Single nucleus RNA-seq of rat nucleus accumbens

Single nucleus RNA-seq of nucleus accumbens from male Brown Norway rats after a single acute or repeated saline injections 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 saline (n=5) and were sacrificed two hours after injection. A second cohort of male BN rats received repeated saline injections as yoked controls in a study of morphine for three hours daily for 10 days (n=6), and animals were sacrificed two hours after the final session. Brains were removed and snap-frozen in dry ice-cold isopentane. Brains were coronally sectioned and the nucleus accumbens core and shell were removed by micro-punch bilaterally, and the punches were stored at-80°Cuntil 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.