Document Type

Thesis

Date of Completion

Fall 9-27-2021

Department

Biology

Academic Major

Biochemistry and Molecular Biology

Faculty Advisor

Dr. David Donley

Abstract

The proper response to protein signals is necessary for a healthy central nervous system (CNS), and protein dysregulation is a feature of neurodegenerative diseases. Transactive response DNA-binding protein-43 (TDP-43) is an intranuclear protein, but mislocalization is associated with amyotrophic lateral sclerosis (ALS). TDP-43 is released into the extracellular space where it is sensed by microglia, the CNS-resident immune cells. Our data and the literature suggest that microglia respond to TDP-43 dysregulation by increasing CNS inflammation. The goal of this study was to determine the impact of TDP-43 on microglial function and the extent to which microglia recovery. To study the inflammatory response, microglia were stimulated with TDP-43 in a 2x2 factorial design with other inflammatory stimuli. Using iNOS and arginase colorimetric assays, we found that TDP-43 caused microglia to lose the ability to appropriately respond to inflammation. To study recovery, cultured microglia were stimulated with TDP-43 or a vehicle; then the media was changed to remove the stimuli and allow a recovery period. Markers of activation were measured using flow cytometry and metabolic assays. After recovery, microglia had a slight decrease in phagocytic capability as compared to TDP-43 stimulation without recovery. Microglia also demonstrated a metabolic shift toward glycolysis, consistent with a proinflammatory phenotype but returned to baseline levels of metabolic activity after recovery. These data demonstrate that dysregulated TDP-43 shifts the balance of signaling pathways toward an inflammatory phenotype, but microglia begin to recover after removal of the inflammatory stimuli.

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