Figure 2.

AAV2.CR2-Crry Reduces Brn3+ RGC Degeneration Onset and Progression

(A–F) Confocal images through the NFL and GCL of retinal whole mounts immunostained for Brn3, shown as maximum-intensity projections of 25–35 μm. Insets display RGC mosaics at high magnification. Retinas representative of 10-month-old naive (A–C) and AAV2.CR2-Crry-treated mice (D and E), with corresponding level of ON damage indicated as pre-degenerative, moderately degenerative, or severely degenerative. (F) Example of Brn3+ cell-sampling method. (G and K) Plot of the highest density of Brn3+ RGCs (mean number nuclei/mm²) for individual retinas at 10 (G) and 12 months (K) of age. (H and L) Stacked bar chart representing percent retinas at each level of RGC preservation per condition: pre-degenerative (mean Brn3+ RGC density/mm² >2,000, green), declining (1,000–2,000, yellow), and degenerative (<1,000, red). (I and M) Stacked bar chart representing percent retinas at each level of RGC depletion per condition: not depleted (0 degrees with less than 500 Brn3+ cells/mm², white), partial (1–359 degrees, gray), and complete (360 degrees, black). (J and N) Bar graph of mean depleted retinal area (degrees) per condition. (O and P) Confocal images of Thy1^CFP/+ RGCs from representative retinas of 10-month-old DBA/2J mice. Insets show their Brn3 co-expression at high magnification. Scale bars, 500 μm (A–F, O, and P) and 100 μm (insets). See Tables 3, 4, 5, and 6 for statistical analysis.