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The physiological role of mitochondrial calcium revealed by mice lacking the mitochondrial calcium uniporter

Nature Cell Biology volume 15, pages 14641472 (2013) | Download Citation

Abstract

Mitochondrial calcium has been postulated to regulate a wide range of processes from bioenergetics to cell death. Here, we characterize a mouse model that lacks expression of the recently discovered mitochondrial calcium uniporter (MCU). Mitochondria derived from MCU−/− mice have no apparent capacity to rapidly uptake calcium. Whereas basal metabolism seems unaffected, the skeletal muscle of MCU−/− mice exhibited alterations in the phosphorylation and activity of pyruvate dehydrogenase. In addition, MCU−/− mice exhibited marked impairment in their ability to perform strenuous work. We further show that mitochondria from MCU−/− mice lacked evidence for calcium-induced permeability transition pore (PTP) opening. The lack of PTP opening does not seem to protect MCU−/− cells and tissues from cell death, although MCU−/− hearts fail to respond to the PTP inhibitor cyclosporin A. Taken together, these results clarify how acute alterations in mitochondrial matrix calcium can regulate mammalian physiology.

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References

  1. 1.

    & Calcium uptake by rat kidney mitochondria. Proc. Natl Acad. Sci. USA 47, 1744–1750 (1961).

  2. 2.

    & Ca ion uptake by rat kidney mitochondria and its dependence on respiration and phosphorylation. J. Biol. Chem. 237, 2670–2677 (1962).

  3. 3.

    & Mechanisms by which mitochondria transport calcium. Am. J. Physiol. 258, C755–C786 (1990).

  4. 4.

    & Characteristics and possible functions of mitochondrial Ca(2+) transport mechanisms. Biochim. Biophys. Acta 1787, 1291–1308 (2009).

  5. 5.

    , & The mitochondrial Ca(2+) uniporter. Cell Calcium 52, 16–21 (2012).

  6. 6.

    , & Calcium transport in mitochondria. FEBS Lett. 10, 1–5 (1970).

  7. 7.

    , & The mitochondrial calcium uniporter is a highly selective ion channel. Nature 427, 360–364 (2004).

  8. 8.

    Specific inhibition of mitochondrial Ca++ transport by ruthenium red. Biochem. Biophys. Res. Commun. 42, 298–305 (1971).

  9. 9.

    et al. Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476, 341–345 (2011).

  10. 10.

    , , , & A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 476, 336–340 (2011).

  11. 11.

    Regulation of mitochondrial dehydrogenases by calcium ions. Biochim. Biophys. Acta 1787, 1309–1316 (2009).

  12. 12.

    , & Mitochondrial transporters as novel targets for intracellular calcium signaling. Physiol. Rev. 87, 29–67 (2007).

  13. 13.

    , , & Ca(2+) activation of heart mitochondrial oxidative phosphorylation: role of the F(0)/F(1)-ATPase. Am. J. Physiol. Cell Physiol. 278, C423–C435 (2000).

  14. 14.

    Cardiac energy metabolism homeostasis: role of cytosolic calcium. J. Mol. Cell. Cardiol. 34, 1259–1271 (2002).

  15. 15.

    & Cellular metabolic homeostasis during large-scale change in ATP turnover rates in muscles. J. Exp. Biol. 200, 381–386 (1997).

  16. 16.

    & Ca2+ as a second messenger within mitochondria of the heart and other tissues. Annu. Rev. Phys. 52, 451–466 (1990).

  17. 17.

    & Role of mitochondrial Ca2+ in the regulation of cellular energetics. Biochemistry 51, 2959–2973 (2012).

  18. 18.

    & Regulation of mitochondrial Ca2+ and its effects on energetics and redox balance in normal and failing heart. J. Bioenerg. Biomembr. 41, 127–132 (2009).

  19. 19.

    & Mitochondrial permeability transition in Ca(2+)-dependent apoptosis and necrosis. Cell Calcium 50, 222–233 (2011).

  20. 20.

    , , & Mitochondria as sensors and regulators of calcium signalling. Nat. Rev. Mol. Cell Biol. 13, 566–578 (2012).

  21. 21.

    , & Regulation of cell death: the calcium-apoptosis link. Nat. Rev. Mol. Cell Biol. 4, 552–565 (2003).

  22. 22.

    , , & Mitochondrial calcium and the permeability transition in cell death. Biochim. Biophys. Acta 1787, 1395–1401 (2009).

  23. 23.

    , , & High- and low-calcium-dependent mechanisms of mitochondrial calcium signalling. Cell Calcium 44, 51–63 (2008).

  24. 24.

    & Myocardial ischemia-reperfusion injury: a neglected therapeutic target. J. Clin. Invest. 123, 92–100 (2013).

  25. 25.

    Mitochondrial calcium transport in the heart: physiological and pathological roles. J. Mol. Cell. Cardiol. 46, 789–803 (2009).

  26. 26.

    et al. Mitochondrial free calcium transients during excitation-contraction coupling in rabbit cardiac myocytes. FEBS Lett. 382, 31–36 (1996).

  27. 27.

    et al. MCUR1 is an essential component of mitochondrial Ca2+ uptake that regulates cellular metabolism. Nat. Cell Biol. 14, 1336–1343 (2012).

  28. 28.

    The energy-linked reaction of calcium with mitochondria. J. Biol. Chem. 240, 2729–2748 (1965).

  29. 29.

