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. 2022 Mar 11;13:1282. doi: 10.1038/s41467-022-28982-6

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, c Representative images of semi-thin plastic sections stained with Richardson solution in wildtype (top) and mzbbs1^−/− (bottom) retina at 7 dpf (a) and 10 dpf (c) showing slight morphological abnormalities at 7 dpf and more severe changes at 10 dpf with shorter and misshapen OSs. b, d DiO-staining highlighting OSs reveals progressive morphological alterations of the OSs between 7 dpf (b) and 10 dpf (d). e TEM reveals bulky OSs (*asterisk) with abnormal membrane disc stacking in mutant retina at 7 dpf. Disc membranes in mutants (middle & right image) are less compact in places or stacked vertically compared to controls (red arrowheads). Membrane-filled swirls are frequently observed in the RPE (yellow arrows) at that stage (middle & right image are two images of mzbbs1^−/− retinae showing various alterations seen in the mutants). f At 8dpf, UV-Opsin (green) localizes normally to the OS in mzbbs1^−/− mutants and controls, despite abnormal ultrastructure and shortened OS revealed by the membrane dye BODIPY (magenta). g Top panel shows bar plots of the maximum b-wave amplitude by electroretinography at 10 dpf. Reduced b-wave intensity at various light intensities demonstrates severely decreased response to light. Unpaired two-tailed multiple T-test; Sig: ns = FDR(q value) = 0.089, **** = FDR(q value) < 0.0001; Sample size (n = 13 WT, n = 8 Mut larvae), Error bars show standard deviation around the mean. The bottom panel shows the average response curve after a 100 ms light flash at log-2 intensity (Light On/Off line below the ERG curve) for mutant (grey) compared to control (black). Scale Bars: (a,c) 12 µm, (b, d, f) 50 µm, (e) 5 µm. Abbreviations: OS outer segment, ERG electroretinography, INL inner nuclear layer, M mitochondria, N nucleus, ONL outer nuclear layer, OPL outer plexiform layer, P pigment, TEM transmission electron microscopy.