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Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease

Huntington's disease (HD) is a late-onset neurodegenerative disorder that is characterized neuropathologically by the presence of neuropil aggregates and nuclear inclusions. However, the profile of aggregate structures that are present in the brains of HD patients or of HD mouse models and the...

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Autores principales: Sathasivam, Kirupa, Lane, Amin, Legleiter, Justin, Warley, Alice, Woodman, Ben, Finkbeiner, Steve, Paganetti, Paolo, Muchowski, Paul J., Wilson, Stuart, Bates, Gillian P.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2792149/
https://www.ncbi.nlm.nih.gov/pubmed/19825844
http://dx.doi.org/10.1093/hmg/ddp467
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author Sathasivam, Kirupa
Lane, Amin
Legleiter, Justin
Warley, Alice
Woodman, Ben
Finkbeiner, Steve
Paganetti, Paolo
Muchowski, Paul J.
Wilson, Stuart
Bates, Gillian P.
author_facet Sathasivam, Kirupa
Lane, Amin
Legleiter, Justin
Warley, Alice
Woodman, Ben
Finkbeiner, Steve
Paganetti, Paolo
Muchowski, Paul J.
Wilson, Stuart
Bates, Gillian P.
author_sort Sathasivam, Kirupa
collection PubMed
description Huntington's disease (HD) is a late-onset neurodegenerative disorder that is characterized neuropathologically by the presence of neuropil aggregates and nuclear inclusions. However, the profile of aggregate structures that are present in the brains of HD patients or of HD mouse models and the relative contribution of specific aggregate structures to disease pathogenesis is unknown. We have used the Seprion ligand to develop a highly sensitive enzyme-linked immunosorbent assay (ELISA)-based method for quantifying aggregated polyglutamine in tissues from HD mouse models. We used a combination of electron microscopy, atomic force microscopy (AFM) and sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) to investigate the aggregate structures isolated by the ligand. We found that the oligomeric, proto-fibrillar and fibrillar aggregates extracted from the brains of R6/2 and HdhQ150 knock-in mice were remarkably similar. Using AFM, we determined that the nanometre globular oligomers isolated from the brains of both mouse models have dimensions identical to those generated from recombinant huntingtin exon 1 proteins. Finally, antibodies that detect exon 1 Htt epitopes differentially recognize the ligand-captured material on SDS–PAGE gels. The Seprion-ligand ELISA provides an assay with good statistical power for use in preclinical pharmacodynamic therapeutic trials or to assess the effects of the genetic manipulation of potential therapeutic targets on aggregate load. This, together with the ability to identify a spectrum of aggregate species in HD mouse tissues, will contribute to our understanding of how these structures relate to the pathogenesis of HD and whether their formation can be manipulated for therapeutic benefit.
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spelling pubmed-27921492009-12-14 Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease Sathasivam, Kirupa Lane, Amin Legleiter, Justin Warley, Alice Woodman, Ben Finkbeiner, Steve Paganetti, Paolo Muchowski, Paul J. Wilson, Stuart Bates, Gillian P. Hum Mol Genet Articles Huntington's disease (HD) is a late-onset neurodegenerative disorder that is characterized neuropathologically by the presence of neuropil aggregates and nuclear inclusions. However, the profile of aggregate structures that are present in the brains of HD patients or of HD mouse models and the relative contribution of specific aggregate structures to disease pathogenesis is unknown. We have used the Seprion ligand to develop a highly sensitive enzyme-linked immunosorbent assay (ELISA)-based method for quantifying aggregated polyglutamine in tissues from HD mouse models. We used a combination of electron microscopy, atomic force microscopy (AFM) and sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) to investigate the aggregate structures isolated by the ligand. We found that the oligomeric, proto-fibrillar and fibrillar aggregates extracted from the brains of R6/2 and HdhQ150 knock-in mice were remarkably similar. Using AFM, we determined that the nanometre globular oligomers isolated from the brains of both mouse models have dimensions identical to those generated from recombinant huntingtin exon 1 proteins. Finally, antibodies that detect exon 1 Htt epitopes differentially recognize the ligand-captured material on SDS–PAGE gels. The Seprion-ligand ELISA provides an assay with good statistical power for use in preclinical pharmacodynamic therapeutic trials or to assess the effects of the genetic manipulation of potential therapeutic targets on aggregate load. This, together with the ability to identify a spectrum of aggregate species in HD mouse tissues, will contribute to our understanding of how these structures relate to the pathogenesis of HD and whether their formation can be manipulated for therapeutic benefit. Oxford University Press 2010-01-01 2009-10-12 /pmc/articles/PMC2792149/ /pubmed/19825844 http://dx.doi.org/10.1093/hmg/ddp467 Text en © The Author 2009. Published by Oxford University Press http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Sathasivam, Kirupa
Lane, Amin
Legleiter, Justin
Warley, Alice
Woodman, Ben
Finkbeiner, Steve
Paganetti, Paolo
Muchowski, Paul J.
Wilson, Stuart
Bates, Gillian P.
Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title_full Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title_fullStr Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title_full_unstemmed Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title_short Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease
title_sort identical oligomeric and fibrillar structures captured from the brains of r6/2 and knock-in mouse models of huntington's disease
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2792149/
https://www.ncbi.nlm.nih.gov/pubmed/19825844
http://dx.doi.org/10.1093/hmg/ddp467
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