Extended supplementary data for manuscript: Gene regulatory mechanisms of cellular memory of a single cocaine exposure in VTA dopaminergic neurons

Drug addiction is a chronic brain disease with major global health and economic costs. Many drugs of abuse hijack reward circuitry, reinforcing drug-taking behaviour. While drug effects on synaptic plasticity are transient, the gene regulatory mechanisms driving long-term memory of drug exposure remain unclear. We assessed gene expression and chromatin accessibility in ventral tegmental area dopamine neurons (VTA-DNs) in a time-course following acute cocaine exposure. Expression changes persist for 14 days, including upregulation of addiction-associated neuropeptides, while AP-1 chromatin binding sites shift from early stimulus-driven activity to sustained silencing. We show downregulation of Foxa2 and its targets, followed by erosion of DN identity lasting up to 14 days, and sustained upregulation of Polycomb-repressed genes. We show that a single cocaine exposure is sufficient to trigger enduring transcriptional and epigenetic changes in reward-associated neurons as memory of drug exposure, and provide a high-resolution temporal map of cocaine-induced regulatory plasticity as groundwork for finding molecular targets that drive addiction onset.