The accumulation of misfolded alpha-synuclein (α-syn) into insoluble aggregates is a molecular characteristic in several neurodegenerative diseases, in particular, Parkinson’s disease, multiple system atrophy and dementia with Lewy bodies. Recent studies have shown that once α-syn has started to misfold and aggregate the tendency is transmitted from cell-to-cell, thereby recruiting endogenous α-syn and inducing fibrillization in a prion-like manner. Similar to prion diseases, the heterogeneity of protein aggregate conformations may explain the variable efficiency of template fibrillization and propagation of these misfolded proteins. Additionally, the type of structural strain might be disease defining.
In this project, we focus on the cell-to-cell spreading of defined preformed α-syn seeds. Thereby observing the role of the structural features in spreading efficiency, localization in cells and recruitment of endogenous α-syn.
We want to determine the importance of different types of α-syn seeds in the spreading and transmission of α-syn misfolding between cells. We utilize a microfluidic system with spatially separated subcultures linked by channels through which neurites of a model system can grow. This allows us to study how and when an α-syn seed-exposed “donor” cell takes up the seed and transmits it through neurite contacts to the separated “recipient” cell. For the structural analysis of the seeds and induced fibrils, our model system will be integrated in the in-house developed visual proteomics pipeline to allow single-cell analysis and high-resolution cryo-EM.