Quinoprotein Adducts Accumulate in the Substantia Nigra of Aged Rats and Correlate with Dopamine-Induced Toxicity in SH-SY5Y Cells


Parkinson’s disease (PD) is an age-dependent neurodegenerative disorder characterized by dopaminergic neuron loss in substantia nigra. Previous studies have implicated a role of dopamine oxidation in PD. Dopamine oxidation leads to the formation of dopamine quinone, which generates reactive oxygen species and covalently modifies cysteinyl proteins to form quinoprotein adduct. We compared quinoprotein adduct formation and lipid peroxidation in different brain regions of young and old rats. We found a prominent age-dependent accumulation of quinoprotein adducts in the substantia nigra, while no significant change of lipid peroxidation was detected in any brain regions of 2- to 15-month old rats. To determine whether quinoprotein adduct formation correlates with dopamine-induced cytotoxicity, we analyzed dopamine treated SH-SY5Y cells and found a strong correlation between quinoprotein adduct formation and cytotoxicity. Together, our results indicate that quinoprotein adduct formation may play a role in the age-dependent selective vulnerability of dopaminergic neurons in PD.

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  1. 1.

    de Lau LM, Breteler MM (2006) Epidemiology of Parkinson’s disease. Lancet Neurol 5:525–535

    PubMed  Article  Google Scholar 

  2. 2.

    Nussbaum RL, Ellis CE (2003) Alzheimer’s disease and Parkinson’s disease. N Engl J Med 348:1356–1364

    PubMed  Article  CAS  Google Scholar 

  3. 3.

    Hastings TG, Lewis DA, Zigmond MJ (1996) Role of oxidation in the neurotoxic effects of intrastriatal dopamine injections. Proc Natl Acad Sci USA 93:1956–1961

    PubMed  Article  CAS  Google Scholar 

  4. 4.

    Colebrooke RE et al (2006) Age-related decline in striatal dopamine content and motor performance occurs in the absence of nigral cell loss in a genetic mouse model of Parkinson’s disease. Eur J Neurosci 24:2622–2630

    PubMed  Article  Google Scholar 

  5. 5.

    Caudle WM et al (2007) Reduced vesicular storage of dopamine causes progressive nigrostriatal neurodegeneration. J Neurosci 27:8138–8148

    PubMed  Article  CAS  Google Scholar 

  6. 6.

    Chen L et al (2008) Unregulated cytosolic dopamine causes neurodegeneration associated with oxidative stress in mice. J Neurosci 28:425–433

    PubMed  Article  Google Scholar 

  7. 7.

    Mosharov EV et al (2009) Interplay between cytosolic dopamine, calcium, and alpha-synuclein causes selective death of substantia nigra neurons. Neuron 62:218–229

    PubMed  Article  CAS  Google Scholar 

  8. 8.

    Eisenhofer G, Kopin IJ, Goldstein DS (2004) Catecholamine metabolism: a contemporary view with implications for physiology and medicine. Pharmacol Rev 56:331–349

    PubMed  Article  CAS  Google Scholar 

  9. 9.

    Caudle WM et al (2008) Altered vesicular dopamine storage in Parkinson’s disease: a premature demise. Trends Neurosci 31:303–308

    PubMed  Article  CAS  Google Scholar 

  10. 10.

    Graham DG (1978) Oxidative pathways for catecholamines in the genesis of neuromelanin and cytotoxic quinones. Mol Pharmacol 14:633–643

    PubMed  CAS  Google Scholar 

  11. 11.

    Sulzer D, Zecca L (2000) Intraneuronal dopamine-quinone synthesis: a review. Neurotox Res 1:181–195

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    Bolton JL et al (2000) Role of quinones in toxicology. Chem Res Toxicol 13:135–160

    PubMed  Article  CAS  Google Scholar 

  13. 13.

    Berman SB, Zigmond MJ, Hastings TG (1996) Modification of dopamine transporter function: effect of reactive oxygen species and dopamine. J Neurochem 67:593–600

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    Berman SB, Hastings TG (1999) Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria: implications for Parkinson’s disease. J Neurochem 73:1127–1137

    PubMed  Article  CAS  Google Scholar 

  15. 15.

    LaVoie MJ et al (2005) Dopamine covalently modifies and functionally inactivates parkin. Nat Med 11:1214–1221

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    Zigmond RE, Schwarzschild MA, Rittenhouse AR (1989) Acute regulation of tyrosine hydroxylase by nerve activity and by neurotransmitters via phosphorylation. Annu Rev Neurosci 12:415–461

    PubMed  Article  CAS  Google Scholar 

  17. 17.

    Haycock JW, Haycock DA (1991) Tyrosine hydroxylase in rat brain dopaminergic nerve terminals. Multiple-site phosphorylation in vivo and in synaptosomes. J Biol Chem 266:5650–5657

    PubMed  CAS  Google Scholar 

  18. 18.

    Zhang D et al (2007) Protein kinase C delta negatively regulates tyrosine hydroxylase activity and dopamine synthesis by enhancing protein phosphatase-2A activity in dopaminergic neurons. J Neurosci 27:5349–5362

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    Paz MA et al (1991) Specific detection of quinoproteins by redox-cycling staining. J Biol Chem 266:689–692

    PubMed  CAS  Google Scholar 

  20. 20.

