snRNA-seq dataset for "Morphine regulates astrocyte transcriptional dynamics by stimulation of glucocorticoid signaling"
This dataset contains single-nucleus RNA-seq results from rat ventral tegmental area brain tissue in a chronic pain model with morphine administration. Overall design: This experiment contains 28 biological samples, which represent four experimental groups of rats. To assess neuropathic pain response, Complete Freund's Adjuvant (CFA) or saline vehicle (100 µl) was injected into the plantar surface of one hind paw of male and female rats (n = 3-4/sex/treatment group). Mechanical sensitivity was then evaluated 48 hours and 7 days after the injection using the Von Frey assay. 7 days after CFA injection, rats received either morphine (10mg/kg) or saline injections I.P., generating a 2x2 experimental design (Veh-Sal, Veh-Mor, CFA-Sal, CFA-Mor). One hour after injections, animals were euthanized prior to rapid decapitation. Brains were blocked in 1mm sections, and VTA tissue was obtained and flash frozen, and stored at -80°C prior to nuclei isolation. Following FACS-assisted nuclei isolation and quantification, a total of 8,000 nuclei pooled from two rats (4,000 male nuclei/4,000 female nuclei from the same treatment group) and were loaded into individual wells of the Chromium NextGem Single Cell Chip (10x Genomics, catalog no. 10000121) and processed on the 10X Genomics Chromium Controller. Libraries were constructed according to manufacturer's instructions for Chromium Next GEM single cell 3' library and gel bead kit (10x Genomics, v3.1 single index, catalog no. 10000121, version 3 chemistry). Libraries were sequenced on an Illumina NovaSeq6000 (S2 flow cell)  to an average read depth of ~42,000 read pairs per nuclei. Forward and reverse paired-end reads are denoted as "R1" or "R2" in raw file names.