Advertisement

Prion Efficiently Replicates in α-Synuclein Knockout Mice

  • Edoardo Bistaffa
  • Martina Rossi
  • Chiara Maria Giulia De Luca
  • Federico Cazzaniga
  • Olga Carletta
  • Ilaria Campagnani
  • Fabrizio Tagliavini
  • Giuseppe Legname
  • Giorgio Giaccone
  • Fabio ModaEmail author
Article

Abstract

Prion diseases are a group of neurodegenerative disorders associated with the conformational conversion of the cellular prion protein (PrPC) into an abnormal misfolded form named PrPSc. Other than accumulating in the brain, PrPSc can bind PrPC and force it to change conformation to PrPSc. The exact mechanism which underlies the process of PrPC/PrPSc conversion still needs to be defined and many molecules or cofactors might be involved. Several studies have documented an important role of PrPC to act as receptor for abnormally folded forms of α-synuclein which are responsible of a group of diseases known as synucleinopathies. The presence of PrPC was required to promote efficient internalization and spreading of abnormal α-synuclein between cells. In this work, we have assessed whether α-synuclein exerts any role in PrPSc conversion and propagation either in vitro or in vivo. Indeed, understanding the mechanism of PrPC/PrPSc conversion and the identification of cofactors involved in this process is crucial for developing new therapeutic strategies. Our results showed that PrPSc was able to efficiently propagate in the brain of animals even in the absence of α-synuclein thus suggesting that this protein did not act as key modulator of prion propagation. Thus, α-synuclein might take part in this process but is not specifically required for sustaining prion conversion and propagation.

Keywords

Prions RML α-Synuclein PMCA 

Notes

Acknowledgments

The authors wish to thank Associazione Italiana Encefalopatie da Prioni (AIEnP).

Author Contributions

E.B. performed most of the experiments; M.R., C.D.L., performed part of the biochemical analysis; F.C., O.C. performed part of the histological analysis; I.C. was in charge of the animal care and sacrifice; F.T. and G.L. contributed in planning the experiments and in analyzing the data; and G.G. and F.M. supervised the work, analyzed the data and prepared the final version of the manuscript. All the authors read and approved the final manuscript.

Funding

This work was supported by the Italian Ministry of Health (RC) to F.M., Italian Ministry of Health to F.T., and Associazione Italiana Encefalopatie da Prioni (AIEnP).

Compliance with Ethical Standards

The study, including its Ethics aspects, was approved by the Italian Ministry of Health (Permit Number, NP-02-13).

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12035_2019_1602_MOESM1_ESM.jpg (632 kb)
ESM 1 (JPG 631 kb)

