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Journal of Cell Science 114, 2855-2862 (2001)
© 2001 The Company of Biologists Limited

RESEARCH ARTICLE

Temporal relationship between cytochrome c release and mitochondrial swelling during UV-induced apoptosis in living HeLa cells

Wenhua Gao, Yongmei Pu, Kathy Q. Luo and Donald C. Chang*

Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, China
* Author for correspondence (e-mail: bochang{at}ust.hk )

Accepted May 2, 2001

During apoptosis, cytochrome c is released from mitochondria to the cytosol to activate a caspase cascade, which commits the cell to the death process. It has been proposed that the release of cytochrome c is caused by a swelling of the mitochondrial matrix triggered by the apoptotic stimuli. To test this theory, we measured directly the dynamic re-distribution of green fluorescence protein (GFP)-tagged cytochrome c and morphological change of mitochondria within living HeLa cells during u.v.-induced apoptosis. We observed that mitochondria did not swell when cytochrome c was released from mitochondria to cytosol during apoptosis. Instead, mitochondria swelled to spherical shapes within 10 minutes of cytochrome c release. This finding strongly suggests that cytochrome c release in apoptosis was not caused by mitochondrial swelling. This conclusion was further supported in two separated experiments using an immunostaining method and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) treatment. In addition, we found evidence that cytochrome c was also released before mitochondrial swelling in apoptosis induced by other cell death-inducing treatments, including tumor necrosis factor (TNF) and actinomycin D.

Key words: Apoptosis, Cytochrome c, Mitochondria, GFP, Programmed cell death


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