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Loss of cyclophilin D reveals a critical role for mitochondrial permeability transition in cell death

Abstract

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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Figure 1: Generation of Ppif-/- mice and assessment of mitochondrial pore function.
Figure 2: H2O2-induced mitochondrial dysfunction and death in Ppif-/- cells.
Figure 3: Cardiomyocyte death in Ppif-/- mice and CypD transgenic mice.
Figure 4: Cytochrome c release and cell death induced by Bax, tBid and Ca2+.

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References

  1. Halestrap, A. P., McStay, G. P. & Clarke, S. J. The permeability transition pore complex: another view. Biochimie 84, 153–166 (2002)

    Article  CAS  Google Scholar 

  2. Crompton, M. On the involvement of mitochondrial intermembrane junctional complexes in apoptosis. Curr. Med. Chem. 10, 1473–1484 (2003)

    Article  CAS  Google Scholar 

  3. Halestrap, A. P. Mitochondrial permeability: dual role of the ADP/ATP translocator? Nature 430, doi:10.1038/nature02816 (2004)

  4. Kokoszka, J. E. et al. The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore. Nature 427, 461–465 (2004)

    Article  ADS  CAS  Google Scholar 

  5. He, L. & Lemasters, J. J. Regulated and unregulated mitochondrial permeability transition pores: a new paradigm of pore structure and function? FEBS Lett. 512, 1–7 (2002)

    Article  ADS  CAS  Google Scholar 

  6. Scorrano, L. et al. A distinct pathway remodels mitochondrial cristae and mobilizes cytochrome c during apoptosis. Dev. Cell 2, 55–67 (2002)

    Article  CAS  Google Scholar 

  7. Zamzami, N. et al. Bid acts on the permeability transition pore complex to induce apoptosis. Oncogene 19, 6342–6350 (2000)

    Article  CAS  Google Scholar 

  8. Marzo, I. et al. Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. Science 281, 2027–2031 (1998)

    Article  ADS  CAS  Google Scholar 

  9. Pastorino, J. G., Chen, S. T., Tafani, M., Snyder, J. W. & Farber, J. L. The overexpression of Bax produces cell death upon induction of the mitochondrial permeability transition. J. Biol. Chem. 273, 7770–7775 (1998)

    Article  CAS  Google Scholar 

  10. Narita, M. et al. Bax interacts with the permeability transition pore to induce permeability transition and cytochrome c release in isolated mitochondria. Proc. Natl Acad. Sci. USA 95, 14681–14686 (1998)

    Article  ADS  CAS  Google Scholar 

  11. Shimizu, S., Narita, M. & Tsujimoto, Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399, 483–487 (1999)

    Article  ADS  CAS  Google Scholar 

  12. Eskes, R. et al. Bax-induced cytochrome C release from mitochondria is independent of the permeability transition pore but highly dependent on Mg2+ ions. J. Cell Biol. 143, 217–224 (1998)

    Article  CAS  Google Scholar 

  13. von Ahsen, O. et al. Preservation of mitochondrial structure and function after Bid- or Bax-mediated cytochrome c release. J. Cell Biol. 150, 1027–1036 (2000)

    Article  CAS  Google Scholar 

  14. Bossy-Wetzel, E., Newmeyer, D. D. & Green, D. R. Mitochondrial cytochrome c release in apoptosis occurs upstream of DEVD-specific caspase activation and independently of mitochondrial transmembrane depolarization. EMBO J. 17, 37–49 (1998)

    Article  CAS  Google Scholar 

  15. Griffiths, E. J. & Halestrap, A. P. Protection by Cyclosporin A of ischemia/reperfusion-induced damage in isolated rat hearts. J. Mol. Cell. Cardiol. 25, 1461–1469 (1993)