    Tarozzi A et al (2009) Sulforaphane as an inducer of glutathione prevents oxidative stress-induced cell death in a dopaminergic-like neuroblastoma cell line. J Neurochem 111:1161–1171

    PubMed  Article  CAS  Google Scholar 

  21. 21.

    Snyder AM, Connor JR (2009) Iron, the substantia nigra and related neurological disorders. Biochim Biophys Acta 1790:606–614

    PubMed  CAS  Google Scholar 

  22. 22.

    Giovannelli L et al (2003) Vulnerability to DNA damage in the aging rat substantia nigra: a study with the comet assay. Brain Res 969:244–247

    PubMed  Article  CAS  Google Scholar 

  23. 23.

    Hattoria N et al (2009) Toxic effects of dopamine metabolism in Parkinson’s disease. Parkinsonism Relat Disord 15(Suppl 1):S35–S38

    PubMed  Article  Google Scholar 

  24. 24.

    Agil A et al (2006) Plasma lipid peroxidation in sporadic Parkinson’s disease. Role of the L-dopa. J Neurol Sci 240:31–36

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    Bernheimer H et al (1973) Brain dopamine and the syndromes of Parkinson and Huntington. Clinical, morphological and neurochemical correlations. J Neurol Sci 20:415–455

    PubMed  Article  CAS  Google Scholar 

  26. 26.

    Kanthasamy A et al (2010) Novel cell death signaling pathways in neurotoxicity models of dopaminergic degeneration: relevance to oxidative stress and neuroinflammation in Parkinson’s disease. Neurotoxicology 31:555–561

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    Hoyt KR, Reynolds IJ, Hastings TG (1997) Mechanisms of dopamine-induced cell death in cultured rat forebrain neurons: interactions with and differences from glutamate-induced cell death. Exp Neurol 143:269–281

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Berman SB, Hastings TG (1997) Inhibition of glutamate transport in synaptosomes by dopamine oxidation and reactive oxygen species. J Neurochem 69:1185–1195

    PubMed  Article  CAS  Google Scholar 

  29. 29.

    Kuhn DM, Arthur RE Jr (1999) L-DOPA-quinone inactivates tryptophan hydroxylase and converts the enzyme to a redox-cycling quinoprotein. Brain Res Mol Brain Res 73:78–84

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    Kuhn DM et al (1999) Tyrosine hydroxylase is inactivated by catechol-quinones and converted to a redox-cycling quinoprotein: possible relevance to Parkinson’s disease. J Neurochem 73:1309–1317

    PubMed  Article  CAS  Google Scholar 

  31. 31.

    Khan FH et al (2005) Inhibition of rat brain mitochondrial electron transport chain activity by dopamine oxidation products during extended in vitro incubation: implications for Parkinson’s disease. Biochim Biophys Acta 1741:65–74

    PubMed  Google Scholar 

  32. 32.

    Van Laar VS et al (2009) Proteomic identification of dopamine-conjugated proteins from isolated rat brain mitochondria and SH-SY5Y cells. Neurobiol Dis 34:487–500

    PubMed  Article  Google Scholar 

  33. 33.

    Khan FH, Saha M, Chakrabarti S (2001) Dopamine induced protein damage in mitochondrial-synaptosomal fraction of rat brain. Brain Res 895:245–249

    PubMed  Article  CAS  Google Scholar 

  34. 34.

    Jana S et al (2011) Mitochondrial dysfunction mediated by quinone oxidation products of dopamine: Implications in dopamine cytotoxicity and pathogenesis of Parkinson’s disease. Biochim Biophys Acta 1812:663–673

    PubMed  CAS  Google Scholar 

  35. 35.

    Conway KA et al (2001) Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct. Science 294:1346–1349

    PubMed  Article  CAS  Google Scholar 

  36. 36.

    Miyazaki I, Asanuma M (2009) Approaches to prevent dopamine quinone-induced neurotoxicity. Neurochem Res 34:698–706

    PubMed  Article  CAS  Google Scholar 

  37. 37.

    Miyazaki I et al (2007) Protective effects of metallothionein against dopamine quinone-induced dopaminergic neurotoxicity. FEBS Lett 581:5003–5008

    PubMed  Article  CAS  Google Scholar 

  38. 38.

    Asanuma M et al (2008) Preventing effects of a novel anti-parkinsonian agent zonisamide on dopamine quinone formation. Neurosci Res 60:106–113

    PubMed  Article  CAS  Google Scholar 

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This work was supported by the Ministry of Education of China project 985.

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Correspondence to Chonggang Yuan or Jiyan Ma.

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Wang, N., Wang, Y., Yu, G. et al. Quinoprotein Adducts Accumulate in the Substantia Nigra of Aged Rats and Correlate with Dopamine-Induced Toxicity in SH-SY5Y Cells. Neurochem Res 36, 2169 (2011). https://doi.org/10.1007/s11064-011-0541-z

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  • Parkinson disease
  • Dopamine
  • Dopamine quinone
  • Quinoprotein adduct
  • Lipid peroxidation
  • Aging