反应通路，包括炎症、细胞因子分泌和细胞凋亡。有趣的是，线粒体正在作为一个敏感的网络出现，它可能会作为后续细胞应激反应的早期指标。线粒体形成一个动态网络，平衡视神经萎缩-1(OPA1)介导的融合和动力蛋白相关蛋白-1(Drp1)介导的分裂事件，以维持动态平衡。方法：在这里，我们通过共聚焦显微镜、流式细胞术和基于蛋白质的分析，检测氧化损伤对143B骨肉瘤和H9c2心肌成肌细胞系线粒体动力学的影响。结果：在ROS供体过氧化氢(H_2O_2)作用下，两株细胞均表现出线粒体网络断裂和融合活性OPA1亚型丢失，表明OPA1介导的线粒体融合被氧化损伤所破坏。与此一致的是，缺乏OMA1的细胞可以保护自己免受OPA1的切割和线粒体断裂的影响，以响应H_2O_2的攻击。讨论：综上所述，这些发现表明，氧化损伤通过激活OMA1来破坏哺乳动物细胞中OPA1介导的线粒体动力学，这与线粒体动力学作为细胞应激信号的早期指标的新出现的作用是一致的。缩写：Δψm：跨膜电位；ROS：活性氧物种；过氧化氢：过氧化氢；OPA1：视神经萎缩-1；Mfn1：有丝分裂蛋白1；DRP1：动力蛋白相关蛋白1；DMEM：杜尔贝科改良的鹰培养基；PBS：磷酸盐缓冲液；TOM20：线粒体膜外膜转位酶-20；DAPI：二氨基苯吲哚；Tmre：四甲基罗丹明乙酯；tbst：三缓冲盐水吐温-20；MEF：小鼠胚胎成纤维细胞。
Objective: To explore the impact of oxidative insults on mitochondrial dynamics. In mammalian cells, oxidative insults activate stress response pathways including inflammation, cytokine secretion, and apoptosis. Intriguingly, mitochondria are emerging as a sensitive network that may function as an early indicator of subsequent cellular stress responses. Mitochondria form a dynamic network, balancing fusion, mediated by optic atrophy-1 (OPA1), and fission events, mediated by dynamin-related protein-1 (DRP1), to maintain homeostasis. Methods: Here, we examine the impact of oxidative insults on mitochondrial dynamics in 143B osteosarcoma and H9c2 cardiomyoblast cell lines via confocal microscopy, flow cytometry, and protein-based analyses. Results: When challenged with hydrogen peroxide (H2O2), a ROS donor, both cell lines display fragmentation of the mitochondrial network and loss of fusion-active OPA1 isoforms, indicating that OPA1-mediated mitochondrial fusion is disrupted by oxidative damage in mammalian cells. Consistent with this, cells lacking OMA1, a key protease responsible for cleavage of OPA1, are protected against OPA1 cleavage and mitochondrial fragmentation in response to H2O2 challenge. Discussion: Taken together, these findings indicate that oxidative insults damage OPA1-mediated mitochondrial dynamics in mammalian cells via activation of OMA1, consistent with an emerging role for mitochondrial dynamics as an early indicator of cellular stress signaling. Abbreviations: Δψm: transmembrane potential; ROS: reactive oxygen species; H2O2: hydrogen peroxide; OPA1: optic atrophy-1; MFN1: mitofusin1; DRP1: dynamin-related protein 1; DMEM: Dulbecco’s Modified Eagle’s Medium; PBS: phosphate buffer saline; TOM20: translocase of the outer mitochondrial membrane-20; DAPI: diaminophenylindole; TMRE: tetramethylrhodamine ethyl ester; TBST: Tris-Buffered Saline Tween-20; MEF: mouse embryonic fibroblast.
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影响因子：
64.8
作者：
Lee JE;Westrate LM;Wu H;Page C;Voeltz GK
通讯作者：
Voeltz GK
影响因子：
3.7
作者：
Moepert, Kristin;Hajek, Petr;Santel, Ansgar
通讯作者：
Santel, Ansgar
DOI：
10.1083/jcb.200211046
发表时间：
2003-01-20
期刊：
The Journal of cell biology
影响因子：
--
作者：
Chen H;Detmer SA;Ewald AJ;Griffin EE;Fraser SE;Chan DC
通讯作者：
Chan DC
影响因子：
2.7
作者：
Guillery, Olwenn;Malka, Florence;Rojo, Manuel
通讯作者：
Rojo, Manuel
DOI：
10.1083/jcb.200906084
发表时间：
2009-12-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ehses S;Raschke I;Mancuso G;Bernacchia A;Geimer S;Tondera D;Martinou JC;Westermann B;Rugarli EI;Langer T
通讯作者：
Langer T