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. 2014 Dec 8;111(51):E5574–E5583. doi: 10.1073/pnas.1414162111

Fig. 6.

Fig. 6.

LiGluR expression in RGCs restores light responses in the rcd1 canine retina in vitro. (A) Viral DNA expression cassette. LiGluR expression is driven by CAG promoter. (B) Schematic of degenerated canine retina with targeted RGCs in green. (C and D) Confocal images of LiGluR expression in canine retina 8 wk after intravitreal injection of AAV2/2 (4YF)-CAG-LiGluR. (Scale bars: 20 μm.) (C) Whole-mount view of LiGluR expression in RGCs of rcd1 canine retina. (D) Cross-section of LiGluR expression in RGCs of wt canine retina. (E–J) MEA recordings (light intensity: 75 mW/cm2 or 1.7 × 1017 photons per cm−2⋅s−1 if not specified otherwise) of rcd1 mutant canine retinas. MEA recording from an rcd1 mutant dog expressing LiGluR (n = 160 cells) before (E) and after (F) in vitro application of MAG0460. (G) Comparison of the firing rate averaged over a 3-s flash of light (500-ms bins) on three LiGluR-expressing rcd1 retinas in the dark vs. light (dashed line, 6.8 mW/cm2 or 1.5 × 1016 photons per cm−2⋅s−1; solid lines, 75 mW/cm2 1.7 × 1017 photons per cm−2⋅s−1) before (n = 2 retinas, 193 cells, gray) and after (n = 3 retinas, 291 cells, green) in vitro application of MAG0460, with population means (black and blue, respectively). (H) Dynamic range of rcd1 retina treated with LiGluR-MAG0460 at lower light conditions (n = 80 ± 5 cells). Data are mean ± 2 SEM binned at 100 ms. (I) Averaged normalized response to stimulation with five light intensities (0.43, 0.85, 1.7, 3.4, and 6.8 mW/cm2 or 9.7 × 1014, 1.9 × 1015, 3.8 × 1015, 7.7 × 1015, and 1.5 × 1016 photons per cm−2⋅s−1, respectively) shown in H with individual responses shown in gray, average response over all intensities shown in black, and the response inactivation time shown in blue (ΔToff). (J) MEA recording from an rcd1 retina treated with LiGluR and MAG0460 (n = 77 cells) stimulated at low intensity (0.85 mW/cm2 or 1.9 × 1015 photons per cm−2⋅s−1) at 4 Hz (50-ms duration, 200-ms ISI) binned at 10 ms. (Inset) Average of 400 individual responses (5-ms bins) with a single exponential fit (blue trace) and time constant (τoff ∼ 28 ms) for MAG0460 relaxation from peak response to baseline.