Generation and characterisation of rdh12u533 mutant zebrafish. (A) Sanger sequencing traces showed a 7 bp deletion (c.17_23del) in the rdh12u533 mutant fish. The deleted bases are highlighted in blue on the wildtype trace. (B) RT-qPCR showed a significant reduction of rdh12 mRNA expression in the rdh12u533 fish at 5 dpf (** p ≤ 0.001 analysed by paired t-test). (C,D) Retinal sections from 12 mpf fish were stained with toluidine blue to assess retinal structure. (E,F) TUNEL assay revealed no cell death Immunohistochemistry staining was used to detect rhodopsin (red) (G,H), blue opsin (green) (I,J), and red/green opsin (green) (K,L). Sections were counterstained with DAPI (blue). Rhodopsin mislocalisation was observed in rdh12u533 fish. Scale bar = 50 µM. IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; IS, inner segment; OS, outer segment; RPE, retinal pigment epithelium.
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