Neurotoxicity Research

, Volume 19, Issue 4, pp 592–602

Neuroprotective Upregulation of Endogenous Alpha-Synuclein Precedes Ubiquitination in Cultured Dopaminergic Neurons



Alpha-synuclein is the major protein component of Lewy bodies—the pathological hallmark of Parkinson’s disease (PD) and Dementia with Lewy bodies (DLB). Its accumulation into intracellular aggregates is implicated in the process of Lewy body formation. However, its roles in both normal function, and disease, remain controversial. Using a novel model of chronic oxidative stress in cultured dopaminergic and cortical neurons, we report that endogenous alpha-synuclein is upregulated in response to low dose toxicity. This response is conserved between subpopulations of cortical and dopaminergic neurons, and confers relative resistance to apoptosis following secondary insult. Additional acute oxidative stress leads to intracellular accumulation of alpha-synuclein. These punctate deposits colocalize with ubiquitin, which is central to proteosome-mediated protein degeneration, and is the second major component of Lewy bodies. The current results imply that differential levels of alpha-synuclein expression may influence neuronal vulnerability in chronic neurodegenerative diseases. They further support a ‘two hit’ hypothesis for Lewy body formation, whereby mild stress causes a protective upregulation of alpha-synuclein. However, such increased levels of alpha-synuclein may drive its accumulation, following additional toxic insult. Finally, these results support a common mechanism for degeneration of dopaminergic and cortical neurons, affected in PD, and DLB, respectively.


Alpha-synuclein Oxidative stress Neuroprotection Dopaminergic Cortical Synucleinopathies 