References

  1. 1.
    Chesebro B (1999) Prion protein and the transmissible spongiform encephalopathy diseases. Neuron 24(3):503–506CrossRefGoogle Scholar
  2. 2.
    Aguzzi A (2006) Prion diseases of humans and farm animals: epidemiology, genetics, and pathogenesis. J Neurochem 97(6):1726–1739CrossRefGoogle Scholar
  3. 3.
    Wulf MA, Senatore A, Aguzzi A (2017) The biological function of the cellular prion protein: an update. BMC Biol 15(1):34CrossRefGoogle Scholar
  4. 4.
    Prusiner SB (1998) Prions. Proc Natl Acad Sci U S A 95(23):13363–13383CrossRefGoogle Scholar
  5. 5.
    Bueler H, Aguzzi A, Sailer A, Greiner RA, Autenried P, Aguet M, Weissmann C (1993) Mice devoid of PrP are resistant to scrapie. Cell 73(7):1339–1347CrossRefGoogle Scholar
  6. 6.
    Prusiner SB, Scott M, Foster D, Pan KM, Groth D, Mirenda C, Torchia M, Yang SL et al (1990) Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63(4):673–686CrossRefGoogle Scholar
  7. 7.
    McKinley MP, Bolton DC, Prusiner SB (1983) A protease-resistant protein is a structural component of the scrapie prion. Cell 35(1):57–62CrossRefGoogle Scholar
  8. 8.
    Pan KM, Baldwin M, Nguyen J, Gasset M, Serban A, Groth D, Mehlhorn I, Huang Z et al (1993) Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci U S A 90(23):10962–10966CrossRefGoogle Scholar
  9. 9.
    Khalili-Shirazi A, Summers L, Linehan J, Mallinson G, Anstee D, Hawke S, Jackson GS, Collinge J (2005) PrP glycoforms are associated in a strain-specific ratio in native PrPSc. J Gen Virol 86(Pt 9):2635–2644CrossRefGoogle Scholar
  10. 10.
    Peretz D, Williamson RA, Legname G, Matsunaga Y, Vergara J, Burton DR, DeArmond SJ, Prusiner SB et al (2002) A change in the conformation of prions accompanies the emergence of a new prion strain. Neuron 34(6):921–932CrossRefGoogle Scholar
  11. 11.
    Cescatti M, Saverioni D, Capellari S, Tagliavini F, Kitamoto T, Ironside J, Giese A, Parchi P (2016) Analysis of conformational stability of abnormal prion protein aggregates across the spectrum of Creutzfeldt-Jakob disease prions. J Virol 90(14):6244–6254CrossRefGoogle Scholar
  12. 12.
    Fraser H (1993) Diversity in the neuropathology of scrapie-like diseases in animals. Br Med Bull 49(4):792–809CrossRefGoogle Scholar
  13. 13.
    DeArmond SJ, Sanchez H, Yehiely F, Qiu Y, Ninchak-Casey A, Daggett V, Camerino AP, Cayetano J et al (1997) Selective neuronal targeting in prion disease. Neuron 19(6):1337–1348CrossRefGoogle Scholar
  14. 14.
    Budka H (2003) Neuropathology of prion diseases. Br Med Bull 66:121–130CrossRefGoogle Scholar
  15. 15.
    Fraser H, Dickinson AG (1968) The sequential development of the brain lesion of scrapie in three strains of mice. J Comp Pathol 78(3):301–311CrossRefGoogle Scholar
  16. 16.
    Kovacs GG, Budka H (2008) Prion diseases: from protein to cell pathology. Am J Pathol 172(3):555–565CrossRefGoogle Scholar
  17. 17.
    Dickinson AG, Meikle VM (1971) Host-genotype and agent effects in scrapie incubation: change in allelic interaction with different strains of agent. Mol Gen Genet 112(1):73–79CrossRefGoogle Scholar
  18. 18.
    Hughes D, Halliday M (2017) What is our current understanding of PrP(Sc)-associated neurotoxicity and its molecular underpinnings? Pathogens 6(4)Google Scholar
  19. 19.
    Vilette D, Courte J, Peyrin JM, Coudert L, Schaeffer L, Andreoletti O, Leblanc P (2018) Cellular mechanisms responsible for cell-to-cell spreading of prions. Cell Mol Life SciGoogle Scholar
  20. 20.
    Gousset K, Zurzolo C (2009) Tunnelling nanotubes: a highway for prion spreading? Prion 3(2):94–98CrossRefGoogle Scholar
  21. 21.
    Nussbaum-Krammer CI, Park KW, Li L, Melki R, Morimoto RI (2013) Spreading of a prion domain from cell-to-cell by vesicular transport in Caenorhabditis elegans. PLoS Genet 9(3):e1003351CrossRefGoogle Scholar
