Biochemistry and Molecular Biology-BS
Biochemistry and Molecular Biology
Dr. David Sheridan, Ph.D.
First Committee Member
Dr. Jennifer Bennett, Ph.D.
Second Committee Member
Dr. David Robertson, Ph.D.
Calcium Channels, Glucocorticoid Receptor, Cortisol, Stress
Biochemistry, Biophysics, and Structural Biology | Molecular Biology
L-type calcium channels couple membrane depolarization to muscle contraction and aid in signal conduction between cells. Cav1.3 calcium channel isoform is expressed in neurons and endocrine cells. Nervous and endocrine tissues also express both Glucocorticoid (GR) and Mineralocorticoid receptors (MR) that are responsible for binding serum cortisol to induce immediate and long term changes within the cell. While neurons co-express both GRs and MRs, GR activation occurs in response to high concentrations of cortisol. There is a direct correlation between GR activation and the amplitude of calcium currents (Champeau, 2007). To determine potential interactions between the GR and the calcium channel isoforms, we will co-express a GFP-tagged GR, purchased from Addgene, and a mRubyC1-tagged L-type calcium channel in a simplified, non-native cellular system, Chinese Hamster Ovary (CHO) cells. Specifically targeted primers were designed to clone the Cav1.3 sequence in preparation for its insertion into the mRubyC1 red fluorescent vector. The Cav1.3+mRubyC1 construct was then transfected into CHO cells expressing GFP-tagged GR. Injection of a high concentration of cortisol into the cellular environment will activate the GR receptor, and fluorescence capture imaging will determine if there is an in vitro interaction between Cav1.3 and GR.
Soska, Mallory, "Potential Interactions Between Glucocorticoid Receptors and L-Type Cav1.3 Ion Channels Expressed in Chinese Hamster Ovary (CHO) Cells" (2019). Distinction Papers. 70.
Available for download on Sunday, April 16, 2023