Mitochondria play a critical role in mediating both apoptotic and necrotic cell death. The mitochondrial permeability transition (mPT) leads to mitochondrial swelling, outer membrane rupture and the release of apoptotic mediators. The mPT pore is thought to consist of the adenine nucleotide translocator, a voltage-dependent anion channel, and cyclophilin D (the Ppif gene product), a prolyl isomerase located within the mitochondrial matrix1,2. Here we generated mice lacking Ppif and mice overexpressing cyclophilin D in the heart. Ppif null mice are protected from ischaemia/reperfusion-induced cell death in vivo, whereas cyclophilin D-overexpressing mice show mitochondrial swelling and spontaneous cell death. Mitochondria isolated from the livers, hearts and brains of Ppif null mice are resistant to mitochondrial swelling and permeability transition in vitro. Moreover, primary hepatocytes and fibroblasts isolated from Ppif null mice are largely protected from Ca2+-overload and oxidative stress-induced cell death. However, Bcl-2 family member-induced cell death does not depend on cyclophilin D, and Ppif null fibroblasts are not protected from staurosporine or tumour-necrosis factor-α-induced death. Thus, cyclophilin D and the mitochondrial permeability transition are required for mediating Ca2+- and oxidative damage-induced cell death, but not Bcl-2 family member-regulated death.
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@We would like to thank R. N. Kitsis for critical evaluation of this manuscript, M. E. Rothenberg, E. B. Brandt and S. P. Hogan for assistance with FACS analysis, M. B. Cohen and C. Liu for assistance in generating primary hepatocytes, and N. Zimmerman for help with mitochondrial Ca2+ uptake assays. This work was supported by Grants from the National Institutes of Health (J.D.M., J.R., G.W.D. and R.A.G.), by an American Heart Association Established Investigator Grant (J.D.M) and a National Institute of Health NRSA service award (R.A.K. and N.H.P.).
The authors declare that they have no competing financial interests.
Mitochondrial swelling assays stimulated with low and high calcium. (PDF 75 kb)
Assessment of calcium uptake content in purified mitochondria derived from wild-type hearts or Ppif null hearts. (PDF 99 kb)
Phenotypic assessment of hearts from wild-type controls and Ppif heterozygous and null mice. (PDF 465 kb)
Characterization of inner mitochondrial membrane potential with TMRE, in vivo mitochondrial pore opening with calcein-AM-CoCl2, and cell by PI nuclei staining, TUNEL, and annexin V labelling in wild-type and Ppif null embryonic fibroblasts. (PDF 1887 kb)
Phenotypic assessment of hearts from wild-type controls and cyclophilin D transgenic mice. Relative amount of caspase 9 cleavage and cytochrome c release from the heart is also shown. (PDF 552 kb)
Legends to accompany the above Supplementary Figures. (DOC 43 kb)
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Baines, C., Kaiser, R., Purcell, N. et al. Loss of cyclophilin D reveals a critical role for mitochondrial permeability transition in cell death. Nature 434, 658–662 (2005). https://doi.org/10.1038/nature03434
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