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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. and Gad, Sabry , et al. (2019) In Acta Neuropathologica Communications 7(1).
Abstract

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... (More)

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.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Alpha-synuclein, Organotypic slices, Prion-like spreading, Serine-129 phosphorylation
in
Acta Neuropathologica Communications
volume
7
issue
1
article number
213
publisher
BioMed Central (BMC)
external identifiers
  • scopus:85077091403
  • pmid:31856920
ISSN
2051-5960
DOI
10.1186/s40478-019-0865-5
language
English
LU publication?
yes
id
8d45566d-cc0c-4047-b27b-8370b655070c
date added to LUP
2020-04-23 10:23:39
date last changed
2024-06-26 14:31:12
@article{8d45566d-cc0c-4047-b27b-8370b655070c,
  abstract     = {{<p>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.</p>}},
  author       = {{Elfarrash, Sara and Jensen, Nanna Møller and Ferreira, Nelson and Betzer, Cristine and Thevathasan, Jervis Vermal and Diekmann, Robin and Adel, Mohamed and Omar, Nisreen Mansour and Boraie, Mohamed Z. and Gad, Sabry and Ries, Jonas and Kirik, Deniz and Nabavi, Sadegh and Jensen, Poul Henning}},
  issn         = {{2051-5960}},
  keywords     = {{Alpha-synuclein; Organotypic slices; Prion-like spreading; Serine-129 phosphorylation}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{BioMed Central (BMC)}},
  series       = {{Acta Neuropathologica Communications}},
  title        = {{Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates}},
  url          = {{http://dx.doi.org/10.1186/s40478-019-0865-5}},
  doi          = {{10.1186/s40478-019-0865-5}},
  volume       = {{7}},
  year         = {{2019}},
}