Date of Award

6-2020

Document Type

Open Access

Degree Name

Bachelor of Science

Department

Biochemistry

First Advisor

Brian Cohen

Keywords

APEX assay, FSHR, effector proteins, infertility, birth control, follicle stimulating hormone, FSH, follicle stimulating hormone receptor, stable transfection, GPCR

Abstract

Many men and women deal with fertility whether it’s needing infertility treatments or contraceptives. The follicle stimulating hormone (FSH) and follicle stimulating hormone receptor (FSHR) play important roles in reproduction and fertility in both males and females. Fertility issues can arise when interactions between FSH and FSHR aren’t functioning properly and FSHR signaling is disrupted. FSHR is a G-protein coupled receptor (GPCR) found on the cell surface of granulosa cells in women and Sertoli cells in men. Activated FSHR initiates a cascade of downstream signaling events that result in different biological effects such as ovarian follicular development and estrogen production in women and sperm production and regulation in men. To create better birth control and fertility treatment options, FSHR signaling is a desirable target. Understanding how the receptor functions can offer insights for how it can be manipulated. Therefore, the effector proteins involved in FSHR signaling must be identified and understood. The APEX Assay has been developed to study these interactions for different GPCRs. This assay uses an engineered ascorbate peroxidase attached to the carboxyl terminus of FSHR which tags proteins within a small radius. Using mass spectrometry, the biotinylated proteins can be quantitatively analyzed to give a better understanding of the proteins associated with FSHR. HEK293 cells are currently being utilized to create a stable cell line expressing the FSHR-APEX receptor. The FSHR-APEX receptor can then be used in the APEX Assay to analyze proteins involved in FSHR signaling, allowing for the creation of birth control methods and fertility treatments that target the FSHR signaling pathways.

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