Alcohol-Induced Alternative Splicing in Drosophila Memory Circuits

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Repeated alcohol experiences can produce long-lasting memories for sensory cues associated with intoxication. These memories can problematically trigger relapse in individuals recovering from alcohol use disorder (AUD). The molecular mechanisms by which ethanol changes memories to become long-lasting and inflexible remain unclear. We recently demonstrated that formation of these memories results in expression of alternative transcript isoforms in memory-encoding neurons in Drosophila melanogaster. Drosophila rely on mushroom body (MB) neurons to make associative memories, including memories of ethanol-associated sensory cues. Decreasing expression of genes that play a role in splicing in adult MB neurons reduces formation of these memories, demonstrating the necessity of RNA processing in ethanol memory formation. Moreover, decreasing expression of genes that are alternatively spliced like the Dopamine-2-like Receptor (Dop2R) in adult MB neurons reduces ethanol memory formation. This suggests that the splicing changes in these genes has functional implications for future memory formation. The central hypothesis of this proposal is that the dynamic alternative splicing of Dop2R in response to alcohol exposure directs reward behavior by changing the activity of neural circuits. We generated mutant Drosophila that have forced expression of the naïve or trained Dop2R isoform, which allows us to interrogate how these splice variants affect neural dynamics and behavior at a highly mechanistic level. The applicant will reveal intracellular localization of DOP2R protein (Aim 1, how alternative splicing of Dop2R affects reward memory (Aim 2), and plasticity within memory circuits (Aim 3). By completing these aims, the applicant will gain scientific and technical expertise in cellular neurobiology and neurogenetics, behavioral assessment of Drosophila, super-resolution imaging, and 2-photon calcium imaging. Through a comprehensive training plan, this fellowship will facilitate the professional development of the applicant in robust experimental design and data analysis; written and oral scientific communication; and effective and inclusive mentoring. Successful completion of the project and training goals are fully supported by the interactive and supportive institutional environment of Brown University and in the Neuroscience Graduate Program, and will prepare the applicant for the next steps towards an independent scientific career.