  22. 22.
    Costanzo M, Zurzolo C (2013) The cell biology of prion-like spread of protein aggregates: mechanisms and implication in neurodegeneration. Biochem J 452(1):1–17CrossRefGoogle Scholar
  23. 23.
    Hartmann A, Muth C, Dabrowski O, Krasemann S, Glatzel M (2017) Exosomes and the prion protein: more than one truth. Front Neurosci 11:194CrossRefGoogle Scholar
  24. 24.
    Emamzadeh FN (2016) Alpha-synuclein structure, functions, and interactions. J Res Med Sci 21:29CrossRefGoogle Scholar
  25. 25.
    Yu S, Li X, Liu G, Han J, Zhang C, Li Y, Xu S, Liu C et al (2007) Extensive nuclear localization of alpha-synuclein in normal rat brain neurons revealed by a novel monoclonal antibody. Neuroscience 145(2):539–555CrossRefGoogle Scholar
  26. 26.
    Totterdell S, Meredith GE (2005) Localization of alpha-synuclein to identified fibers and synapses in the normal mouse brain. Neuroscience 135(3):907–913CrossRefGoogle Scholar
  27. 27.
    Ostrerova N, Petrucelli L, Farrer M, Mehta N, Choi P, Hardy J, Wolozin B (1999) Alpha-synuclein shares physical and functional homology with 14-3-3 proteins. J Neurosci 19(14):5782–5791CrossRefGoogle Scholar
  28. 28.
    Chen RH, Wislet-Gendebien S, Samuel F, Visanji NP, Zhang G, Marsilio D, Langman T, Fraser PE et al (2013) Alpha-synuclein membrane association is regulated by the Rab3a recycling machinery and presynaptic activity. J Biol Chem 288(11):7438–7449CrossRefGoogle Scholar
  29. 29.
    Peng X, Tehranian R, Dietrich P, Stefanis L, Perez RG (2005) Alpha-synuclein activation of protein phosphatase 2A reduces tyrosine hydroxylase phosphorylation in dopaminergic cells. J Cell Sci 118(Pt 15):3523–3530CrossRefGoogle Scholar
  30. 30.
    Burre J, Sharma M, Tsetsenis T, Buchman V, Etherton MR, Sudhof TC (2010) Alpha-synuclein promotes SNARE-complex assembly in vivo and in vitro. Science 329(5999):1663–1667CrossRefGoogle Scholar
  31. 31.
    Pranke IM, Morello V, Bigay J, Gibson K, Verbavatz JM, Antonny B, Jackson CL (2011) Alpha-synuclein and ALPS motifs are membrane curvature sensors whose contrasting chemistry mediates selective vesicle binding. J Cell Biol 194(1):89–103CrossRefGoogle Scholar
  32. 32.
    Witt SN (2010) Hsp70 molecular chaperones and Parkinson's disease. Biopolymers 93(3):218–228CrossRefGoogle Scholar
  33. 33.
    Witt SN (2013) Molecular chaperones, alpha-synuclein, and neurodegeneration. Mol Neurobiol 47(2):552–560CrossRefGoogle Scholar
  34. 34.
    Rekas A, Ahn KJ, Kim J, Carver JA (2012) The chaperone activity of alpha-synuclein: utilizing deletion mutants to map its interaction with target proteins. Proteins 80(5):1316–1325CrossRefGoogle Scholar
  35. 35.
    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):3095–3099CrossRefGoogle Scholar
  36. 36.
    Zhu M, Qin ZJ, Hu D, Munishkina LA, Fink AL (2006) Alpha-synuclein can function as an antioxidant preventing oxidation of unsaturated lipid in vesicles. Biochemistry 45(26):8135–8142CrossRefGoogle Scholar
  37. 37.
    Liu X, Lee YJ, Liou LC, Ren Q, Zhang Z, Wang S, Witt SN (2011) Alpha-synuclein functions in the nucleus to protect against hydroxyurea-induced replication stress in yeast. Hum Mol Genet 20(17):3401–3414CrossRefGoogle Scholar
  38. 38.
    Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388(6645):839–840CrossRefGoogle Scholar
  39. 39.
    Spillantini MG, Crowther RA, Jakes R, Cairns NJ, Lantos PL, Goedert M (1998) Filamentous alpha-synuclein inclusions link multiple system atrophy with Parkinson’s disease and dementia with Lewy bodies. Neurosci Lett 251(3):205–208CrossRefGoogle Scholar
  40. 40.
