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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.

Authors

Rakesh Bodhicharla

Archana Nagarajan

Jody Winter

Ademola Adenle

Aamir Nazir

Declan Brady

Kelly Vere

Jo Richens

Paul O'Shea

David R. Bell

David I. de Pomerai david.depomerai@nottingham.ac.uk



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.

Journal Article Type Article
Publication Date Jan 1, 2012
Journal CNS and Neurological Disorders - Drug Targets
Print ISSN 1871-5273
Electronic ISSN 1871-5273
Publisher Bentham Open
Peer Reviewed Peer Reviewed
Volume 11
Issue 8
APA6 Citation Bodhicharla, R., Nagarajan, A., Winter, J., Adenle, A., Nazir, A., Brady, D., …de Pomerai, D. I. (2012). Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains. CNS and Neurological Disorders - Drug Targets, 11(8), doi:10.2174/1871527311211080005
DOI https://doi.org/10.2174/1871527311211080005
Keywords Caenorhabditis elegans, CFP and YFP reporters, FRET, protein aggregation, α-synuclein, transgenic strains
Publisher URL http://europepmc.org/articles/PMC3744922/reload=0;jsessionid=JPd76GgNT9LPI7BNrhZt.12
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0



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