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.; de Pomerai, David I.

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.

Citation

Bodhicharla, R., Nagarajan, A., Winter, J., Adenle, A., Nazir, A., Brady, D., Vere, K., Richens, J., O'Shea, P., Bell, D. R., & de Pomerai, D. I. (2012). Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains. CNS and Neurological Disorders - Drug Targets, 11(8), https://doi.org/10.2174/1871527311211080005

• Journal Article Type: Article
• Publication Date: Jan 1, 2012
• Deposit Date: Apr 16, 2014
• Publicly Available Date: Apr 16, 2014
• Journal: CNS and Neurological Disorders - Drug Targets
• Print ISSN: 1871-5273
• Electronic ISSN: 1996-3181
• Publisher: Bentham Science Publishers
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Issue: 8
• DOI: https://doi.org/10.2174/1871527311211080005
• Keywords: Caenorhabditis elegans, CFP and YFP reporters, FRET, protein aggregation, α-synuclein, transgenic strains
• Public URL: https://nottingham-repository.worktribe.com/output/1008273
• Publisher URL: http://europepmc.org/articles/PMC3744922/reload=0;jsessionid=JPd76GgNT9LPI7BNrhZt.12