Figure 4.

Figure 4.

The mdka^mi5001 mutant retinas fail to regenerate cone photoreceptors. A, Immunocytochemistry for red/green cone photoreceptor marker, Zpr1. In WT retina, immature cone photoreceptors start to appear at 5 dpl, and regeneration largely completes by 14 dpl. In the mdka^mi5001 mutant, regenerating photoreceptors are absent at 5 dpl. At 7 dpl, very few cone photoreceptors appear. The number of cone photoreceptors is less at 14 dpl compared with WT. B, Immunocytochemistry for rod photoreceptor marker Zpr3 following lesion. In WT retina, regenerating rod photoreceptors appear by 7 dpl. In the mdka^mi5001 retinas, rod photoreceptors slowly regenerate by 28 dpl. C, Flat-mounted retinal preparation immunostained with ZO1 in unlesioned and 14 dpl. In unlesioned retina of both WT and mdka^mi5001, cone photoreceptors form a crystalline mosaic array in the planar apical surface of the retina (Livak and Schmittgen, 2001; Nagashima et al., 2017). Higher magnification of boxed region indicates the alignment of cones in the mosaic array (asterisks) with flattened cell boundaries (arrowheads). At 14 dpl in WT, cone photoreceptors regenerate (asterisks), although the crystalline mosaic array is not restored. In the mdka^mi5001 retina, cone profiles are instead replaced by irregularly shaped, expanded Müller glial apical processes (dotted line). D, Counts of ZO1-labeled cone photoreceptors at 14 and 28 dpl. Significantly fewer cones are regenerated in the mdka^mi5005 mutant (white) compared with WT (gray). n = 6. 14 dpl: p = 0.0051; 28 dpl: p = 0.0051, nonparametric Mann-Whitney-Wilcoxon. onl, Outer nuclear layer; inl, inner nuclear layer; gcl, ganglion cell layer. Scale bars: A, B, 30 μm; C, 10 μm. *p < 0.01.