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

265 Citations (Scopus)

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
Externally publishedYes

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Synucleins
Poisons
Cells
assays
Oligomers
cells
cell aggregates
Oligomerization
mutation
mutants
Assays
oligomerization
Lewy Bodies
methodology

ASJC Scopus subject areas

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

Cite this

Outeiro, T. F., Putcha, P., Tetzlaff, J. E., Spoelgen, R., Koker, M., Carvalho, F., ... 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

Formation of toxic oligomeric α-synuclein species in living cells. / Outeiro, Tiago Fleming; Putcha, Preeti; Tetzlaff, Julie E.; Spoelgen, Robert; Koker, Mirjam; Carvalho, Filipe; Hyman, Bradley T.; McLean, Pamela J.

In: PLoS One, Vol. 3, No. 4, e1867, 02.04.2008.

Research output: Contribution to journalArticle

Outeiro, TF, Putcha, P, Tetzlaff, JE, Spoelgen, R, Koker, M, Carvalho, F, Hyman, BT & McLean, PJ 2008, 'Formation of toxic oligomeric α-synuclein species in living cells', PLoS One, vol. 3, no. 4, e1867. https://doi.org/10.1371/journal.pone.0001867
Outeiro TF, Putcha P, Tetzlaff JE, Spoelgen R, Koker M, Carvalho F et al. Formation of toxic oligomeric α-synuclein species in living cells. PLoS One. 2008 Apr 2;3(4). e1867. https://doi.org/10.1371/journal.pone.0001867
Outeiro, Tiago Fleming ; Putcha, Preeti ; Tetzlaff, Julie E. ; Spoelgen, Robert ; Koker, Mirjam ; Carvalho, Filipe ; Hyman, Bradley T. ; McLean, Pamela J. / Formation of toxic oligomeric α-synuclein species in living cells. In: PLoS One. 2008 ; Vol. 3, No. 4.
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