Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains

Bodhicharla, Rakesh, Nagarajan, Archana, Winter, Jody, Adenle, Ademola, Nazir, Aamir, Brady, Declan, Vere, Kelly, Richens, Jo, O'Shea, Paul, Bell, David R. and de Pomerai, David I. (2012) Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains. CNS and Neurological Disorders - Drug Targets, 11 (8). pp. 965-975. ISSN 1871-5273

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Abstract

The neural protein α-synuclein aggregates both in vivo and in vitro to form insoluble fibrils that are involved in

Parkinson’s disease pathogenesis. We have generated α-synuclein/fluorescent-protein fusion constructs overexpressed in muscle cells of the nematode, Caenorhabdtis elegans. Green Fluorescent Protein (GFP) variants, Cerulean (C) or Venus (V), were fused to the C-terminus of human α-synuclein (S); the resultant fusion genes were designated SV and SC, plus a CV fusion as well as S, C and V singly. The aggregation behavior of the purified fusion proteins (expressed in E. coli) will be

described elsewhere. These constructs were fused to a C. elegans unc-54 myosin promoter, and integrated transgenic lines generated by microinjection, gamma-irradiation, and outcrossing of fluorescent progeny. All transgenic lines expressing α-synuclein showed significant reductions (p < 0.05) in lifespan, motility and pharyngeal pumping, as compared to wildtype worms or lines expressing CFP and/or YFP only. We showed that CFP and YFP labels colocalised in granular inclusions throughout the body wall in transgenic lines expressing both SC and SV fusions (SC+SV), whereas SV+C worms.

Item Type: Article
RIS ID: https://nottingham-repository.worktribe.com/output/1008273
Keywords: Caenorhabditis elegans, CFP and YFP reporters, FRET, protein aggregation, α-synuclein, transgenic strains
Schools/Departments: University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Life Sciences > School of Biology
Identification Number: https://doi.org/10.2174/1871527311211080005
Depositing User: Liu, Mr Zhenxing
Date Deposited: 16 Apr 2014 12:52
Last Modified: 04 May 2020 20:22
URI: https://eprints.nottingham.ac.uk/id/eprint/2591

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