Date of Award

Fall 12-2017

Degree Type

Thesis

Degree Name

MS Microbiology

Department

Biology

Advisor

Heping Zhou, Ph.D.

Committee Member

Daniel B. Nichols, Ph.D.

Committee Member

Constantine Bitsaktsis, Ph.D.

Committee Member

Angela Klaus, Ph.D.

Keywords

palmitate, Nuero2A, GLUT, GPR

Abstract

Palmitate (PA) is a saturated fatty acid that has been reported to be elevated in individuals with metabolic syndromes. The effects of fatty acids on the expression of various fatty acid receptors on neuronal cells is unknown, as is the effects of fatty acids on GLUT transporters. In this study, Neuro-2a (N2a) cells were treated with either 300μM of bovine serum albumin (BSA) or 300μM of palmitate conjugated to BSA (PA) at various time points to determine the effect of palmitate on the expression of various fatty acid receptors and GLUT transporters. RNA isolation followed by RT-PCR with primers specific to various G- protein coupled receptors (GPRs) and GLUT transporters was performed on these treated cells and further analyzed with statistical tests. 24 hour treatment with 300μM PA yielded increased mRNA expression of GPR41, GPR84, GPR119, GPR120 and GLUT5 when compared to vehicle control. In order to determine how GLUT5 may be regulated, N2a cells were treated with 300μM BSA along with PKA inhibitor KT5720 or GPR120 agonist GW9508. RNA isolation followed by RT-PCR showed that GPR41, GPR119 and GLUT5 expression increased with PKA inhibition, and GPR120 expression increased with GPR120 stimulation. These results suggest that PKA may play a role in the regulation of GPR41, GPR119 and GLUT5 in neuronal cells stimulated with palmitate.

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