Figure 4.

Upregulation of glycolysis through ablation of PHD enhances cone survival in a preclinical Pde6β^H620Q/H620Q RP model

PHD deficiency improves rod and cone survival in the Pde6β^H620Q/H620Q mutant background. Experimental (PHD^−/−;Pde6β^H620Q/H620Q;Pde6γ^CreERT2/+) and control (PHD^FL/FL;Pde6β^H620Q/H620Q;Pde6γ^CreERT2/+) mice were treated with tamoxifen or sham solution for three consecutive days (P9, P10, and P11).

(A–C) Analysis of amplitudes of electroretinogram tracings comparing experimental and control mice. Mice were subjected to three types of serial ERG recordings: scotopic rod (A), maximal rod and cone (B), and photopic cone (C) at 6, 8, and 10 weeks postnatally. Experimental mice with precise PHD1,2,3 ablation in rod photoreceptors are shown in red, while the signal from control mice is in gray. The average ERG amplitude of both eyes per mouse was used for analysis. Error bars indicate SEM. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001; n = 5–6 in each group per time point.

(D) Representative H&E-stained central retinal sections from experimental and control mice at 4 and 6 weeks at a distance of 500 μm from the optic nerve head. Yellow bars indicate ONL thickness. Scale bars, 25 μm.

(E) Spider plot analysis of ONL thickness at 4 weeks. Error bars indicate SEM. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001; n = 6 per group.

(F) Spider plot analysis of ONL thickness at 6 weeks. Error bars indicate SEM. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001; n = 6 per group.

(G) Peanut agglutinin (PNA) staining of cones in 12-week-old PHD^fl/fl control and PHD^−/− (deficient) mice. The inset shows representative central retinal cones.

(H) Bar chart of cone cell counts, quantified by percent area of the total area measured in both groups in (G). Error bars indicate SEM. ∗∗p ≤ 0.01; n = 3 in each group.