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Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates
Here we describe the use of an organotypic hippocampal slice model for studying α-synuclein aggregation and inter-neuronal spreading initiated by microinjection of pre-formed α-synuclein fibrils (PFFs). PFF injection at dentate gyrus (DG) templates the formation of endogenous α-synuclein aggregates...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6924077/ https://www.ncbi.nlm.nih.gov/pubmed/31856920 http://dx.doi.org/10.1186/s40478-019-0865-5 |
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author | Elfarrash, Sara Jensen, Nanna Møller Ferreira, Nelson Betzer, Cristine Thevathasan, Jervis Vermal Diekmann, Robin Adel, Mohamed Omar, Nisreen Mansour Boraie, Mohamed Z. Gad, Sabry Ries, Jonas Kirik, Deniz Nabavi, Sadegh Jensen, Poul Henning |
author_facet | Elfarrash, Sara Jensen, Nanna Møller Ferreira, Nelson Betzer, Cristine Thevathasan, Jervis Vermal Diekmann, Robin Adel, Mohamed Omar, Nisreen Mansour Boraie, Mohamed Z. Gad, Sabry Ries, Jonas Kirik, Deniz Nabavi, Sadegh Jensen, Poul Henning |
author_sort | Elfarrash, Sara |
collection | PubMed |
description | Here we describe the use of an organotypic hippocampal slice model for studying α-synuclein aggregation and inter-neuronal spreading initiated by microinjection of pre-formed α-synuclein fibrils (PFFs). PFF injection at dentate gyrus (DG) templates the formation of endogenous α-synuclein aggregates in axons and cell bodies of this region that spread to CA3 and CA1 regions. Aggregates are insoluble and phosphorylated at serine-129, recapitulating Lewy pathology features found in Parkinson’s disease and other synucleinopathies. The model was found to favor anterograde spreading of the aggregates. Furthermore, it allowed development of slices expressing only serine-129 phosphorylation-deficient human α-synuclein (S129G) using an adeno-associated viral (AAV) vector in α-synuclein knockout slices. The processes of aggregation and spreading of α-synuclein were thereby shown to be independent of phosphorylation at serine-129. We provide methods and highlight crucial steps for PFF microinjection and characterization of aggregate formation and spreading. Slices derived from genetically engineered mice or manipulated using viral vectors allow testing of hypotheses on mechanisms involved in the formation of α-synuclein aggregates and their prion-like spreading. |
format | Online Article Text |
id | pubmed-6924077 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-69240772019-12-30 Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates Elfarrash, Sara Jensen, Nanna Møller Ferreira, Nelson Betzer, Cristine Thevathasan, Jervis Vermal Diekmann, Robin Adel, Mohamed Omar, Nisreen Mansour Boraie, Mohamed Z. Gad, Sabry Ries, Jonas Kirik, Deniz Nabavi, Sadegh Jensen, Poul Henning Acta Neuropathol Commun Methodology Article Here we describe the use of an organotypic hippocampal slice model for studying α-synuclein aggregation and inter-neuronal spreading initiated by microinjection of pre-formed α-synuclein fibrils (PFFs). PFF injection at dentate gyrus (DG) templates the formation of endogenous α-synuclein aggregates in axons and cell bodies of this region that spread to CA3 and CA1 regions. Aggregates are insoluble and phosphorylated at serine-129, recapitulating Lewy pathology features found in Parkinson’s disease and other synucleinopathies. The model was found to favor anterograde spreading of the aggregates. Furthermore, it allowed development of slices expressing only serine-129 phosphorylation-deficient human α-synuclein (S129G) using an adeno-associated viral (AAV) vector in α-synuclein knockout slices. The processes of aggregation and spreading of α-synuclein were thereby shown to be independent of phosphorylation at serine-129. We provide methods and highlight crucial steps for PFF microinjection and characterization of aggregate formation and spreading. Slices derived from genetically engineered mice or manipulated using viral vectors allow testing of hypotheses on mechanisms involved in the formation of α-synuclein aggregates and their prion-like spreading. BioMed Central 2019-12-19 /pmc/articles/PMC6924077/ /pubmed/31856920 http://dx.doi.org/10.1186/s40478-019-0865-5 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Elfarrash, Sara Jensen, Nanna Møller Ferreira, Nelson Betzer, Cristine Thevathasan, Jervis Vermal Diekmann, Robin Adel, Mohamed Omar, Nisreen Mansour Boraie, Mohamed Z. Gad, Sabry Ries, Jonas Kirik, Deniz Nabavi, Sadegh Jensen, Poul Henning Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title | Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title_full | Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title_fullStr | Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title_full_unstemmed | Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title_short | Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
title_sort | organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6924077/ https://www.ncbi.nlm.nih.gov/pubmed/31856920 http://dx.doi.org/10.1186/s40478-019-0865-5 |
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