Undergraduate Honors Thesis Projects

Date of Award

Spring 4-14-2017

Document Type

Honors Paper

Degree Name

Biochemistry and Molecular Biology-BS

Department

Biochemistry and Molecular Biology

Advisor

Dr. David Sheridan

First Committee Member

Dr. John Tansey

Second Committee Member

Dr. Joby DeCoster

Keywords

Stress, Glucocorticoid Receptor, HEK cell, Cortisol

Subject Categories

Molecular and Cellular Neuroscience

Abstract

In this study, we aimed to investigate the interaction between glucocorticoid receptors and calcium channels. To begin our research, we began to optimize the transfection efficiency of a glucocorticoid receptor within human embryonic kidney cells. Results found that this non-native cell was able to be transfected with pK7-GR-GFP DNA. Although it was unexpected, we were unable to obtain a transfection efficiency of 10% to move forward with other planned experiments. However, through our efforts, we were able to achieve a transfection efficiency of 0.04% and troubleshoot many difficulties that prevented obtaining a higher transfection efficiency. It was found that the QIAgen kit we used to purify the plasmid DNA of the glucocorticoid receptor retained some of the DNA within the membrane of the QIAprecipitator Module when eluting the DNA. Although we were not able to reach optimal conditions, we have outlined experiments that we hope to complete in the future. These experiments require subjecting the transfected cells to cortisol and the cotransfection of the human embryonic kidney cells with the DNA of a glucocorticoid receptor and calcium channel. Through these experiments, we would be able to better understand how glucocorticoid receptors are affected by cortisol and their association with other proteins, such as calcium channels. This would allow us to gain a better understanding of how these proteins might affect the excitability of neurons.

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