    , , & Characterization of the isozymes of pyruvate dehydrogenase phosphatase: implications for the regulation of pyruvate dehydrogenase activity. Biochim. Biophys. Acta 1652, 126–135 (2003).

  30. 30.

    & Regulation of pyruvate dehydrogenase complex activity by reversible phosphorylation. Biochem. Soc. Trans. 31, 1143–1151 (2003).

  31. 31.

    , & Screening assays for the mitochondrial permeability transition using a fluorescence multiwell plate reader. Anal. Biochem. 295, 220–226 (2001).

  32. 32.

    , & Cyclosporin A and cardioprotection: from investigative tool to therapeutic agent. Br. J. Pharmacol. 165, 1235–1245 (2012).

  33. 33.

    et al. Mice without myoglobin. Nature 395, 905–908 (1998).

  34. 34.

    et al. Skeletal muscles of mice deficient in muscle creatine kinase lack burst activity. Cell 74, 621–631 (1993).

  35. 35.

    et al. Living without creatine: unchanged exercise capacity and response to chronic myocardial infarction in creatine-deficient mice. Circ. Res. 112, 945–955 (2013).

  36. 36.

    et al. Mitochondrial calcium uniporter Mcu controls excitotoxicity and is transcriptionally repressed by neuroprotective nuclear calcium signals. Nat. Commun. 4, 2034 (2013).

  37. 37.

    , , , & Mitochondrial calcium uniporter silencing potentiates caspase-independent cell death in MDA-MB-231 breast cancer cells. Biochem. Biophys. Res. Commun. 434, 695–700 (2013).

  38. 38.

    , & Properties of a cyclosporin-insensitive permeability transition pore in yeast mitochondria. J. Biol. Chem. 272, 21104–21112 (1997).

  39. 39.

    et al. Bmi1 regulates mitochondrial function and the DNA damage response pathway. Nature 459, 387–392 (2009).

  40. 40.

    et al. Cysteine 203 of cyclophilin D is critical for cyclophilin D activation of the mitochondrial permeability transition pore. J. Biol. Chem. 286, 40184–40192 (2011).

  41. 41.

    , , & Dynamics of matrix-free Ca2+ in cardiac mitochondria: two components of Ca2+ uptake and role of phosphate buffering. J. Gen. Physiol. 139, 465–478 (2012).

  42. 42.

    et al. Resistance to Ca2+-induced opening of the permeability transition pore differs in mitochondria from glycolytic and oxidative muscles. Am. J. Physiol. Regul. Integr. Comp. Physiol. 295, R659–R668 (2008).

  43. 43.

    et al. Hypercontractile female hearts exhibit increased S-nitrosylation of the L-type Ca2+ channel alpha1 subunit and reduced ischemia/reperfusion injury. Circ. Res. 98, 403–411 (2006).

  44. 44.

    , , , & Protective effects of low and high doses of cyclosporin A against reoxygenation injury in isolated rat cardiomyocytes are associated with differential effects on mitochondrial calcium levels. Cell Calcium 27, 87–95 (2000).

  45. 45.

    et al. Mitochondrial matrix calcium is an activating signal for hormone secretion. Cell Metab. 13, 601–611 (2011).

  46. 46.

    et al. Atg7 modulates p53 activity to regulate cell cycle and survival during metabolic stress. Science 336, 225–228 (2012).

  47. 47.

    et al. p53 improves aerobic exercise capacity and augments skeletal muscle mitochondrial DNA content. Circ Res. 105, 705–712 (2009).

  48. 48.

    , , , & Preconditioning results in S-nitrosylation of proteins involved in regulation of mitochondrial energetics and calcium transport. Circ Res. 101, 1155–1163 (2007).

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Acknowledgements

We are grateful to C. Brantner, P. S. Connelly and M. P. Daniels of the NHLBI Electron Microscopy Core Facility for assistance with electron microscopy, C. Petucci of the Metabolomics Core Facility Sanford-Burnham Medical Research Institute for aiding in the metabolomic profiling, C. Combs and the NHLBI Microscopy Core for help with the Rhod-2 fluorescent measurements and A. Wiederkehr for the original mito-aequorin adenovirus. This work was supported by NIH Intramural funds.

Author information

Author notes

    • Xin Pan
    • , Jie Liu
    •  & Tiffany Nguyen

    These authors contributed equally to this work

Affiliations

  1. Center for Molecular Medicine, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Xin Pan
    • , Jie Liu
    • , Yanjie Teng
    • , Maria M. Fergusson
    • , Ilsa I. Rovira
    •  & Toren Finkel
  2. National Center of Biomedical Analysis, Beijing 100850, China

    • Xin Pan
  3. Systems Biology Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Tiffany Nguyen
    • , Junhui Sun
    • , Robert S. Balaban
    •  & Elizabeth Murphy
  4. iPSC and Genome Engineering Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Chengyu Liu
  5. Murine Phenotyping Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Michele Allen
    •  & Danielle A. Springer
  6. Proteomic Core, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Angel M. Aponte
    •  & Marjan Gucek

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Contributions

X.P., J.L. and T.N. designed, performed and analysed the experiments and aided in writing the manuscript, C.L. helped construct the mouse model, J.S., Y.T., M.M.F., I.I.R., M.A., D.A.S., A.M.A. and M.G. contributed to the completion of various experiments, and R.S.B., E.M. and T.F. conceived the study, supervised the research and contributed to writing the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Elizabeth Murphy or Toren Finkel.

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DOI

https://doi.org/10.1038/ncb2868

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