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Down-regulation of rod-specific genes and increased number of green cones in adult <italic toggle='yes'>nrl</italic>-KO zebrafish.(A) The protein levels of rod-specific (rho, gnat1, and gnb1) and cone-specific (gnat2 and gnb3) genes in WT and nrl-KO retinas from 14 dpf to 3 mpf were evaluated using western blotting. Tubulin was used as a loading control. The asterisk indicates a non-specific band. (B) Quantitative analysis of the protein levels of rod- and cone-specific genes based on at least three independent experiments. WT samples, blue color; nrl-KO samples, red color. Data from different ages are arranged from left to right (from 14 dpf to 3 mpf) and indicated by different fill patterns. The data are shown as mean with SD (n = 3). *, p < 0.05. **, p < 0.01. (C) The mRNA levels of the rod and cone opsins in the WT and nrl-KO retinas at 2 mpf and 5 mpf were measured using qPCR. The data are shown as mean with SD (n = 3). ns, non-significant; *, p < 0.05; **, p < 0.01. (D) Detection of green-cones on retinal sections of WT and nrl-KO zebrafish at 2 mpf, 3 mpf, and 13 mpf by immunostaining using the anti-Opn1mw antibody. The dorsal retinal regions are shown. White arrows indicate the mislocalized green-cone outer segments. GCOS, green-cone outer segments; CN, cone nuclear layer; RN, rod nuclear layer; INL, inner nuclear layer. Scale bars: 50 μm.
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