  1. Baba M, Nakajo S, Tu PH, Tomita T, Nakaya K, Lee VM, Trojanowski JQ, Iwatsubo T (1998) Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies. Am J Pathol 152(4):879–884PubMedGoogle Scholar
  2. Betarbet R, Sherer TB, Greenamyre JT (2005) Ubiquitin-proteasome system and Parkinson’s diseases. Exp Neurol 191(Suppl 1):S17–S27PubMedCrossRefGoogle Scholar
  3. Bisaglia M, Mammi S, Bubacco L (2007) Kinetic and structural analysis of the early oxidation products of dopamine: analysis of the interactions with alpha-synuclein. J Biol Chem 282(21):15597–15600PubMedCrossRefGoogle Scholar
  4. Bisaglia M, Greggio E, Maric D, Miller DW, Cookson MR, Bubacco L (2010) Alpha-synuclein overexpression increases dopamine toxicity in BE2–M17 cells. BMC Neurosci 22:41CrossRefGoogle Scholar
  5. Cappai R, Leck SL, Tew DJ, Williamson NA, Smith DP, Galatis D, Sharples RA, Curtain CC, Ali FE, Cherny RA, Culvenor JG, Bottomley SP, Masters CL, Barnham KJ, Hill AF (2005) Dopamine promotes alpha-synuclein aggregation into SDS-resistant soluble oligomers via a distinct folding pathway. FASEB J 19(10):1377–1379PubMedGoogle Scholar
  6. Chartier-Harlin MC, Kachergus J, Roumier C, Mouroux V, Douay X, Lincoln S, Levecque C, Larvor L, Andrieux J, Hulihan M, Waucquier N, Defebvre L, Amouyel P, Farrer M, Destee A (2004) Alpha-synuclein locus duplication as a cause of familial Parkinson’s disease. Lancet 364(9440):1167–1169PubMedCrossRefGoogle Scholar
  7. Chesselet MF (2008) In vivo alpha-synuclein overexpression in rodents: a useful model of Parkinson’s disease? Exp Neurol 209(1):22–27PubMedCrossRefGoogle Scholar
  8. Cookson MR, van der Brug M (2008) Cell systems and the toxic mechanism(s) of alpha-synuclein. Exp Neurol 209(1):5–11PubMedCrossRefGoogle Scholar
  9. Cuervo AM, Stefanis L, Fredenburg R, Lansbury PT, Sulzer D (2004) Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy. Science 305(5688):1292–1295PubMedCrossRefGoogle Scholar
  10. Dickson TC, Adlard PA, Vickers JC (2000) Sequence of cellular changes following localized axotomy to cortical neurons in glia-free culture. J Neurotrauma 17(11):1095–1103PubMedCrossRefGoogle Scholar
  11. Engele J (1998) Spatial and temporal growth factor influences on developing midbrain dopaminergic neurons. J Neurosci Res 53(4):405–414PubMedCrossRefGoogle Scholar
  12. Feng L, Wang CY, Jiang H, Oho C, Mizuno K, Dugich-Djordjevic M, Lu B (1999) Differential effects of GDNF and BDNF on cultured ventral mesencephalic neurons. Brain Res Mol Brain Res 66(1–2):62–70PubMedCrossRefGoogle Scholar
  13. Gasser T (2007) Update on the genetics of Parkinson’s disease. Mov Disord 22(Suppl 17):S343–S350PubMedCrossRefGoogle Scholar
  14. Goldmann Gross R, Siderowf A, Hurtig HI (2008) Cognitive impairment in Parkinson’s disease and dementia with lewy bodies: a spectrum of disease. Neurosignals 16(1):24–34PubMedCrossRefGoogle Scholar
  15. Haas MA, Vickers JC, Dickson TC (2004) Binding partners L1 cell adhesion molecule and the ezrin–radixin–moesin (ERM) proteins are involved in development and the regenerative response to injury of hippocampal and cortical neurons. Eur J Neurosci 20(6):1436–1444PubMedCrossRefGoogle Scholar
  16. Hashimoto M, Hsu LJ, Xia Y, Takeda A, Sisk A, Sundsmo M, Masliah E (1999) Oxidative stress induces amyloid-like aggregate formation of NACP/alpha-synuclein in vitro. Neuroreport 10(4):717–721PubMedCrossRefGoogle Scholar
  17. Hashimoto M, Hsu LJ, Rockenstein E, Takenouchi T, Mallory M, Masliah E (2002) alpha-Synuclein protects against oxidative stress via inactivation of the c-Jun N-terminal kinase stress-signaling pathway in neuronal cells. J Biol Chem 277(13):11465–11472PubMedCrossRefGoogle Scholar
  18. Jellinger KA (2008) A critical reappraisal of current staging of Lewy-related pathology in human brain. Acta Neuropathol 116(1):1–16PubMedCrossRefGoogle Scholar
  19. King AE, Chung RS, Vickers JC, Dickson TC (2006) Localization of glutamate receptors in developing cortical neurons in culture and relationship to susceptibility to excitotoxicity. J Comp Neurol 498(2):277–294PubMedCrossRefGoogle Scholar
  20. Kowall NW, Hantraye P, Brouillet E, Beal MF, McKee AC, Ferrante RJ (2000) MPTP induces alpha-synuclein aggregation in the substantia nigra of baboons. Neuroreport 11(1):211–213PubMedCrossRefGoogle Scholar
  21. Leong SL, Pham CL, Galatis D, Fodero-Tavoletti MT, Perez K, Hill AF, Masters CL, Ali FE, Barnham KJ, Cappai R (2009) Formation of dopamine-mediated alpha-synuclein-soluble oligomers requires methionine oxidation. Free Radic Biol Med 46(10):1328–1337PubMedCrossRefGoogle Scholar
