FIGURE 6.

RGC regeneration in Brn3b^AP/AP mice. (A) Schematic of the AAV constructs and infection procedure to regenerate RGCs in Brn3b^AP/AP mice at 1 month of age. (B,C) Flat-mounts of Brn3b^AP/AP retinas treated with GFAP-tdTomato AAVs were double-immunolabeled with anti-tdTomato and anti-Tuj1 antibodies. (D–G) Flat-mounts of Brn3b^AP/AP retinas treated with GFAP-Math5-Brn3b-tdTomato AAVs were double-immunolabeled with anti-tdTomato and anti-Tuj1 antibodies. Shown in (E–G) are representative images from the central, intermediate and peripheral retinas, respectively. (H) The optic nerves from Brn3b^AP/AP mice treated with GFAP-Math5-Brn3b-tdTomato AAVs were immunoreactive for tdTomato whereas those from Brn3b^AP/AP mice treated with GFAP-tdTomato AAVs were not. (I–K) The optic nerves, optic chiasms and optic tracts from Brn3b^AP/AP mice treated with GFAP-Math5-Brn3b-tdTomato AAVs were immunoreactive for tdTomato. Shown in (J) is a higher magnification view of the region outlined in (I). (L) Flat-mounts of central, intermediate and peripheral Brn3b^AP/AP retinas treated with GFAP-tdTomato or GFAP-Math5-Brn3b-tdTomato AAVs were immunostained with an anti-Rbpms antibody. (M) Quantification of Rbpms+ cells in central, intermediate and peripheral Brn3b^AP/AP retinas treated with GFAP-tdTomato or GFAP-Math5-Brn3b-tdTomato AAVs. Data are presented as mean ± SD (n = 4). Asterisks indicate significance in unpaired two-tailed Student’s t-test: *p < 0.005, **p < 0.001, ***p < 0.0005. AB, axon bundle; EF, MG endfoot; OC, optic chiasm; OD, optic disk; ON, optic nerve; OT, optic tract; RGC, retinal ganglion cell. Scale bars = 114 μm (I), 80 μm (B,D,H), 40 μm (J,K), 20 μm (C,E–G,L).