Change search
ReferencesLink to record
Permanent link

Direct link
Antibodies against alpha-synuclein reduce oligomerization in living cells
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics. (Lannfelt, Molekylär geriatrik/ Rudbecklaboratoriet)
Cell and Molecular Neuroscience Unit, Instituto de Medicina Molecular, Lisboa.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics.
BioArctic Neuroscience AB.
Show others and affiliations
2011 (English)In: PLoS ONE, ISSN 1932-6203, Vol. 6, no 10, e27230- p.Article in journal (Refereed) Published
Abstract [en]

Recent research implicates soluble aggregated forms of α-synuclein as neurotoxic species with a central role in the pathogenesis of Parkinson’s disease and related disorders. The pathway by which α-synuclein aggregates is believed to follow a step-wise pattern, in which dimers and smaller oligomers are initially formed. Here, we used H4 neuroglioma cells expressing α-synuclein fused to hemi:GFP to study the effects of α-synuclein monoclonal antibodies on the early stages of aggregation, as quantified by Bimolecular Fluorescence Complementation assay. Widefield and confocal microscopy revealed that cells treated for 48 h with monoclonal antibodies internalized antibodies to various degrees. Oligomer-selective and C-terminal specific α-synuclein antibodies reduced the extent of α-synuclein dimerization/oligomerization, as indicated by decreased GFP fluorescence signal. Furthermore, ELISA measurements on lysates and conditioned media from antibody treated cells displayed lower α-synuclein levels compared to untreated cells, suggesting increased protein turnover. Taken together, our results propose that extracellular administration of monoclonal antibodies can modify or inhibit early steps in the aggregation process of α-synuclein, thus providing further support for passive immunization against diseases with α-synuclein pathology.

Place, publisher, year, edition, pages
2011. Vol. 6, no 10, e27230- p.
Keyword [en]
Alpha-synuclein; Parkinson’s disease; Lewy bodies; Aggregation; Bimolecular Fluorescence Complementation; Monoclonal antibodies; Immunotherapy
National Category
Neurosciences Cell and Molecular Biology
Research subject
Neuroscience; Geriatrics
URN: urn:nbn:se:uu:diva-160097DOI: 10.1371/journal.pone.0027230ISI: 000296916000059PubMedID: 22073131OAI: diva2:450411
Available from: 2011-10-20 Created: 2011-10-16 Last updated: 2013-01-09Bibliographically approved
In thesis
1. Characterization of α-synuclein oligomers: Implications for Lewy Body Disorders
Open this publication in new window or tab >>Characterization of α-synuclein oligomers: Implications for Lewy Body Disorders
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Parkinson’s disease, dementia with Lewy bodies and multiple system atrophy are disorders featuring accumulation of Lewy bodies in brain. The main component of these large insoluble intracellular inclusions is the presynaptic protein alpha-synuclein (α-synuclein). It is generally believed that α-synuclein monomers adopt an abnormal conformation that favors the formation of soluble oligomers or protofibrils and, eventually, insoluble fibrils depositing as Lewy bodies. Notably, the intermediately sized oligomers/protofibrils seem to have particular neurotoxic effects. Several factors may influence the formation of α-synuclein oligomers/protofibrils, e.g. the reactive aldehydes 4-hydroxy-2-nonenal (HNE) and 4-oxo-2-nonenal (ONE) formed during oxidative stress.

The overall aims of this thesis were to investigate biophysical and biochemical properties of in vitro generated α-synuclein oligomers, characterize their functional effects on cell and animal disease models as well as to explore whether their formation could be prevented in a cell culture model for oligomerization. 

Here, it was found that α-synuclein rapidly formed oligomers after incubation with both ONE and HNE. The resulting oligomers were stable and did not continue to form insoluble fibrils. By comparing HNE- and ONE induced α-synuclein oligomers biochemically they were both found to exhibit extensive β-beta sheet structure and had a molecular size of ~2000 kDa. However, they differed in morphology; the ONE induced α-synuclein oligomers described round amorphous species whereas the HNE induced α-synuclein oligomers appeared as elongated protofibril-like structures. Both these oligomers were cell internalized to varying degrees and induced toxicity in neuroblastoma cells.

In addition, the ONE induced α-synuclein oligomers seemed to initiate aggregation of monomeric α-synuclein in vitro, but failed to do so in vivo.

Finally, treatment of α-synuclein overexpressing cells with monoclonal antibodies specific for α-synuclein significantly reduced aggregation and lowered levels of the protein, suggesting increased turnover in these cells. 

To conclude, this thesis has characterized different oligomeric α-synuclein species, which may have properties similar to soluble species central to the pathogenesis of Parkinson’s disease and other disorders with α-synuclein pathology. For therapeutic strategies it is important to selectively target such harmful protein species and avoid interaction with other forms of α-synuclein, which may have vital physiological cellular functions.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2011. 70 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 717
Parkinson’s disease; Alpha-synuclein; Lewy bodies; Oligomers; Reactive aldehydes; monoclonal antibody
National Category
Neurosciences Geriatrics
Research subject
Medical Science
urn:nbn:se:uu:diva-160102 (URN)978-91-554-8198-8 (ISBN)
Public defence
2011-12-02, Rudbecksalen, Rudbecklaboratoriet, Dag Hammarskjöldsväg 20, Uppsala, 13:15 (English)
Available from: 2011-11-11 Created: 2011-10-16 Last updated: 2011-11-23Bibliographically approved

Open Access in DiVA

fulltext(362 kB)160 downloads
File information
File name FULLTEXT01.pdfFile size 362 kBChecksum SHA-512
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Näsström, ThomasLannfelt, LarsBergström, JoakimIngelsson, Martin
By organisation
In the same journal
NeurosciencesCell and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 160 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 215 hits
ReferencesLink to record
Permanent link

Direct link