Abstract

Synucleinopathies, such as Parkinson's disease and dementia with Lewy bodies, are neurodegenerative disorders that are characterized by the accumulation of α-synuclein (aSyn) in intracellular inclusions known as Lewy bodies. Prefibrillar soluble aSyn oligomers, rather than larger inclusions, are currently considered to be crucial species underlying synaptic dysfunction. We identified the cellular prion protein (PrPC) as a key mediator in aSyn-induced synaptic impairment. The aSyn-associated impairment of long-term potentiation was blocked in Prnp null mice and rescued following PrPC blockade. We found that extracellular aSyn oligomers formed a complex with PrPC that induced the phosphorylation of Fyn kinase via metabotropic glutamate receptors 5 (mGluR5). aSyn engagement of PrPC and Fyn activated NMDA receptor (NMDAR) and altered calcium homeostasis. Blockade of mGluR5-evoked phosphorylation of NMDAR in aSyn transgenic mice rescued synaptic and cognitive deficits, supporting the hypothesis that a receptor-mediated mechanism, independent of pore formation and membrane leakage, is sufficient to trigger early synaptic damage induced by extracellular aSyn.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & Synucleinopathies: common features and hippocampal manifestations. Cell. Mol. Life Sci. 74, 1485–1501 (2017).

  2. 2.

    , & Extracellular α-synuclein: a novel and crucial factor in Lewy body diseases. Nat. Rev. Neurol. 10, 92–98 (2014).

  3. 3.

    , , , & Cognitive status correlates with neuropathologic stage in Parkinson disease. Neurology 64, 1404–1410 (2005).

  4. 4.

    , , , & The spectrum of cognitive impairment in Lewy body diseases. Mov. Disord. 29, 608–621 (2014).

  5. 5.

    et al. Extracellular alpha-synuclein oligomers modulate synaptic transmission and impair LTP via NMDA-receptor activation. J. Neurosci. 32, 11750–11762 (2012).

  6. 6.

    et al. Adenosine A2A receptors modulate α-synuclein aggregation and toxicity. Cereb. Cortex 27, 718–730 (2017).

  7. 7.

    et al. The cellular prion protein mediates neurotoxic signalling of β-sheet-rich conformers independent of prion replication. EMBO J. 30, 2057–2070 (2011).

  8. 8.

    , , , & Cellular prion protein mediates impairment of synaptic plasticity by amyloid-β oligomers. Nature 457, 1128–1132 (2009).

  9. 9.

    , & The biological function of the cellular prion protein: an update. BMC Biol. 15, 34 (2017).

  10. 10.

    et al. Alzheimer amyloid-β oligomer bound to postsynaptic prion protein activates Fyn to impair neurons. Nat. Neurosci. 15, 1227–1235 (2012).

  11. 11.

    & Amyloid-β induced signaling by cellular prion protein and Fyn kinase in Alzheimer disease. Prion 7, 37–41 (2013).

  12. 12.

    , , & Membrane-bound beta-amyloid oligomers are recruited into lipid rafts by a fyn-dependent mechanism. FASEB J. 22, 1552–1559 (2008).

  13. 13.

    et al. Impaired long-term potentiation, spatial learning, and hippocampal development in fyn mutant mice. Science 258, 1903–1910 (1992).

  14. 14.

    & NMDA receptor subunits epsilon 1 (NR2A) and epsilon 2 (NR2B) are substrates for Fyn in the postsynaptic density fraction isolated from the rat brain. Biochem. Biophys. Res. Commun. 216, 582–588 (1995).

  15. 15.

    et al. Characterization of Fyn-mediated tyrosine phosphorylation sites on GluR epsilon 2 (NR2B) subunit of the N-methyl-D-aspartate receptor. J. Biol. Chem. 276, 693–699 (2001).

  16. 16.

    & Src kinases: a hub for NMDA receptor regulation. Nat. Rev. Neurosci. 5, 317–328 (2004).

  17. 17.

    et al. Molecular characterization and comparison of the components and multiprotein complexes in the postsynaptic proteome. J. Neurochem. 97 (Suppl. 1), 16–23 (2006).

