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  Iron-mediated aggregation and toxicity in a novel neuronal cell culture model with inducible alpha-synuclein expression.

Bartels, M., Weckbecker, D., Kuhn, P. H., Ryazanov, S., Leonov, A., Griesinger, C., et al. (2019). Iron-mediated aggregation and toxicity in a novel neuronal cell culture model with inducible alpha-synuclein expression. Scientific Reports, 9: 9100. doi:10.1038/s41598-019-45298-6.

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Bartels, M., Autor
Weckbecker, D., Autor
Kuhn, P. H., Autor
Ryazanov, S.1, Autor           
Leonov, A.2, Autor           
Griesinger, C.2, Autor                 
Lichtenthaler, S. F., Autor
Bötzel, K., Autor
Giese, A., Autor
Affiliations:
1Department of NMR-based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              
2Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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 Zusammenfassung: Parkinson's disease (PD) represents an increasing problem in society. The oligomerization of alpha-synuclein (αSyn) is a suggested key event in its pathogenesis, yet the pathological modes of action remain to be fully elucidated. To identify potential disease-modifying therapeutics and to study αSyn-mediated toxic mechanisms, we established cell lines with inducible overexpression of different αSyn constructs: αSyn, αSyn coupled to the fluorescence protein Venus (αSyn-Venus), and αSyn coupled to the N-terminal or C-terminal part of Venus (V1S and SV2, respectively) for a bimolecular fluorescence complementation assay (BiFC). Inducibility was achieved by applying modified GAL4-UAS or Cre-loxP systems and addition of tebufenozide or 4-OH-tamoxifen, respectively. Expression constructs were stably integrated into the host genome of H4 neuroglioma cells by lentiviral transduction. We here demonstrate a detailed investigation of the expression characteristics of inducible H4 cells showing low background expression and high inducibility. We observed increased protein load and aggregation of αSyn upon incubation with DMSO and FeCl3 along with an increase in cytotoxicity. In summary, we present a system for the creation of inducibly αSyn-overexpressing cell lines holding high potential for the screening for modulators of αSyn aggregation and αSyn-mediated toxicity.

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Sprache(n): eng - English
 Datum: 2019-06-24
 Publikationsstatus: Online veröffentlicht
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 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1038/s41598-019-45298-6
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Titel: Scientific Reports
Genre der Quelle: Zeitschrift
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Seiten: 13 Band / Heft: 9 Artikelnummer: 9100 Start- / Endseite: - Identifikator: -