Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates

TY - JOUR

T1 - Organotypic slice culture model demonstrates inter-neuronal spreading of alpha-synuclein aggregates

AU - Elfarrash, Sara

AU - Jensen, Nanna Møller

AU - Ferreira, Nelson

AU - Betzer, Cristine

AU - Thevathasan, Jervis Vermal

AU - Diekmann, Robin

AU - Adel, Mohamed

AU - Omar, Nisreen Mansour

AU - Boraie, Mohamed Z

AU - Gad, Sabry

AU - Ries, Jonas

AU - Kirik, Deniz

AU - Nabavi, Sadegh

AU - Jensen, Poul Henning

PY - 2019/12/19

Y1 - 2019/12/19

N2 - 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.

AB - 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.

KW - Alpha-synuclein

KW - Organotypic slices

KW - Prion-like spreading

KW - Serine-129 phosphorylation

U2 - 10.1186/s40478-019-0865-5

DO - 10.1186/s40478-019-0865-5

M3 - Journal article

C2 - 31856920

SN - 2051-5960

VL - 7

JO - Acta Neuropathologica Communications

JF - Acta Neuropathologica Communications

IS - 1

M1 - 213

ER -