The Unconventional Secretion of Alpha-Synuclein by Autophagic Mechanisms and the Novel Characterization of Pathological Alpha-Synuclein Associated Extracellular Vesicles

Synucleinopathies are a group of neurodegenerative diseases associated with the intracellular accumulation of aggregated α-synuclein, a neurotoxic process resulting in cell death. Research indicates that pathology results from α-synuclein misfolding and self-propagation, as well as, cell-to-cell transmission. Furthermore, pathological α-synuclein transmission may occur through unconventional secretion. In this thesis, we investigated how autophagy(definition) influences unconventional secretion of our novel α-synuclein dual-split protein model that we first verified by reproducing documented results from treatment of known autophagic regulators. Afterwards, we investigated how knocking out autophagic proteins ATG7 and Beclin-1 influenced α-synuclein secretion. Additionally, we developed a methodology for analyzing extracellular vesicles on a single vesicle level using microscopy and immunofluorescent staining which we demonstrate by analyzing the vesicles secreted from SH-SY5Y cells treated with sonicated preformed α-synuclein fibrils.