Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains

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 Fluore...

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Main Authors: 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.
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Published: Bentham Science Publishers 2012
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Online Access:https://eprints.nottingham.ac.uk/2591/
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author 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.
author_facet 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.
author_sort Bodhicharla, Rakesh
building Nottingham Research Data Repository
collection Online Access
description 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.
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spelling nottingham-25912020-05-04T20:22:01Z https://eprints.nottingham.ac.uk/2591/ 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. 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. Bentham Science Publishers 2012 Article PeerReviewed 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 Caenorhabditis elegans CFP and YFP reporters FRET protein aggregation α-synuclein transgenic strains http://europepmc.org/articles/PMC3744922/reload=0;jsessionid=JPd76GgNT9LPI7BNrhZt.12 doi:10.2174/1871527311211080005 doi:10.2174/1871527311211080005
spellingShingle Caenorhabditis elegans
CFP and YFP reporters
FRET
protein aggregation
α-synuclein
transgenic 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.
Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title_full Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title_fullStr Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title_full_unstemmed Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title_short Effects of α-synuclein overexpression in transgenic Caenorhabditis elegans strains
title_sort effects of α-synuclein overexpression in transgenic caenorhabditis elegans strains
topic Caenorhabditis elegans
CFP and YFP reporters
FRET
protein aggregation
α-synuclein
transgenic strains
url https://eprints.nottingham.ac.uk/2591/
https://eprints.nottingham.ac.uk/2591/
https://eprints.nottingham.ac.uk/2591/