Date of Award
Master of Science
Dr. Charles Ide
Dr. John Spitsbergen
Dr. Robert Eversole
Multiple System Atrophy, PCR synucleinopathy, Alpha Synuclein, Parkinson's Disease
Masters Thesis-Open Access
Multiple System Atrophy (MSA) is a rare neurological disease that mainly implicates the neuronal protein, alpha synuclein, as being centrally involved in MSA pathology. Despite synucleinopathy dogma, little is known about alpha synucleins role in MSA pathogenesis. The Ide laboratory previously conducted an Affymetrix gene expression study utilizing the post-mortem pons tissue from eight individuals with MSA and five individuals without any known neurological disorders. One conclusion from that study was that downregulation of numerous mitochondrial homeostasis related genes had occurred. It is hypothesized that alpha synuclein can perturb mitochondrial homeostasis through direct interaction with mitochondria. This study utilized Reverse Transcriptase PCR and Droplet Digital PCR to confirm downregulation of the mitophagy related gene, Bnip3, that was observed in the MSA Affymetrix study, when C6 rat glioma cells were treated with a single dose of human wild type alpha synuclein. qPCR and ddPCR was then used to assess the impact of alpha synuclein treatment on mitochondrial genome copy numbers, which served as a surrogate measure of mitochondrial number, and the single copy nuclear-encoded gene, Beta Actin. Detection of mtDNA and Beta Actin was decreased at all time points assessed. This study demonstrates that a single treatment of alpha synuclein resulted in decreased detection of mitochondrial genomes per cell, which offers potential insight into synucleinopathy pathogenesis.
Helmus, Michael John, "PCR Analysis on the Effects of Alpha Synuclein Treatment on C6 Rat Glioma Cells" (2019). Master's Theses. 5103.