Oxidant stress evoked by pacemaking in dopaminergic neurons is attenuated by DJ-1

Article metrics

  • A Corrigendum to this article was published on 20 May 2015

Abstract

Parkinson’s disease is a pervasive, ageing-related neurodegenerative disease the cardinal motor symptoms of which reflect the loss of a small group of neurons, the dopaminergic neurons in the substantia nigra pars compacta1 (SNc). Mitochondrial oxidant stress is widely viewed as being responsible for this loss2, but why these particular neurons should be stressed is a mystery. Here we show, using transgenic mice that expressed a redox-sensitive variant of green fluorescent protein targeted to the mitochondrial matrix, that the engagement of plasma membrane L-type calcium channels during normal autonomous pacemaking created an oxidant stress that was specific to vulnerable SNc dopaminergic neurons. The oxidant stress engaged defences that induced transient, mild mitochondrial depolarization or uncoupling. The mild uncoupling was not affected by deletion of cyclophilin D, which is a component of the permeability transition pore, but was attenuated by genipin and purine nucleotides, which are antagonists of cloned uncoupling proteins. Knocking out DJ-1 (also known as PARK7 in humans and Park7 in mice), which is a gene associated with an early-onset form of Parkinson’s disease, downregulated the expression of two uncoupling proteins (UCP4 (SLC25A27) and UCP5 (SLC25A14)), compromised calcium-induced uncoupling and increased oxidation of matrix proteins specifically in SNc dopaminergic neurons. Because drugs approved for human use can antagonize calcium entry through L-type channels, these results point to a novel neuroprotective strategy for both idiopathic and familial forms of Parkinson’s disease.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Calcium influx through L-type calcium channels during pacemaking increases mitochondrial oxidant stress in SNc dopaminergic neurons.
Figure 2: Oxidant stress is elevated in SNc dopaminergic neurons from DJ-1 knockout mice.
Figure 3: Mitochondrial flickering is dependent on superoxide production and recruitment of mitochondrial uncoupling proteins.
Figure 4: Loss of DJ-1 attenuated UCP-dependent flickering in mitochondrial membrane potential.

References

  1. 1

    Albin, R. L., Young, A. B. & Penney, J. B. The functional anatomy of disorders of the basal ganglia. Trends Neurosci. 18, 63–64 (1995)

  2. 2

    Schapira, A. H. Mitochondria in the aetiology and pathogenesis of Parkinson’s disease. Lancet Neurol. 7, 97–109 (2008)

  3. 3

    Puopolo, M., Raviola, E. & Bean, B. P. Roles of subthreshold calcium current and sodium current in spontaneous firing of mouse midbrain dopamine neurons. J. Neurosci. 27, 645–656 (2007)

  4. 4

    Chan, C. S. et al. ‘Rejuvenation’ protects neurons in mouse models of Parkinson’s disease. Nature 447, 1081–1086 (2007)

  5. 5

    Khaliq, Z. M. & Bean, B. P. Pacemaking in dopaminergic ventral tegmental area neurons: depolarizing drive from background and voltage-dependent sodium conductances. J. Neurosci. 30, 7401–7413 (2010)

  6. 6

    Guzman, J. N., Sanchez-Padilla, J., Chan, C. S. & Surmeier, D. J. Robust pacemaking in substantia nigra dopaminergic neurons. J. Neurosci. 29, 11011–11019 (2009)

  7. 7

    Dooley, C. T. et al. Imaging dynamic redox changes in mammalian cells with green fluorescent protein indicators. J. Biol. Chem. 279, 22284–22293 (2004)

  8. 8

    Matlib, M. A. et al. Oxygen-bridged dinuclear ruthenium amine complex specifically inhibits Ca2+ uptake into mitochondria in vitro and in situ in single cardiac myocytes. J. Biol. Chem. 273, 10223–10231 (1998)

  9. 9

    Nicholls, D. G. & Ferguson, S. J. Bioenergetics 3 (Academic, 2002)

  10. 10

    Kahle, P. J., Waak, J. & Gasser, T. DJ-1 and prevention of oxidative stress in Parkinson’s disease and other age-related disorders. Free Radic. Biol. Med. 47, 1354–1361 (2009)

  11. 11

    Ehrenberg, B., Montana, V., Wei, M. D., Wuskell, J. P. & Loew, L. M. Membrane potential can be determined in individual cells from the Nernstian distribution of cationic dyes. Biophys. J. 53, 785–794 (1988)

  12. 12

    Rasola, A. & Bernardi, P. The mitochondrial permeability transition pore and its involvement in cell death and in disease pathogenesis. Apoptosis 12, 815–833 (2007)

  13. 13

    Krauss, S., Zhang, C. Y. & Lowell, B. B. The mitochondrial uncoupling-protein homologues. Nature Rev. Mol. Cell Biol. 6, 248–261 (2005)

  14. 14

    Brand, M. D. et al. Mitochondrial superoxide and aging: uncoupling-protein activity and superoxide production. Biochem. Soc. Symp. 71, 203–213 (2004)

  15. 15

    Andrews, Z. B. et al. Uncoupling protein-2 is critical for nigral dopamine cell survival in a mouse model of Parkinson’s disease. J. Neurosci. 25, 184–191 (2005)

  16. 16

    Zhang, C. Y. et al. Genipin inhibits UCP2-mediated proton leak and acutely reverses obesity- and high glucose-induced beta cell dysfunction in isolated pancreatic islets. Cell Metab. 3, 417–427 (2006)

