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Molecular Neurobiology

, Volume 56, Issue 4, pp 2978–2989 | Cite as

Synthetic Prion Selection and Adaptation

  • Edoardo Bistaffa
  • Fabio Moda
  • Tommaso Virgilio
  • Ilaria Campagnani
  • Chiara Maria Giulia De Luca
  • Martina Rossi
  • Giulia Salzano
  • Giorgio Giaccone
  • Fabrizio Tagliavini
  • Giuseppe LegnameEmail author
Article

Abstract

Prion pathologies are characterized by the conformational conversion of the cellular prion protein (PrPC) into a pathological infectious isoform, known as PrPSc. The latter acquires different abnormal conformations, which are associated with specific pathological phenotypes. Recent evidence suggests that prions adapt their conformation to changes in the context of replication. This phenomenon is known as either prion selection or adaptation, where distinct conformations of PrPSc with higher propensity to propagate in the new environment prevail over the others. Here, we show that a synthetically generated prion isolate, previously subjected to protein misfolding cyclic amplification (PMCA) and then injected in animals, is able to change its biochemical and biophysical properties according to the context of replication. In particular, in second transmission passage in vivo, two different prion isolates were found: one characterized by a predominance of the monoglycosylated band (PrPSc-M) and the other characterized by a predominance of the diglycosylated one (PrPSc-D). Neuropathological, biochemical, and biophysical assays confirmed that these PrPSc possess distinctive characteristics. Finally, PMCA analysis of PrPSc-M and PrPSc-D generated PrPSc (PrPSc-PMCA) whose biophysical properties were different from those of both inocula, suggesting that PMCA selectively amplified a third PrPSc isolate. Taken together, these results indicate that the context of replication plays a pivotal role in either prion selection or adaptation. By exploiting the ability of PMCA to mimic the process of prion replication in vitro, it might be possible to assess how changes in the replication environment influence the phenomenon of prion selection and adaptation.

Keywords

Synthetic prion Prion Adaptation Selection PMCA RT-QuIC 

Notes

Acknowledgements

The authors wish to thank Associazione Italiana Encefalopatie da Prioni (A.I.En.P.).

Authors Contributions

EB, FM, and GL designed the experiments and EB, TV, IC, CMGDL, MR, and GS performed the practical work. TV and IC performed the animal inoculations. EB, FM, GG, FT, and GL wrote and revised the manuscript. All the authors read and approved the final manuscript.

Funding

This work was supported/partially supported by the Italian Ministry of Health (GR-2013-02355724 and RC) to FM, the Italian Ministry of Health to FT, and the International School for Advanced Studies (SISSA) intramural funding to GL.

Compliance with Ethical Standards

The study, including its Ethics aspects, was approved by the Italian Ministry of Health (Permit Number, NP-02-14).

Conflict of Interest

The authors declare that there are no conflicts of interest.

Supplementary material

12035_2018_1279_MOESM1_ESM.jpg (211 kb)
ESM 1 Biochemical characterization of PrPSc in eyes and spleen of animals with PrPSc-M and PrPSc-D. (JPG 210 kb)
12035_2018_1279_MOESM2_ESM.jpg (189 kb)
ESM 2 PMCA analysis of PrPSc-M and PrPSc-D spleen-derived PrPSc. (JPG 188 kb)
12035_2018_1279_MOESM3_ESM.jpg (352 kb)
ESM 3 ThS staining of frontal cortex of mice with PrPSc-M and PrPSc-D. (JPG 352 kb)
12035_2018_1279_MOESM4_ESM.jpg (769 kb)
ESM 4 PMCA amplification of serial dilutions of PrPSc-M and PrPSc-D. PrPSc-M and PrPSc-D were serially diluted in normal mouse brain homogenates and subjected to 2 serial rounds of amplification by means of PMCA. All amplified samples showed a PrPSc characterized by a prevalence of the diglycosylated isoform of the protein. (JPG 768 kb)
12035_2018_1279_MOESM5_ESM.jpg (608 kb)
ESM 5 RT-QuIC assay of PrPSc-M and PrPSc-D isolates. (A) Representative kinetic curves of recombinant mouse PrP (recPrP) seeded with PrPSc-M and PrPSc-D and PrPSc-PMCA. (B) Analysis of the slope of PrPSc-M and PrPSc-D and PrPSc-PMCA aggregation kinetic curves. (* p<0.05, ** p<0.01) (C) Analysis of the slope of RML and RML-PMCA aggregation kinetic curves. (JPG 608 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Edoardo Bistaffa
    • 1
    • 2
  • Fabio Moda
    • 2
  • Tommaso Virgilio
    • 2
    • 3
  • Ilaria Campagnani
    • 2
  • Chiara Maria Giulia De Luca
    • 2
  • Martina Rossi
    • 1
  • Giulia Salzano
    • 1
  • Giorgio Giaccone
    • 2
  • Fabrizio Tagliavini
    • 2
  • Giuseppe Legname
    • 1
    • 4
    Email author
  1. 1.Laboratory of Prion Biology, Department of NeuroscienceScuola Internazionale Superiore di Studi Avanzati (SISSA)TriesteItaly
  2. 2.Unit of Neuropathology and Neurology 5Fondazione IRCCS Istituto Neurologico Carlo BestaMilanItaly
  3. 3.Institute for Research in BiomedicineUniversità della Svizzera ItalianaBellinzonaSwitzerland
  4. 4.ELETTRA LaboratorySincrotrone Trieste S.C.p.ATriesteItaly

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