Project Summary Pharmacology is the study of how medicine works to produce a desired therapeutic outcome. As medicines are developed, they are designed to target specific receptors within the body and alter the way cells function. Of these drug targets, G-protein coupled receptors (GPCRs) are the most common, and my research examines how GPCRs function once drugs bind to them. GPCRs alter cellular responses by initiating G-protein signaling. This signaling was thought to occur only at the surface of cells, but emerging research has shown that G-protein signaling occurs within the cell as well. Therefore, it is important to understand how GPCR trafficking to and from subcellular locations impact their overall signaling capabilities. The cannabinoid receptor-1 (CB1) is a GPCR heavily expressed throughout the brain. It plays numerous neurobiological roles and has been a potential drug target in managing appetite, depression, pain and anxiety. However, drugs that bind to CB1, also known as cannabinoids, produce a wide variety of unintended side effects. For example, THC, the psychoactive component of cannabis, is a cannabinoid that is heavily associated with drug use and substance abuse disorders. My research is particularly focused on understanding how cannabinoids that bind to CB1 produce different biological outcomes. I aim to expand their value as promising therapeutic targets by studying how cannabinoids regulate CB1's subcellular activity. Understanding the functional roles of intracellular GPCR signaling is a new and exciting area of pharmacology and therapeutic research where drugs can be specifically designed for intracellular targeting of receptors. This could have significant implications for understanding the varying outcomes of CB1 activation, as CB1 exhibits significant subcellular localization when compared to GPCRs that are most commonly studied. However, the significance of CB1's subcellular localization requires further investigation. The proposed research project will investigate how CB1 is differentially activated by cannabinoids within the brain, while simultaneously delineating the spatial components of CB1 activation. The results gained from completion of this project will provide a framework for developing CB1 as a viable therapeutic target.