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新型ウイルス 封鎖から1か月 先行きの見えない街 中国 武漢 (NHK)

新型コロナウイルスの感染が広がっている中国で状況が最も深刻な湖北省武漢では、事実上、街が封鎖される措置がとられてから23日で1か月になります。住民は外出を厳しく...

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

著者 Bartels M , Weckbecker D , Kuhn PH , Ryazanov S , Leonov A , Griesinger C , Lichtenthaler SF , Bötzel K , Giese A
Sci Rep.2019 Jun 24 ; 9(1):9100.
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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 FeCl 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.
PMID: 31235814 [PubMed - in process]
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