Department or Program
Primary Wellesley Thesis Advisor
Glutamate transporters (GLTs) remove glutamate from synapses to prevent excitotoxicity and maintain conditions necessary for glutamatergic signaling. Once transported into cells adjacent to the synapse, glutamate can be metabolized into α-ketoglutarate (α-KG) or glutamine. Since GLT deletions are often lethal in mammals, C. elegans were used to investigate the role of GLTs on metabolism and aging in this thesis. The lifespans of C. elegans GLT-knockouts (GLT-1;GLT-3, GLT-4, and GLT-5) were measured and compared to wildtype worms. A well-established method involving FUdR, a chemical that prevents egg-laying in C. elegans, was used to conduct the lifespan experiments. Results indicated that GLT-4 mutants had shorter lifespans, GLT-5 mutants had longer lifespans, and GLT-1;GLT-3 mutants had similar lifespan lengths compared to wildtype worms. Despite being the primary method for aging experiments in C. elegans, FUdR is costly and hazardous. We pursued an alternative lifespan technique using mesh-well plates to separate C. elegans from their laid eggs. While the mesh-well technique had multiple theoretical advantages, this technique did not consistently separate adult worms from laid-eggs. To determine the influence of glutamate metabolism on the lifespan differences between strains, we refined a protocol to analyze the levels of glutamine, glutamate, and α-KG metabolites using 1H-NMR. While NMR could not be used to measure metabolite levels in C. elegans samples, metabolite experiments are on-going. These findings bring us closer to understanding how the effects of glutamate metabolism in cellular respiration and neurotransmission influence aging.