    Aulic S, Le TT, Moda F, Abounit S, Corvaglia S, Casalis L, Gustincich S, Zurzolo C et al (2014) Defined alpha-synuclein prion-like molecular assemblies spreading in cell culture. BMC Neurosci 15:69CrossRefGoogle Scholar
  41. 41.
    Luk KC, Kehm V, Carroll J, Zhang B, O’Brien P, Trojanowski JQ, Lee VMY (2012) Pathological alpha-Synuclein transmission initiates Parkinson-like neurodegeneration in nontransgenic mice. Science 338(6109):949–953CrossRefGoogle Scholar
  42. 42.
    Watts JC, Giles K, Oehler A, Middleton L, Dexter DT, Gentleman SM, DeArmond SJ, Prusiner SB (2013) Transmission of multiple system atrophy prions to transgenic mice. Proc Natl Acad Sci U S A 110(48):19555–19560CrossRefGoogle Scholar
  43. 43.
    Masuda-Suzukake M, Nonaka T, Hosokawa M, Oikawa T, Arai T, Akiyama H, Mann DM, Hasegawa M (2013) Prion-like spreading of pathological alpha-synuclein in brain. Brain 136(Pt 4):1128–1138CrossRefGoogle Scholar
  44. 44.
    Tarutani A, Arai T, Murayama S, Hisanaga SI, Hasegawa M (2018) Potent prion-like behaviors of pathogenic alpha-synuclein and evaluation of inactivation methods. Acta Neuropathol Commun 6(1):29CrossRefGoogle Scholar
  45. 45.
    Aulic S, Masperone L, Narkiewicz J, Isopi E, Bistaffa E, Ambrosetti E, Pastore B, De Cecco E et al (2017) Alpha-synuclein amyloids hijack prion protein to gain cell entry, facilitate cell-to-cell spreading and block prion replication. Sci Rep 7(1):10050CrossRefGoogle Scholar
  46. 46.
    Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Garcia-Aznar JM, Vila M, Samitier J et al (2018) Involvement of cellular prion protein in alpha-synuclein transport in neurons. Mol Neurobiol 55(3):1847–1860CrossRefGoogle Scholar
  47. 47.
    Katorcha E, Makarava N, Lee YJ, Lindberg I, Monteiro MJ, Kovacs GG, Baskakov IV (2017) Cross-seeding of prions by aggregated alpha-synuclein leads to transmissible spongiform encephalopathy. PLoS Pathog 13(8):e1006563CrossRefGoogle Scholar
  48. 48.
    Bartz JC, Bessen RA, McKenzie D, Marsh RF, Aiken JM (2000) Adaptation and selection of prion protein strain conformations following interspecies transmission of transmissible mink encephalopathy. J Virol 74(12):5542–5547CrossRefGoogle Scholar
  49. 49.
    Bistaffa E, Moda F, Virgilio T, Campagnani I, De Luca CMG, Rossi M, Salzano G, Giaccone G et al (2018) Synthetic prion selection and adaptation. Mol NeurobiolGoogle Scholar
  50. 50.
    Baskakov IV (2014) The many shades of prion strain adaptation. Prion 8(2)CrossRefGoogle Scholar
  51. 51.
    Katorcha E, Gonzalez-Montalban N, Makarava N, Kovacs GG, Baskakov IV (2018) Prion replication environment defines the fate of prion strain adaptation. PLoS Pathog 14(6):e1007093CrossRefGoogle Scholar
  52. 52.
    Specht CG, Schoepfer R (2001) Deletion of the alpha-synuclein locus in a subpopulation of C57BL/6J inbred mice. BMC Neurosci 2:11CrossRefGoogle Scholar
  53. 53.
    Saborio GP, Permanne B, Soto C (2001) Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature 411(6839):810–813CrossRefGoogle Scholar
  54. 54.
    Cabin DE, Shimazu K, Murphy D, Cole NB, Gottschalk W, McIlwain KL, Orrison B, Chen A et al (2002) Synaptic vesicle depletion correlates with attenuated synaptic responses to prolonged repetitive stimulation in mice lacking alpha-synuclein. J Neurosci 22(20):8797–8807CrossRefGoogle Scholar
  55. 55.
    Dauer W, Kholodilov N, Vila M, Trillat AC, Goodchild R, Larsen KE, Staal R, Tieu K et al (2002) Resistance of alpha -synuclein null mice to the parkinsonian neurotoxin MPTP. Proc Natl Acad Sci U S A 99(22):14524–14529CrossRefGoogle Scholar
  56. 56.
    Chandra S, Gallardo G, Fernandez-Chacon R, Schluter OM, Sudhof TC (2005) Alpha-synuclein cooperates with CSPalpha in preventing neurodegeneration. Cell 123(3):383–396CrossRefGoogle Scholar
