The expression of cadm3 and phenotypes of cadm3-disrupted retinae. (A) Quantification of expression (FPKM) of Cadm family genes (cadm1a, cadm1b, cadm2a, cadm2b, cadm3, and cadm4) in WT donor and p15⁺ host RPCs. (B) Design of BAC plasmid of cadm3:mNeongreen. (C)cadm3:mNeongreen transgenic fish line showing the expression patterns of cadm3 at 32, 36, 48, and 72 hpf. (D) Image showing gel electrophoresis of DNA fragments resulting from genomic editing by Cas9/cadm3 sgRNAs (region 1 covering sg-1 and sg-2; region 2 covering sg-3 and sg-4). (E) Representative images showing neurogenesis wave and retinal lamination in scramble-sg and cadm3-disrupted retinae by Tg(atoh7:gapRFP:: ptf1α:GFP) fish line, in which RGCs and PRs are labeled by Tg(atoh7:gapRFP), and ACs and HCs are labeled by Tg(ptf1α:GFP).(F) F-actin distribution visualized by GFP-Utrch mRNA injection in scramble-sg (top) and cadm3-disrupted (down) retinae. Zoom-in images of retinal areas (dashed yellow rectangles) are shown in the right panels. Areas within white dashed contour lines are retinae in C, E, and F. (G and H) Representative cases (G) and quantification of cell-cycle length (H) of single 12-hpf scramble-sg (top, n = 16) and cadm3-disrupted (bottom, n = 15) RPCs. Yellow arrowheads show RPCs at the M phase. The area of dashed white circles is the optic vesicle, and the area between two dashed white lines is the retina. Data in A and H represent mean ± SEM. Scale bars, 10 µm. Source data are available for this figure: SourceData FS3.