Fig. 4: ipRGCs are the main subtype reforming functional synapses and ipRGCs axonal regeneration partially restores PLR.

A Quantification of the percentages of Melanopsin+ ipRGCs, SMI32+ αRGCs, and other RGC types in PRV-GFP traced cells from the respective groups. Data presents mean ± SEM. n = 3 mice. Statistical significance: p = 0.0216 (Mela+ RGCs), p > 0.999 (SMI32+ RGCs), p = 0.0422 (Other RGCs). ANOVA followed by Bonferroni test, two-sided. B Fluorescence images showing PRV-traced cells (green) co-localized with ipRGC marker Melanopsin (red) (left panel) and αRGC marker SMI32 (red) (right panel) in the 6 m Regen. group. Scale bar: 50 µm. C Schematic diagram illustrating the experimental design for (D–F). D Quantification of percentages of pupil constrictions during PLR tests in different groups at different times post-injury. Data presents mean ± SEM. n = 5 (Injury Ctrl.) and n = 6 (ipRGC Regen.). Statistical significance: p ≤ 0.0001, ANOVA followed by Bonferroni test, two-sided. E Fluorescence images showing CTB-labeled RGC axons (green) and GFAP-labeled lesion site (red) in LGN and OPN from different groups. The dash lines outline the OPN regions. Scale bar: 200 µm. F Quantification of CTB-FITC intensities from labeled RGC axons at different distances from the lesion site. Blue asterisks indicate the comparison between 6 m ipRGC Regen. group and 12 m Injury Ctrl. group, while red asterisks represent the comparison between the 12 m ipRGC Regen. group and 12 m Injury Ctrl. group. Data presents mean ± SEM. n = 5 (12 m Injury Ctrl. and 6 m ipRGC Regen. group) and n = 6 (12 m ipRGC Regen.). Statistical significance: Red asterisks: p < 0.0001 (100 µm), p = 0.0412 (300 µm), Blue asterisks: p < 0.0001 (100 µm), p = 0.0034 (200 µm). ANOVA followed by Dunnett’s test, two-sided. ns, not significant, p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 Source data are provided as a Source Data file.