  18. 18.

    et al. A progressive mouse model of Parkinson's disease: the Thy1-aSyn (“Line 61”) mice. Neurotherapeutics 9, 297–314 (2012).

  19. 19.

    et al. Cognitive deficits in a mouse model of pre-manifest Parkinson's disease. Eur. J. Neurosci. 35, 870–882 (2012).

  20. 20.

    et al. Amyloid-beta oligomers increase the localization of prion protein at the cell surface. J. Neurochem. 117, 538–553 (2011).

  21. 21.

    et al. Metabotropic glutamate receptor 5 is a coreceptor for Alzheimer aβ oligomer bound to cellular prion protein. Neuron 79, 887–902 (2013).

  22. 22.

    et al. Evolutionary expansion and anatomical specialization of synapse proteome complexity. Nat. Neurosci. 11, 799–806 (2008).

  23. 23.

    , , , & Adenosine A2A receptors permit mGluR5-evoked tyrosine phosphorylation of NR2B (Tyr1472) in rat hippocampus: a possible key mechanism in NMDA receptor modulation. J. Neurochem. 135, 714–726 (2015).

  24. 24.

    et al. Adenosine A(2A) receptor blockade reverts hippocampal stress-induced deficits and restores corticosterone circadian oscillation. Mol. Psychiatry 18, 320–331 (2013).

  25. 25.

    et al. Overexpression of adenosine A2A Receptors in rats: effects on depression, locomotion, and anxiety. Front. Psychiatry 5, 67 (2014).

  26. 26.

    et al. The caffeine-binding adenosine A2A receptor induces age-like HPA-axis dysfunction by targeting glucocorticoid receptor function. Sci. Rep. 6, 31493 (2016).

  27. 27.

    et al. Characterization of the potency, selectivity, and pharmacokinetic profile for six adenosine A2A receptor antagonists. Naunyn Schmiedebergs Arch. Pharmacol. 375, 133–144 (2007).

  28. 28.

    The synaptic pathology of α-synuclein aggregation in dementia with Lewy bodies, Parkinson's disease and Parkinson's disease dementia. Acta Neuropathol. 120, 131–143 (2010).

  29. 29.

    et al. Common structure of soluble amyloid oligomers implies common mechanism of pathogenesis. Science 300, 486–489 (2003).

  30. 30.

    et al. Formation of toxic oligomeric α-synuclein species in living cells. PLoS One 3, e1867 (2008).

  31. 31.

    & Alpha-synuclein: from secretion to dysfunction and death. Cell Death Dis. 3, e350 (2012).

  32. 32.

    et al. α-Synuclein oligomers oppose long-term potentiation and impair memory through a calcineurin-dependent mechanism: relevance to human synucleopathic diseases. J. Neurochem. 120, 440–452 (2012).

  33. 33.

    H. et al. Regulation of Amyloid β oligomer binding to neurons and neurotoxicity by the complex prion protein/mGluR5. J. Biol. Chem. 291, 21945–21955 (2016).

  34. 34.

    et al. Loss of prion protein leads to age-dependent behavioral abnormalities and changes in cytoskeletal protein expression. Mol. Neurobiol. 50, 923–936 (2014).

  35. 35.

    et al. Memory impairment in transgenic Alzheimer mice requires cellular prion protein. J. Neurosci. 30, 6367–6374 (2010).

  36. 36.

    et al. Prion protein attenuates excitotoxicity by inhibiting NMDA receptors. J. Cell Biol. 181, 551–565 (2008).

  37. 37.

    et al. Involvement of cellular prion protein in α-synuclein transport in neurons. Mol. Neurobiol. (2017).

  38. 38.

    et al. Physiology of the prion protein. Physiol. Rev. 88, 673–728 (2008).

  39. 39.

    & From cell protection to death: may Ca2+ signals explain the chameleonic attributes of the mammalian prion protein? Biochem. Biophys. Res. Commun. 379, 171–174 (2009).