  22. Manning-Bog AB, McCormack AL, Purisai MG, Bolin LM, Di Monte DA (2003) Alpha-synuclein overexpression protects against paraquat-induced neurodegeneration. J Neurosci 23(8):3095–3099PubMedGoogle Scholar
  23. McGeer PL, McGeer EG (2008) The alpha-synuclein burden hypothesis of Parkinson disease and its relationship to Alzheimer disease. Exp Neurol 212(2):235–238PubMedCrossRefGoogle Scholar
  24. McKeith IG, Mosimann UP (2004) Dementia with Lewy bodies and Parkinson’s disease. Parkinsonism Relat Disord 10(Suppl 1):S15–S18PubMedCrossRefGoogle Scholar
  25. McNaught KS, Mytilineou C, Jnobaptiste R, Yabut J, Shashidharan P, Jennert P, Olanow CW (2002) Impairment of the ubiquitin-proteasome system causes dopaminergic cell death and inclusion body formation in ventral mesencephalic cultures. J Neurochem 81(2):301–306PubMedCrossRefGoogle Scholar
  26. Monti B, Polazzi E, Batti L, Crochemore C, Virgili M, Contestabile A (2007) Alpha-synuclein protects cerebellar granule neurons against 6-hydroxydopamine-induced death. J Neurochem 103(2):518–530PubMedCrossRefGoogle Scholar
  27. Morfis L, Cordato DJ (2006) Dementia with Lewy bodies in an elderly Greek male due to alpha-synuclein gene mutation. J Clin Neurosci 13(9):942–944PubMedCrossRefGoogle Scholar
  28. Outeiro TF, Lindquist S (2003) Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science 302(5651):1772–1775PubMedCrossRefGoogle Scholar
  29. Przedborski S (2005) Pathogenesis of nigral cell death in Parkinson’s disease. Parkinsonism Relat Disord 11(Suppl 1):S3–S7PubMedCrossRefGoogle Scholar
  30. Quilty MC, King AE, Gai WP, Pountney DL, West AK, Vickers JC, Dickson TC (2006) Alpha-synuclein is upregulated in neurones in response to chronic oxidative stress and is associated with neuroprotection. Exp Neurol 199(2):249–256PubMedCrossRefGoogle Scholar
  31. Richard IH, Papka M, Rubio A, Kurlan R (2002) Parkinson’s disease and dementia with Lewy bodies: one disease or two? Mov Disord 17(6):1161–1165PubMedCrossRefGoogle Scholar
  32. Rideout HJ, Dietrich P, Wang Q, Dauer WT, Stefanis L (2004) Alpha-synuclein is required for the fibrillar nature of ubiquitinated inclusions induced by proteasomal inhibition in primary neurons. J Biol Chem 279(45):46915–46920PubMedCrossRefGoogle Scholar
  33. Rideout HJ, Stefanis L (2002) Proteasomal inhibition-induced inclusion formation and death in cortical neurons require transcription and ubiquitination. Mol Cell Neurosci 21(2):223–238PubMedCrossRefGoogle Scholar
  34. Roy M, Sapolsky RM (2003) The exacerbation of hippocampal excitotoxicity by glucocorticoids is not mediated by apoptosis. Neuroendocrinology 77(1):24–31PubMedCrossRefGoogle Scholar
  35. Shults C (2006) Lewy bodies. Proc Natl Acad Sci USA 103:1661–1668PubMedCrossRefGoogle Scholar
  36. Singleton AB, Farrer M, Johnson J, Singleton A, Hague S, Kachergus J, Hulihan M, Peuralinna T, Dutra A, Nussbaum R, Lincoln S, Crawley A, Hanson M, Maraganore D, Adler C, Cookson MR, Muenter M, Baptista M, Miller D, Blancato J, Hardy J, Gwinn-Hardy K (2003) alpha-Synuclein locus triplication causes Parkinson’s disease. Science 302(5646):841PubMedCrossRefGoogle Scholar
  37. Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388(6645):839–840PubMedCrossRefGoogle Scholar
  38. Spillantini MG, Crowther RA, Jakes R, Hasegawa M, Goedert M (1998) alpha-Synuclein in filamentous inclusions of Lewy bodies from Parkinson’s disease and dementia with lewy bodies. Proc Natl Acad Sci USA 95(11):6469–6473PubMedCrossRefGoogle Scholar
  39. Wyllie AH, Kerr JF, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306PubMedCrossRefGoogle Scholar
  40. Xu J, Kao SY, Lee FJ, Song W, Jin LW, Yankner BA (2002) Dopamine-dependent neurotoxicity of alpha-synuclein: a mechanism for selective neurodegeneration in Parkinson disease. Nat Med 8(6):600–606PubMedCrossRefGoogle Scholar
  41. Zarranz JJ, Alegre J, Gomez-Esteban JC, Lezcano E, Ros R, Ampuero I, Vidal L, Hoenicka J, Rodriguez O, Atares B, Llorens V, Gomez Tortosa E, del Ser T, Munoz DG, de Yebenes JG (2004) The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 55(2):164–173PubMedCrossRefGoogle Scholar
  42. Zhou W, Schaack J, Zawada WM, Freed CR (2002) Overexpression of human alpha-synuclein causes dopamine neuron death in primary human mesencephalic culture. Brain Res 926(1–2):42–50PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • R. E. J. Musgrove
    • 1
  • A. E. King
    • 1
  • T. C. Dickson
    • 1
    • 2
  1. 1.Wicking Dementia Research and Education Centre, Menzies Research InstituteUniversity of TasmaniaHobartTasmaniaAustralia
  2. 2.Menzies Research InstituteUniversity of TasmaniaHobartAustralia

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