期刊
JOURNAL OF CELL SCIENCE
卷 116, 期 8, 页码 1437-1447出版社
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.00320
关键词
apoptosis; mitochondria; photosensitisation; confocal microscopy; chloromethyl-X-rosamine
类别
We investigated the mode of signalling between mitochondria during apoptosis by monitoring the behaviour of non-irradiated mitochondria following microscopic photosensitisation of half the mitochondria in single human osteosarcoma cells loaded with CMXRos. Following partial irradiation of cells, non-irradiated mitochondria underwent a rapid depolarisation (within 10 minutes). The depolarisation was not inhibited by the caspase inhibitor zVAD-fmk but was suppressed by the intracellular Ca2+ chelator BAPTA and overexpression of Bcl-2. Significantly, such depolarisation occurred even after prior conversion of extended filamentous mitochondria into individual punctate structures, indicating that lumenal continuity is not required for communication between the irradiated and non-irradiated mitochondria. Partial irradiation of cells expressing cytochrome c-GFP revealed cytochrome c-GFP release from non-irradiated mitochondria at a delayed but unpredictable time interval (between 30 minutes and more than 2.5 hours) following irradiation, which was unaffected by zVAD-fmk. Once activated, cytochrome c-GFP release occurred within a 10 minute period. Inummocytochemistry failed to reveal the recruitment of Bax to non-irradiated mitochondria, which suggests that Bax does not mediate the release of cytochrome c from mitochondria. We conclude that signals (mediated by Ca2+) emanating from irradiated mitochondria are processed by their nonirradiated counterparts and comprise two temporally distinct phases, both independent of caspase-mediated amplification, which generate an initial rapid depolarisation and subsequent delayed release of cytochrome c.
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