RESEARCH SUMMARY Alcohol use disorder (AUD) is characterized by the inability to control or stop the use of alcohol, even in the face of negative consequences or after long-term abstinence. Thus, it is important to understand the risk factors that contribute to this loss of control such as environmental cues that can contribute to an increase in probability of relapse, compulsive drinking, and withdrawal symptoms after abstinence. Previous research has shown that ventral tegmental area (VTA) dopamine (DA) neurons can modify the learned value of reward- associated cues to alter reward-seeking behavior by driving an increase cue reactivity and cue-driven reward- seeking. It is also known that VTA DA neuron activity increases compulsive alcohol seeking and cue-induced relapse of alcohol seeking. However, which receptors control the activity of VTA DA neurons on cue-related behaviors have yet to be determined. Systemic GABAB receptor activation is shown to reduce the mentioned alcohol-related behaviors in rodents in addition to reducing the ability of cues to reinstate alcohol seeking. Baclofen, a direct GABAB receptor agonist, has been shown to have a dose-related reduction in these behaviors while related studies showed that the VTA was implicated as baclofen’s key site of action due to intra-VTA injections having suppression of cue-elicited reward seeking in rats. Yet, the specific neuronal populations in the VTA responsible for baclofen’s effects remain unknown. Thus, the broad goal is of the proposal is test the significance of a potential molecular target from a reward-related neuron population on alcohol cue reactivity, cue-evoked aversion-resistant drinking and relapse plus on baclofen’s seen effects. The objective of this proposal is to investigate the impact of deleting GABAB receptors in VTA DA neurons on cue-evoked relapse (Aim 1) and aversion-resistant drinking (Aim 2). Additionally, to test the role of these receptors on the previously seen effects of baclofen with the addition of intra-VTA microinjections of baclofen. Given that these receptors and neuron population have been shown to influence these behaviors in rodents, I hypothesize that GABAB receptors in VTA dopaminergic neurons are responsible for constraining the cue-elicited alcohol-related behaviors and for the previously seen baclofen induced effects. To test this, I will use the cutting-edge CRISPR-Cas9 approach by bilaterally microinjecting high-titer AAV viral vectors harboring guide RNAs targeting GABAB receptors or control (Rosa), and a Cre-dependent GFP for visualizing expression (Experiment 1A and 2A). Additionally, a separate subject group will also have a bilateral guide cannula implant for intra-VTA baclofen delivery (Experiment 1B and 2B). Following this, the aforementioned behaviors will be tested and an analysis of lever presses and g per kg of alcohol consumed will take place. That said, during this proposal I will gain valuable technical and theoretical training in genetic manipulation, behavioral methods, histological processing, imaging techniques, intracranial surgeries, and computational analysis. Therefore, allowing me to become a successful independent addiction neuroscientist.

PROJECT NARRATIVE Alcohol use disorder is characterized by the inability of controlling or stopping alcohol use, even in the face of negative consequences or after long-term abstinence. Research shows that ventral tegmental area (VTA) dopaminergic neurons drive reward-seeking behavior and increased value of reward-associated cues while related studies show GABAB receptors and baclofen constraining behaviors like cue-induced reinstatement and compulsive alcohol use in the presence of an aversive stimulant. However, which receptors control the activity of VTA dopamine neurons on cue-related behaviors and which specific neuronal populations in the VTA are responsible for baclofen’s effects has yet to be determined, therefore, we are proposing test the role of GABAB receptors in VTA dopamine neurons on cue-induced reinstatement, aversion resistant drinking, and on the previously seen effects of baclofen.