ENGINEERING GENETICALLY ENCODED FLUORESCENT BIOSENSORS TO STUDY THE ROLE OF MITOCHONDRIAL DYSFUNCTION AND INFLAMMATION IN PARKINSON’S DISEASE

Abstract

Parkinson’s disease is a neurodegenerative disorder\ncharacterized by a loss of dopaminergic neurons, where mitochondrial\ndysfunction and neuroinflammation are implicated in this process. However, the\nexact mechanisms of mitochondrial dysfunction, oxidative stress and\nneuroinflammation leading to the onset and development of Parkinson’s disease\nare not well understood. There is a lack of tools necessary to dissect these\nmechanisms, therefore we engineered genetically encoded fluorescent biosensors\nto monitor redox status and an inflammatory signal peptide with high\nspatiotemporal resolution. To measure intracellular redox dynamics, we\ndeveloped red-shifted redox sensors and demonstrated their application in dual\ncompartment imaging to study cross compartmental redox dynamics in live cells.\nTo monitor extracellular inflammatory events, we developed a family of\nspectrally diverse genetically encoded fluorescent biosensors for the\ninflammatory mediator peptide, bradykinin. At the organismal level, we characterized the locomotor effects of mitochondrial toxicant-induced\ndopaminergic disruption in a zebrafish animal model and evaluated a behavioral\nassay as a method to screen for dopaminergic dysfunction. Pairing our\nintracellular redox sensors and our extracellular bradykinin sensors in a\nParkinson’s disease animal model, such as a zebrafish toxicant-induced model will\nprove useful for dissecting the role of mitochondrial dysfunction and\ninflammation in Parkinson’s disease.