Modeling ?-Synucleinopathy in Organotypic Brain Slice Culture with Preformed ?-Synuclein Amyloid Fibrils.

ABSTRACT: BACKGROUND:Synucleinopathy is a group of neurodegenerative disorders characterized by neurodegeneration and accumulation of alpha-synuclein (?-syn) aggregates in various brain regions. The detailed mechanism of ?-syn-caused neurotoxicity remains obscure, which is partly due to the lack of a suitable model that retains the in vivo three-dimensional cellular network and allows a convenient dissection of the neurotoxic pathways. Recent studies revealed that the pre-formed recombinant ?-syn amyloid fibrils (PFFs) induce a robust accumulation of pathogenic ?-syn species in cultured cells and animals. OBJECTIVE:Our goal is to determine whether PFFs are able to induce the pathogenic ?-syn accumulation and neurotoxicity in organotypic brain slice culture, an ex vivo system that retains the in vivo three-dimensional cell-cell connections. METHODS/RESULTS:Adding PFFs to cultured wild-type rat or mouse brain slices induced a time-dependent accumulation of pathogenic ?-syn species, which was indicated by ?-syn phosphorylated at serine 129 (p?-syn). The PFF-induced p?-syn was abolished in brain slices prepared from ?-syn null mice, suggesting that the p?-syn is from the phosphorylation of endogenous ?-syn. Human PFFs also induced p?-syn in brain slices prepared from mice expressing human ?-syn on a mouse ?-syn-null background. Furthermore, the synaptophysin immunoreactivity was inversely associated with p?-syn accumulation and an increase of neuronal loss was detected. CONCLUSION:PFF-treatment of brain slices is able to induce key pathological features of synucleinopathy: p?-syn accumulation and neurotoxicity. This model will be useful for investigating the neurotoxic mechanism and evaluating efficacy of therapeutic approaches.

SUBMITTER: Roux A 

PROVIDER: S-EPMC7683096 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature