Protocol for using an ELISA to detect total α-synuclein levels in Drosophila melanogaster lines expressing human α-synuclein point mutations.
Protocol for using an ELISA to detect total α-synuclein levels in Drosophila melanogaster lines expressing human α-synuclein point mutations.
Where did the research take place?
The study site has not been established. Author addresses may differ from where the research occurred.
San Antonio, US · Author affiliation
Department of Cellular and Integrative Physiology, University of Texas San Antonio Health Science Center, Joe R. and Teresa Lozano Long School of Medicine, San Antonio, Texas, USA.Location evidence
Perry, US · Author affiliation
Perry and Ruby Stevens Parkinson's Disease Center of Excellence, University of Texas San Antonio Health Science Center, Joe R. and Teresa Lozano Long School of Medicine, San Antonio, Texas, USA.Location evidence
Publication status: preprint
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Original abstract
In Parkinson's disease (PD), aggregation of alpha-synuclein (α-syn) contributes to neuronal dysfunction and death, particularly in dopaminergic neurons, driving disease progression. Several pathogenic point mutations have been identified in human α-syn such as A30P, E46K, H50Q, G51D, A53T and A53E that have been associated with PD. In this study, a sandwich ELISA assay was developed to quantify total α-syn levels in various Drosophila melanogaster genotypes expressing A30P, E46K, G51D and A53T. Using this approach, we compared α-syn levels across wild-type (WT) and mutant forms of the protein. The E46K and A53T mutations exhibited higher total α-syn concentrations compared to WT and the G51D mutation. In addition to characterizing mutationdependent differences in α-syn levels, this assay was applied to evaluate smallmolecule modulators that inhibit α-syn aggregation. These findings demonstrate that the ELISA-based approach provides a useful platform for quantifying α-syn and assessing potential therapeutic compounds targeting αsyn pathology.