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

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thesis
posted on 10.06.2019, 16:39 by Stevie Norcross

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

Funding

NSF graduate research fellowship (DGE-1333468)

NIH R21 EY026425

History

Degree Type

Doctor of Philosophy

Department

Chemistry

Campus location

West Lafayette

Advisor/Supervisor/Committee Chair

Dr. Mathew Tantama

Additional Committee Member 2

Dr. Mary Wirth

Additional Committee Member 3

Dr. Angeline Lyon

Additional Committee Member 4

Dr. Chittaranjan Das