Infertility affects millions of individuals worldwide and the causes of infertility are often unexplained. There are a number of signaling pathways that are involved in reproduction and defects in these pathways could contribute to infertility. Specifically, we are interested in studying how FSHR (follicle stimulating hormone receptor) responds to human follicle stimulating hormone (hFSH) because having the ability to enhance or downregulate its signaling has implications for fertility treatments. When FSH binds to its cognate G-protein-coupled receptor (GPCR) it activates the production of cAMP as a second messenger. Previous work in our lab has demonstrated that FSHR needs to be embedded in a specialized microdomain of the cellular membrane called a lipid raft to mediate this response. After treating FSHR-expressing cells with lipid raft-disrupting drugs, decreased cAMP was observed as a decrease in activated (phosphorylated) cAMP response-element binding protein (pCREB) within the Gα/cAMP/PKA pathway. The amount of pCREB is an indirect measurement of cAMP production. To establish a direct relationship between FSHR signaling and cAMP production, a quantitative luciferase assay was developed. Luciferase is an enzyme found in fireflies that allows them and other bioluminescent organisms to glow. A cell line expressing hFSHR (HEK293-hFSHR) was transiently transfected with a plasmid to express luciferase under the control of multiple, tandem cAMP response elements. This acts as a quantitative reporter for cAMP because it produces more luminescence as the amount of cAMP in the cells increases. Cells were treated with Methyl-β-cyclodextrin (MβCD) which acts as a lipid raft-disrupting agent by depleting cholesterol from cells and destroying lipid rafts. All cells were stimulated with various amounts of FSH to create a dose-response curve. MβCD-treated cells produced less cAMP in a dose-dependent manner compared to the untreated cells. These results differ from previous observations of the FSH-induced activation of p44/42 MAP kinase (ERK1/2). It was found that treatment with the lipid raft disrupting agents resulted in increased basal cAMP production (as measured by pCREB activation). Future studies will test different lipid raft-disrupting agents to further investigate the role of lipid rafts in FSH induced cAMP production from FSHR stimulation. Studying and characterizing FSHR behavior is essential to furthering our understanding of infertility and improving the possible treatments for both men and women who are struggling to conceive.