Formation of toxic oligomeric α-synuclein species in living cells

Tiago Fleming Outeiro, Preeti Putcha, Julie E. Tetzlaff, Robert Spoelgen, Mirjam Koker, Filipe Carvalho, Bradley T. Hyman, Pamela J. McLean

Research output: Contribution to journalArticle

280 Scopus citations

Abstract

Background: Misfolding, oligomerization, and fibrillization of α-synuclein are thought to be central events in the onset and progression of Parkinson's desease (PD) and related disorders. Although fibrillar α-synuclein is a major components of Lewy bodies (LBs), recent data implicate prefibrillar, oligomeric intermediates as the toxic species. However, to date, oligomeric species have not-been identified in living cells. Methodology/Principal Findings: Here we used bimolecular flourescence complementation (BiFC) to directly visualize α-synuclein oligomerization in living cells, allowing us to study the initial events leading to α-synuclein oligomerization, the precursor to aggregate formation. This novel assay provides us with a tool with which to investigate how manipulations affecting α-synuclein aggregation affect the process over time. Stabilization of α-synuclein oligomers via BiFC results in increased cytoxixity, which can be rescued by Hsp70 in a process that reduces the formation of α-synuclein oligomers. Introduction of PD-assisted mutations in α-synuclein did not affect oligomer formation but the biochemical properties of the mutant α-synuclein oligomers differ from those of wild type α-synuclein. Conclusion/Significance: This novel application of the BiFC assay to the study of the molecular basis of neurodegenerative disroders enabled the direct vissualization of α-synuclein oligomeric species in living cells and its mudulation by Hsp70, constituting a novel important tool in the search for the synucleinopathies.

Original languageEnglish (US)
Article numbere1867
JournalPloS one
Volume3
Issue number4
DOIs
StatePublished - Apr 2 2008

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Outeiro, T. F., Putcha, P., Tetzlaff, J. E., Spoelgen, R., Koker, M., Carvalho, F., Hyman, B. T., & McLean, P. J. (2008). Formation of toxic oligomeric α-synuclein species in living cells. PloS one, 3(4), [e1867]. https://doi.org/10.1371/journal.pone.0001867