Cell-type specific central amygdala neurotransmission in alcohol dependence

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Project Summary/Abstract: Research: Alcohol use disorder is highly prevalent and costly, and only a few modestly effective pharmacological treatments are available. Current evidence indicates that repeated consumption of alcohol induces changes in gene expression within neurons in the central amygdala (CeA), and as animals become alcohol dependent, these changes drive pathological alcohol consumption. For example, pharmacological studies have shown that repeated alcohol exposure increases corticotropin-releasing factor (CRF) signaling in the CeA, which promotes excessive alcohol consumption in rodents. However, the source of this CRF is not certain and it is also unlikely that increased CRF signaling is the sole driver of increased alcohol consumption in these animals. This project will test the hypothesis that neurons within the lateral CeA are the source of this CRF and that these neurons produce additional neuropeptides that drive excessive alcohol drinking. This hypothesis will be tested by examining drinking behavior after individually downregulating the neurotransmitters CRF, dynorphin, or GABA in CeA-CRF neurons using a BAC transgenic Crh-Cre rat and Cre-dependent RNA interference. The project will also use an unbiased approach to explore alcohol-induced gene expression in all neurons of the CeA by using single nuclei RNA-seq to identify neuronal populations and the genes within these populations that are most affected as animals become alcohol dependent. The results of this work will lead to new, testable hypotheses about specific proteins that could be targeted to reduce excessive drinking in alcohol dependent individuals. Training: This research will train the applicant in several experimental techniques including rodent drinking behavior, testing and validation of genetic tools, single-nuclei sequencing, RNA quantification, and use of bioinformatics analysis tools. Through this training, the applicant will develop expertise in the use of molecular tools to manipulate gene expression and in bioinformatics approaches to analyze transcriptomes. The training program will also develop professional skills, including scientific writing and presentation, and programming, to further the applicant’s goal of becoming an independent academic neuroscientist.