Single nuclei RNAseq of rat area nucleus accumbens after morphine exposure

We performed single-nuclei RNAseq of Brown Norway rat nucleus accumbens after a single injection of morphine or after morhpine self-administration Overall design: We performed the 10x Genomics 3' gene expression assay (v3.1) on nuclei isolated from frozen Brown Norway (BN) rat nucleus accumbens brain punches. A cohort of male BN rats received an experimenter delivered dose of morphine (10.0 mg/kg, ip) or saline (n=5 each) and were sacrificed two hours after injection. A second cohort of male BN rats underwent volitional IV morphine self-administration (1.0mg/kg/infusion) for three hours daily for 10 days, along with their saline yoked controls (n=6 each), and animals were sacrificed two hours after their final self-administration session. Brains were removed and snap-frozen in dry ice-cold isopentane. Brains were coronally sectioned and the nucleus accumbens core and shell was removed by micro-punch bilaterally, and the punches were stored at -80°C until further processing. Nuclei were isolated by differential centrifugation and microfluidically captured using the 10x Genomics Chromium controller. Sequencing libraries were constructed and sequenced on an Illumina NovaSeq 6000.