Authors:

Amelia G Campbell

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About Paper:

Parkinson's disease (PD) is the second most common neurodegenerative disorder affecting approximately 10 million people worldwide. Hallmarks of PD neuropathology include the presence of Lewy body inclusions enriched with aggregated forms of alpha-synuclein (aSyn) and loss of dopaminergic neurons. In the diagnostic field, aSyn seed-amplification assays (SAAs), such as Real-Time Quaking-Induced Conversion (RT-QuIC), are tools that enable sensitive detection of aSyn aggregates in patient derived samples. In RT-QuIC, pathological ?Syn seeds catalyze the aggregation of monomeric ?Syn into fibrils, which bind to Thioflavin T, a dye that fluoresces when bound to amyloid structures. Multiple optimizations of RT-QuIC protocols using human- derived samples are reported in literature. However, there are a lack of studies examining rodent models of PD using SAAs. RT-QuIC is an attractive and underexplored candidate for examining the pathological mechanisms of synucleinopathies through in-vivo models of PD. In effort to improve this model, we aim to use surfactants to reduce spontaneous aggregation of mouse aSyn and accelerate seeded fibril formation. Previous reporting has shown that Triton X (TX-100) keeps aSyn in its monomeric form at the onset of the reaction in human samples. Sodium dodecyl sulfate (SDS) has been described to expose seeds in samples such as cerebral spinal fluid, reducing assay times. In a novel RT-QuIC optimization approach, SDS and TX-100 will be used at varying concentrations to optimize reproducibility and reduce the time of lag phase for rodent-seeded samples. This study aims to establish a rapid and reliable RT-QuIC protocol for preclinical rodent models of synucleinopathies. Keywords: Parkinson's Disease; Synucleinopathies; Alpha-Synuclein; RT-QuIC; Surfactant

Source:

Purdue University / 2025

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Co-authors:

Amelia G Campbell