  57. 57.
    Lindberg I, Shorter J, Wiseman RL, Chiti F, Dickey CA, McLean PJ (2015) Chaperones in neurodegeneration. J Neurosci 35(41):13853–13859CrossRefGoogle Scholar
  58. 58.
    St Martin JL, Klucken J, Outeiro TF, Nguyen P, Keller-McGandy C, Cantuti-Castelvetri I, Grammatopoulos TN, Standaert DG et al (2007) Dopaminergic neuron loss and up-regulation of chaperone protein mRNA induced by targeted over-expression of alpha-synuclein in mouse substantia nigra. J Neurochem 100(6):1449–1457PubMedGoogle Scholar
  59. 59.
    Nakamura S, Ono F, Hamano M, Odagiri K, Kubo M, Komatsuzaki K, Terao K, Shinagawa M et al (2000) Immunohistochemical detection of apolipoprotein E within prion-associated lesions in squirrel monkey brains. Acta Neuropathol 100(4):365–370CrossRefGoogle Scholar
  60. 60.
    Hochstrasser DF, Frutiger S, Wilkins MR, Hughes G, Sanchez JC (1997) Elevation of apolipoprotein E in the CSF of cattle affected by BSE. FEBS Lett 416(2):161–163CrossRefGoogle Scholar
  61. 61.
    Skinner PJ, Abbassi H, Chesebro B, Race RE, Reilly C, Haase AT (2006) Gene expression alterations in brains of mice infected with three strains of scrapie. BMC Genomics 7:114CrossRefGoogle Scholar
  62. 62.
    Van Everbroeck B, Croes EA, Pals P, Dermaut B, Jansen G, van Duijn CM, Cruts M, Van Broeckhoven C et al (2001) Influence of the prion protein and the apolipoprotein E genotype on the Creutzfeldt-Jakob disease phenotype. Neurosci Lett 313(1–2):69–72CrossRefGoogle Scholar
  63. 63.
    Baumann MH, Kallijarvi J, Lankinen H, Soto C, Haltia M (2000) Apolipoprotein E includes a binding site which is recognized by several amyloidogenic polypeptides. Biochem J 349(Pt 1):77–84CrossRefGoogle Scholar
  64. 64.
    Moore RA, Timmes AG, Wilmarth PA, Safronetz D, Priola SA (2011) Identification and removal of proteins that co-purify with infectious prion protein improves the analysis of its secondary structure. Proteomics 11(19):3853–3865CrossRefGoogle Scholar
  65. 65.
    Altmeppen HC, Puig B, Dohler F, Thurm DK, Falker C, Krasemann S, Glatzel M Proteolytic processing of the prion protein in health and disease. Am J Neurodegener Dis 2012(1, 1):15–31Google Scholar
  66. 66.
    Iida T, Doh-ura K, Kawashima T, Abe H, Iwaki T (2001) An atypical case of sporadic Creutzfeldt-Jakob disease with Parkinson's disease. Neuropathology 21(4):294–297CrossRefGoogle Scholar
  67. 67.
    Haik S, Privat N, Adjou KT, Sazdovitch V, Dormont D, Duyckaerts C, Hauw JJ (2002) Alpha-synuclein-immunoreactive deposits in human and animal prion diseases. Acta Neuropathol 103(5):516–520CrossRefGoogle Scholar
  68. 68.
    Adjou KT, Allix S, Ouidja MO, Backer S, Couquet C, Cornuejols MJ, Deslys JP, Brugere H et al (2007) Alpha-synuclein accumulates in the brain of scrapie-affected sheep and goats. J Comp Pathol 137(1):78–81CrossRefGoogle Scholar
  69. 69.
    La Vitola P, Beeg M, Balducci C, Santamaria G, Restelli E, Colombo L, Caldinelli L, Pollegioni L et al (2019) Cellular prion protein neither binds to alpha-synuclein oligomers nor mediates their detrimental effects. Brain 142(2):249–254CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Edoardo Bistaffa
    • 1
  • Martina Rossi
    • 2
  • Chiara Maria Giulia De Luca
    • 1
    • 2
  • Federico Cazzaniga
    • 1
  • Olga Carletta
    • 1
  • Ilaria Campagnani
    • 1
  • Fabrizio Tagliavini
    • 1
  • Giuseppe Legname
    • 2
  • Giorgio Giaccone
    • 1
  • Fabio Moda
    • 1
    Email author
  1. 1.Unit of Neuropathology and Neurology 5Fondazione IRCCS Istituto Neurologico Carlo BestaMilanItaly
  2. 2.Department of Neuroscience, Laboratory of Prion BiologyScuola Internazionale Superiore di Studi Avanzati (SISSA)TriesteItaly

Personalised recommendations