    Article  CAS  Google Scholar 

  16. Clarke, S. J., McStay, G. P. & Halestrap, A. P. Sanglifehrin A acts as a potent inhibitor of the mitochondrial permeability transition and reperfusion injury of the heart by binding to cyclophilin-D at a different site from cyclosporin A. J. Biol. Chem. 277, 34793–34799 (2002)

    Article  CAS  Google Scholar 

  17. Adams, J. W. et al. Cardiomyocyte apoptosis induced by Gαq signaling is mediated by permeability transition pore formation and activation of the mitochondrial death pathway. Circ. Res. 87, 1180–1187 (2000)

    Article  CAS  Google Scholar 

  18. Imberti, R., Nieminen, A. L., Herman, B. & Lemasters, J. J. Mitochondrial and glycolytic dysfunction in lethal injury to hepatocytes by t-butylhydroperoxide: protection by fructose, cyclosporin A and trifluoperazine. J. Pharmacol. Exp. Ther. 265, 392–400 (1993)

    CAS  PubMed  Google Scholar 

  19. Griffiths, E. J. & Halestrap, A. P. Mitochondrial non-specific pores remain closed during cardiac ischaemia, but open upon reperfusion. Biochem. J. 307, 93–98 (1995)

    Article  CAS  Google Scholar 

  20. Nieminen, A. L., Saylor, A. K., Tesfai, S. A., Herman, B. & Lemasters, J. J. Contribution of the mitochondrial permeability transition to lethal injury after exposure of hepatocytes to t-butylhydroperoxide. Biochem. J. 307, 99–106 (1995)

    Article  CAS  Google Scholar 

  21. Qian, T., Nieminen, A. L., Herman, B. & Lemasters, J. J. Mitochondrial permeability transition in pH-dependent reperfusion injury to rat hepatocytes. Am. J. Physiol. 273, C1783–C1792 (1997)

    Article  CAS  Google Scholar 

  22. Li, Y., Johnson, N., Capano, M., Edwards, M. & Crompton, M. Cyclophilin-D promotes the mitochondrial permeability transition but has opposite effects on apoptosis and necrosis. Biochem. J. 383, 101–109 (2004)

    Article  CAS  Google Scholar 

  23. Lin, D. T. & Lechleiter, J. D. Mitochondrial targeted cyclophilin D protects cells from cell death by peptidyl prolyl isomerization. J. Biol. Chem. 277, 31134–31141 (2002)

    Article  CAS  Google Scholar 

  24. Kaiser, R. A. et al. Targeted inhibition of p38 mitogen-activated protein kinase antagonizes cardiac injury and cell death following ischemia-reperfusion in vivo. J. Biol. Chem. 279, 15524–15530 (2004)

    Article  CAS  Google Scholar 

  25. Pallotti, F. & Lenaz, G. Isolation and subfractionation of mitochondria from animal cells and tissue culture lines. Methods Cell Biol. 65, 1–35 (2001)

    Article  CAS  Google Scholar 

  26. Petronilli, V. et al. The mitochondrial permeability transition, release of cytochrome c and cell death. Correlation with the duration of pore openings in situ. J. Biol. Chem. 276, 12030–12034 (2001)

    Article  CAS  Google Scholar 

  27. Kagawa, S. et al. A binary adenoviral vector system for expressing high levels of the proapoptotic gene bax. Gene Ther. 7, 75–79 (2000)

    Article  CAS  Google Scholar 

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Acknowledgements

@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.).

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Correspondence to Jeffery D. Molkentin.

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Supplementary information

Supplementary Figure S1

Mitochondrial swelling assays stimulated with low and high calcium. (PDF 75 kb)

Supplementary Figure S2

Assessment of calcium uptake content in purified mitochondria derived from wild-type hearts or Ppif null hearts. (PDF 99 kb)

Supplementary Figure S3

Phenotypic assessment of hearts from wild-type controls and Ppif heterozygous and null mice. (PDF 465 kb)

Supplementary Figure S4

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)

Supplementary Figure S5

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)

Supplementary Legends

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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