  17. 17

    Echtay, K. S. et al. Superoxide activates mitochondrial uncoupling proteins. Nature 415, 96–99 (2002)

  18. 18

    Papa, S. & Skulachev, V. P. Reactive oxygen species, mitochondria, apoptosis and aging. Mol. Cell. Biochem. 174, 305–319 (1997)

  19. 19

    Canet-Aviles, R. M. et al. The Parkinson’s disease protein DJ-1 is neuroprotective due to cysteine-sulfinic acid-driven mitochondrial localization. Proc. Natl Acad. Sci. USA 101, 9103–9108 (2004)

  20. 20

    Bonifati, V. et al. Mutations in the DJ-1 gene associated with autosomal recessive early-onset Parkinsonism. Science 299, 256–259 (2003)

  21. 21

    Cookson, M. R. DJ-1, PINK1, and their effects on mitochondrial pathways. Mov. Disord. 25 (suppl. 1). S44–S48 (2010)

  22. 22

    Bender, A. et al. High levels of mitochondrial DNA deletions in substantia nigra neurons in aging and Parkinson disease. Nature Genet. 38, 515–517 (2006)

  23. 23

    Bender, A. et al. Dopaminergic midbrain neurons are the prime target for mitochondrial DNA deletions. J. Neurol. 255, 1231–1235 (2008)

  24. 24

    Krishnan, K. J., Greaves, L. C., Reeve, A. K. & Turnbull, D. M. Mitochondrial DNA mutations and aging. Ann. NY Acad. Sci. 1100, 227–240 (2007)

  25. 25

    Nicholls, D. G. Oxidative stress and energy crises in neuronal dysfunction. Ann. NY Acad. Sci. 1147, 53–60 (2008)

  26. 26

    Eisenberg, M. J., Brox, A. & Bestawros, A. N. Calcium channel blockers: an update. Am. J. Med. 116, 35–43 (2004)

  27. 27

    Becker, C., Jick, S. S. & Meier, C. R. Use of antihypertensives and the risk of Parkinson disease. Neurology 70, 1438–1444 (2008)

  28. 28

    Ritz, B. et al. L-type calcium channel blockers and Parkinson disease in Denmark. Ann. Neurol. 67, 600–606, 10.1002/ana.21937 (2010)

  29. 29

    Son, J. H. et al. Neuroprotection and neuronal differentiation studies using substantia nigra dopaminergic cells derived from transgenic mouse embryos. J. Neurosci. 19, 10–20 (1999)

  30. 30

    Bookout, A. L., Cummins, C. L., Mangelsdorf, D. J., Pesola, J. M. & Kramer, M. F. High-throughput real-time quantitative reverse transcription PCR. Curr. Protoc. Mol. Biol. Chapter 15, Unit 15 18. (2006)

  31. 31

    Schmittgen, T. D. & Livak, K. J. Analyzing real-time PCR data by the comparative C T method. Nature Protocols 3, 1101–1108 (2008)

  32. 32

    Tian, X., Kai, L., Hockberger, P. E., Wokosin, D. L. & Surmeier, D. J. MEF-2 regulates activity-dependent spine loss in striatopallidal medium spiny neurons. Mol. Cell. Neurosci. 44, 94–108 (2010)

  33. 33

    Fath, T., Ke, Y. D., Gunning, P., Gotz, J. & Ittner, L. M. Primary support cultures of hippocampal and substantia nigra neurons. Nature Protocols 4, 78–85 (2009)

Download references

Acknowledgements

We acknowledge the technical help of P. Hockberger, N. Schwarz, S. Ulrich, Y. Chen, C. S. Chan, D. Dryanovski and K. Saporito. We acknowledge S. Chan for supplying quantitative PCR primer sets. We acknowledge the gifts of DJ-1 knockout mice from T. and V. Dawson, Ucp2 knockout mice from D. Kong and B. Lowell, and cyclophilin D knockout mice from S. J. Korsmeyer. This work was supported by the Picower Foundation, the Hartman Foundation, the Falk Trust, the Parkinson’s Disease Foundation, NIH grants NS047085 (D.J.S.), NS 054850 (D.J.S.), K12GM088020 (J.S.-P.), HL35440 (P.T.S.) and RR025355 (P.T.S.), and DOD contract W81XWH-07-1-0170 (D.J.S.).

Author information

D.J.S. was responsible for the overall direction of the experiments, analysis of data, construction of figures and communication of the results. J.N.G. and J.S.-P. were responsible for the design and execution of experiments, as well as the analysis of results. D.W. provided expertise on optical approaches. E.I. conducted the immunocytochemical experiments. P.T.S. and J.K. were responsible for the generation of the TH-mito-roGFP mice; they also participated in the design, analysis and communication of the results.

Correspondence to D. James Surmeier.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

The file contains Supplementary Figures 1-7 with legends. (PDF 2278 kb)

Supplementary Movie 1

The movie shows TMRM fluorescence in an SNc dopaminergic neuron before and after bath application of isradipine (5 μM).Note the decreased flickering after application of isradipine. Similar results were seen in all of the neurons examined (n20). (MOV 12251 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Guzman, J., Sanchez-Padilla, J., Wokosin, D. et al. Oxidant stress evoked by pacemaking in dopaminergic neurons is attenuated by DJ-1. Nature 468, 696–700 (2010) doi:10.1038/nature09536

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.