Mimicking opioid analgesia in cortical pain circuits

We performed single nuclei transcriptomics on the anterior cingulate cortex of mice cohorts that received either neuropathic pain (sciatic nerve injury) or sham surgery, followed by varying posto-injury periods (3 days, 3 weeks, or 3 months). Five minutes prior to tissue collection, pain active genes were stimulated via one second static fiber (0.16g) to each hind paw (1x/30sec/5min). Overall design: Five minutes after touch stimulus to the hindpaws, brains were rapidly dissected, the bilateral anterior cingulate cortex was punch, and the punches for each animal were flash frozen. Frozen punches were used to generate single nuclei suspensions and the nuclei were processed for the 10x Genomics 3' gene expression assay (v3.1).