single-nucleus RNA-seq of enriched retinal ganglion cells (RGCs) with RGC-specific overexpression of hOprm1 under optic nerve crush injury condition

Through a comparative analysis of RGC subclasses exhibiting varying levels of resilience to cell death, based on cell-cell interaction analysis, we identified the Mu-opioid receptor, encoded by the Oprm1 gene, as a novel neuroprotective factor for pan-RGCs. Functionally and with the sequencing dataset deposited here, we validated Oprm1 as a neuroprotective factor for RGCs, by demonstrating that overexpression of Oprm1 in RGCs not only led to a markedly increased survival rate of RGCs following several different types of retinal injuries, but also significantly improved visually-guided perception behaviors. Overall design: Retinas from the vGlut2-Cre;LSL-Sun1GFP mice under 5-day ONC or 5-day ONC with hOprm1 overexpression were dissociated to expose cell nuclei. GFP+ nuclei of RGCs were enriched with anti-GFP MACS. Final concentration of 4000~5000 nuclei/uL loading onto the 10X Genomics Single-Nuclei 3' HT platform.