Fig. 6: snRNA-seq of RGCs after Oprm1 overexpression.

a Clustering of RGC subclasses profiled by snRNA-seq as an integration of cells collected from sham conditions, 5 days post-ONC, and the Oprm1-overexpressed 5 days post-ONC conditions. b UMAP shows the expression patterns of endogenous Oprm1 (cyan) and the ectopic expression of human Oprm1 (pink) in RGC subclasses. c Representative micrographs of whole-mount retinal tissues show the expression patterns of Opn4 and Spp1 in Oprm1-expressing RGCs. The red fluorescent staining is the mCherry tag in the Oprm1-mCherry reporter mouse strain; the blue represents Opn4 immunostaining, and the green represents Spp1 immunostaining. Grey long arrows point to the low-level Opn4-expressing cells; short white arrows indicate the medium-level Opn4-expressing cells. The scale bar is 50 μm. d Quantification of the ratio of Opn1 and Spp1 expression patterns in Oprm1-expressing RGCs. Data are presented as mean values ± SEM. n = 5 biological replicates of mice. e Normalized cell proportion among RGC subclasses under three conditions. f Representative biological processes enriched in Oprm1-overexpressed five days post-ONC condition. Gene set enrichment analysis (GSEA) of Gene Ontology was adjusted using the Benjamini–Hochberg method. p-values for the GSEA test statistics are calculated by permutation (nPerm = 10,000). g Visual cliff test on the protective effect of Oprm1 overexpression against NMDA-induced vision impairment. Left: the equipment setup. Right: mean occurrences of mice selecting the shallow side (an indicator of acute vision) out of 10 trials. Each dot represents one mouse. Data are presented as mean values ± SEM. n = 5 mice for each independent experimental condition. Ordinary one-way ANOVA, Tukey’s multiple comparisons test, with a single pooled variance. h Pupil reflex test on naloxone effect in NMDA model. Left: representative images of the pupil in three conditions. Right: normalized pupil size in three conditions. Data are presented as mean values ± SEM. n = 6 or 7 mice for each independent experimental condition. Ordinary one-way ANOVA, Tukey’s multiple comparisons test, with a single pooled variance. Source data are provided as a Source Data file.