  40. 40.

    et al. The complex PrP(c)-Fyn couples human oligomeric Aβ with pathological tau changes in Alzheimer's disease. J. Neurosci. 32, 16857–71a (2012).

  41. 41.

    et al. The prion protein constitutively controls neuronal store-operated Ca2+ entry through Fyn kinase. Front. Cell. Neurosci. 9, 416 (2015).

  42. 42.

    et al. Mice devoid of prion protein have cognitive deficits that are rescued by reconstitution of PrP in neurons. Neurobiol. Dis. 19, 255–265 (2005).

  43. 43.

    et al. Prion protein is necessary for normal synaptic function. Nature 370, 295–297 (1994).

  44. 44.

    , , , & Age-dependent loss of PTP and LTP in the hippocampus of PrP-null mice. Neurobiol. Dis. 13, 55–62 (2003).

  45. 45.

    et al. Impaired motor coordination in mice lacking prion protein. Cell. Mol. Neurobiol. 18, 731–742 (1998).

  46. 46.

    et al. Adenosine A2A receptors and metabotropic glutamate 5 receptors are co-localized and functionally interact in the hippocampus: a possible key mechanism in the modulation of N-methyl-D-aspartate effects. J. Neurochem. 95, 1188–1200 (2005).

  47. 47.

    & Istradefylline: first global approval. Drugs 73, 875–882 (2013).

  48. 48.

    & Adenosine A2A receptor gene disruption protects in an α-synuclein model of Parkinson's disease. Ann. Neurol. 71, 278–282 (2012).

  49. 49.

    et al. Increased α-synuclein levels in the cerebrospinal fluid of patients with Creutzfeldt-Jakob disease. J. Neurol. 261, 1203–1209 (2014).

  50. 50.

    et al. Common molecular mechanism of amyloid pore formation by Alzheimer's β-amyloid peptide and α-synuclein. Sci. Rep. 6, 28781 (2016).

  51. 51.

    et al. Differential neuropathological alterations in transgenic mice expressing alpha-synuclein from the platelet-derived growth factor and Thy-1 promoters. J. Neurosci. Res. 68, 568–578 (2002).

  52. 52.

    et al. Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein. Nature 356, 577–582 (1992).

  53. 53.

    , , & Place navigation impaired in rats with hippocampal lesions. Nature 297, 681–683 (1982).

  54. 54.

    et al. A2A adenosine receptor deletion is protective in a mouse model of tauopathy. Mol. Psychiatry 21, 97–107 (2016).

  55. 55.

    , , & Urocortin, but not urocortin II, protects cultured hippocampal neurons from oxidative and excitotoxic cell death via corticotropin-releasing hormone receptor type I. J. Neurosci. 22, 404–412 (2002).

  56. 56.

    et al. Neuroprotection afforded by adenosine A2A receptor blockade is modulated by corticotrophin-releasing factor (CRF) in glutamate injured cortical neurons. J. Neurochem. 123, 1030–1040 (2012).

  57. 57.

    et al. Twenty years of calcium imaging: cell physiology to dye for. Mol. Interv. 5, 112–127 (2005).

  58. 58.

    , , & Image analysis of Ca2+ signals as a basis for neurotoxicity assays: promises and challenges. Neurotoxicol. Teratol. 32, 16–24 (2010).

  59. 59.

    Histology of the central nervous system. Toxicol. Pathol. 39, 22–35 (2011).

  60. 60.

    et al. Heat-mediated enrichment of α-synuclein from cells and tissue for assessing post-translational modifications. J. Neurochem. 126, 673–684 (2013).

