Project Summary The current project supports the postdoctoral fellow applicant in expanding her technical skills to include in vivo investigations of neurochemical processes during behavioral and pharmacological treatment, as well as ex vivo examination of enzyme activity and receptor coupling. In addition, the applicant will learn analytical chemistry techniques with which to examine these data, including liquid chromatography mass spectrometry (LC-MS), activity based protein profiling (ABPP) and [35S]GTP?S binding. These techniques will be employed in the current proposal, which investigates the effects of stress and protracted alcohol withdrawal on endocannabinoid (eCB) regulation of hypothalamic-pituitary-adrenal (HPA) axis activity and voluntary alcohol consumption. Aim 1 of this proposal targets dysregulation of the eCB system (including catabolizing enzymes and cannabinoid 1 receptor (CB1) binding), GABA release, and HPA responsivity following chronic intermittent ethanol (CIE) vapor exposure. The region of interest in the current studies is the prelimbic (PrL) division of the medial prefrontal cortex since this area exerts top-down control over HPA activity and cognitive behaviors associated with imminent relapse. The second Aim is designed to examine how eCB activity in the PrL affects voluntary alcohol consumption in protracted alcohol withdrawal. The proposed studies will require the applicant to receive extensive methodological training in neurochemical data collection, analysis and interpretation and will support the applicant's overall goal of examining novel therapeutic targets for the treatment of psychiatric illness and alcohol addiction. Furthermore, the applicant's sponsor is highly regarded and successful in the alcohol and eCB fields, and possesses both the fiscal and material resources to support the applicant during the fellowship period. The Scripps Research Institute (TSRI) is an ideal institution for providing additional training and resources to the applicant outside of the laboratory. Departmental meetings as well as meetings of TSRI's Alcohol Research Center (ARC) impart invaluable experience towards participation in research and development goals of the institute, and towards networking in an interdisciplinary and collaborative environment. Additionally, TSRI is committed to promoting the highest standards of the responsible conduct of research, and has partnered with UC San Diego to provide several opportunities towards furthering the applicant's education in these fields. Through these exemplary training opportunities at TSRI, the applicant will be well supported to examine the novel studies proposed in this application.