“ … alkalinity range based on color change of the indicator. Alexandra Temple ‘20 … ”
Abstract
Samples of water were collected from three different sites: Delanson Pond, Tap Water, and Saratoga Springs spring water. These samples were analyzed in an effort to determine the concentrations of inorganic ions and metals present in the samples. Goals: Analyze the water samples and compare their respective concentrations of barium, lead, calcium, chloride, and sodium.
Follicle stimulating hormone (FSH) and its receptor (FSHR) play important roles in reproduction and fertility in both men and women. Fertility problems arise when FSH, FSHR, or any downstream signaling components don't function properly. FSHR is a G-protein coupled receptor (GPCR) found on the cell surface of granulosa cells in women and Sertoli cells in men. When activated by FSH, FSHR initiates a signaling cascade that can result in downstream results such as ovarian follicular development and estrogen production in women and sperm production in men. To better understand how the receptor functions, the interactions between FSHR and effector proteins involved in signaling must be identified and understood. We plan to study the interactions between FSHR and its effector proteins using the APEX Assay. This assay utilizes an engineered ascorbate peroxidase (APEX) which is attached to the C-terminus of FSHR. APEX modifies proteins using biotin phenol and hydrogen peroxide, which creates a biotin-radical, thus labeling proteins within a small radius of FSHR because of the short lived nature of the radical. Using mass spectrometry, the biotinylated proteins can be quantitatively analyzed to give a better understanding of the proteins associated with FSHR and the changes in the proteins over time after hormone treatment. Using HEK293 cells, we created a stable cell line expressing an FSHR-APEX construct. We are currently using the FSHR-APEX expressing cells in the APEX Assay to analyze the proteins that are involved in FSHR signaling. This research will give us a better insight as to what effector proteins interact with FSHR and how these interactions change following hormone stimulation. This will provide us with a better understanding as to how the receptor signals and initiates downstream signaling and how this might affect fertility in humans.
“ … Abstract Temple,Alexandra Investigating Human Follicle Stimulating Hormone … Hormone Receptor and its Partners Using the APEX Assay By Alexandra Temple ********** Submitted in partial fulfillment … ”
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.