Download references

Acknowledgements

The authors thank L. Gros for figure layout design and C. Fahlbusch (University Medical Center Göttingen), A. Margarida Nascimento and J. Rino (Instituto de Medicina Molecular (iMM) Bioimaging facility), I. Moreira and J. Marques (iMM Rodent facility), and the iMM Histology and Comparative Pathology laboratory for technical assistance. M.T.F., H.V.M. and J.E.C. were supported by individual grants from Fundação para a Ciência e Tecnologia (FCT) (SFRH/BD/52228/2013; SFRH/BPD/109347/2015; SFRH/BPD/87647/2012); L.V.L. and T.F.O. were supported by a grant from the Fritz Thyssen Stiftung (Az. 10.12.2.165), Germany. L.V.L. received an iMM Lisboa internal fund (BIG – Breakthrough Idea Grant) for part of the project. L.V.L. is an Investigator FCT, Portugal. T.F.O. is supported by the DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Germany. LISBOA-01-0145-FEDER-007391, project co-financed by FEDER, POR Lisboa 2020 - Programa Operacional Regional de Lisboa, from PORTUGAL 2020 and by Fundação para a Ciência e a Tecnologia.

Author information

Author notes

    • Luísa V Lopes
    •  & Tiago F Outeiro

    These authors jointly directed this work.

Affiliations

  1. Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Universidade de Lisboa, Lisboa, Portugal.

    • Diana G Ferreira
    • , Mariana Temido-Ferreira
    • , Hugo Vicente Miranda
    • , Vânia L Batalha
    • , Joana E Coelho
    • , Inês Marques-Morgado
    • , Sandra H Vaz
    •  & Luísa V Lopes
  2. Department of Experimental Neurodegeneration, Center for Nanoscale Microscopy and Molecular Physiology of the Brain, Center for Biostructural Imaging of Neurodegeneration, University Medical Center Göttingen, Göttingen, Germany.

    • Diana G Ferreira
    • , Éva M Szegö
    •  & Tiago F Outeiro
  3. Department of Pharmacology and Therapeutics, Faculty of Medicine, University of Porto, Porto, Portugal; MedInUP - Center for Drug Discovery and Innovative Medicines, University of Porto, Porto, Portugal.

    • Diana G Ferreira
  4. CEDOC, Chronic Diseases Research Center, NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Lisboa, Portugal.

    • Hugo Vicente Miranda
    •  & Tiago F Outeiro
  5. Max Planck Institute for Experimental Medicine, Göttingen, Germany.

    • Jeong Seop Rhee
    •  & Tiago F Outeiro
  6. Department of Neurology, University Medical Center Göttingen, and German Center for Neurodegenerative Diseases (DZNE)-site Göttingen, Göttingen, Germany.

    • Matthias Schmitz
    •  & Inga Zerr

Authors

  1. Search for Diana G Ferreira in:

  2. Search for Mariana Temido-Ferreira in:

  3. Search for Hugo Vicente Miranda in:

  4. Search for Vânia L Batalha in:

  5. Search for Joana E Coelho in:

  6. Search for Éva M Szegö in:

  7. Search for Inês Marques-Morgado in:

  8. Search for Sandra H Vaz in:

  9. Search for Jeong Seop Rhee in:

  10. Search for Matthias Schmitz in:

  11. Search for Inga Zerr in:

  12. Search for Luísa V Lopes in:

  13. Search for Tiago F Outeiro in:

Contributions

D.G.F. performed most of the experimental work, analyzed data and wrote the manuscript. M.T.-F., J.E.C., V.L.B. and S.H.V. assisted with behavior and calcium experiments. E.M.S. assisted with animal experiments. I.M.-M. performed the immunohistochemistry experiments. M.S., J.S.R. and I.Z. provided the Prnp−/− mice and experimental support. H.V.M. produced and characterized aSyn species. L.V.L. and T.F.O. coordinated the study, designed the experiments and wrote the manuscript. All of the authors approved the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Luísa V Lopes or Tiago F Outeiro.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–7

  2. 2.

    Life Sciences Reporting Summary

Excel files

  1. 1.

    Supplementary Table 1

    Complete statistical details

Videos

  1. 1.

    aSyn oligomers increase intracellular Ca2+ in neurons

    Initial calcium response of FURA-2 AM loaded wildtype neurons to aSyn oligomers (aSyn mon, 500 nM)

  2. 2.

    aSyn oligomers do not increase intracellular Ca2+ in Prnp (-/-) neurons

    Initial calcium response of FURA-2 AM loaded Prnp (-/-) neurons to aSyn oligomers (aSyn mon, 500 nM)

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nn.4648

Further reading