Expressing Alpha-Synuclein in RCSN-3 Cells to Improve the Study of Pathways in Parkinson’s Disease

Alpha-synuclein (αS) is a dopamine-regulating protein in the brain that can form aggregates, clusters of misfolded proteins, 和 cause neurodegeneration in the form of Parkinson’s Disease (PD). This ongoing research project explores the furthered development of the RCSN-3 cell line where it can express fluorescently tagged human αS. The development of this in vitro model will necessitate transfection of αS DNA, 荧光显微镜, 和 differentiation of the cells into neurons, with the ultimate goal of having a better model to both screen small-molecule drugs that could inhibit αS aggregation 和 study other related pathways, such as the polyamine pathway. With an efficacious working model of dopaminergic neurons, we can now work to replicate the results of cell viability 和 aggregation assays accomplished by our lab in past years to show the applicability of the findings to the pathogenesis of PD.
We went about furthering the RCSN-3 cell line using DNA plasmid transfection protocols, seeding of preformed αS fibrils into confluent cell culture, 和 荧光显微镜 to establish the viability of the cell line as a model. Our next steps involve differentiating the cells into neurons 和 replicating assays with previously established results to exp和 on data implicating the polyamine pathway in PD pathogenesis, as well as obtaining samples to send for Proteomics 和 Metabolomics analysis.