Profiling Changes in the VTA Dopaminergic Transcriptome during Cocaine Intravenous Self-Administration and Craving

Despite the dramatic rise in cocaine use and overdose deaths, no FDA-approved therapeutics currently exist for treating cocaine use disorder (CUD), in part due to a limited understanding of adaptations in dopamine (DA) neurons, a neuronal cell type crucial for a broad range of behavioral functions. Of note, DA signaling plays a critical role in attributing reward valence and encoding the reinforcing properties of both natural and artificial rewards. To understand the progression of substance use disorders, researchers must investigate the adaptive molecular responses of midbrain DA neurons, particularly to artificial rewards like drugs of abuse such as cocaine. The complex and heterogeneous cellular landscape of the brain, where distinct transcriptional profiles define specific neuronal populations, complicates this investigation. To overcome these challenges, we developed a virus-based method to label and isolate DA nuclei for nuclear RNA-sequencing, specifically targeting ventral tegmental area (VTA) DA neurons. Using a Cre-inducible adeno-associated virus encoding the HA-tagged nuclear envelope protein KASH in DAT-Cre mice, we applied this approach in a mouse model of cocaine intravenous self-administration (IVSA) to explore cocaine-induced transcriptional adaptations in VTA DA neurons.