Manipulation of Wnt affects zebrafish jaw joint morphology. (A,A′) 5 dpf DMSO control (A) and 5 dpf 20 µM IWR-1-treated (from 3-5 dpf) (A′) zebrafish jaw joint morphology. Left: Tg(Col2a1aBAC:mcherry) transgenic zebrafish line marks the cartilage of the jaw joint. Middle: 3D volume rendering of jaw joint. Right: outlines of four representative jaw joints. A, anterior; P, posterior; M, medial; L, lateral. White lines indicate interzone interval measurements between MC and PQ; yellow line indicates overlapping interval between MC and PQ. (B,B′) Interzone intervals (µm) between the MC and PQ on the medial (B) and lateral (B′) regions of the jaw joint in 5 dpf DMSO and IWR-1-treated zebrafish. Negative values represent an overlap of MC/PQ elements (n=42 and 45 joints). Two-tailed Student's t-tests were performed for B,B′. (C,C′) 5 dpf control-injected (C) and Wnt16 morpholino (MO)-injected (C′) zebrafish jaw joint morphology. Left: immunohistochemical stain of the jaw joint region. Middle: 3D volume rendering of the jaw joint. Right: outlines of four representative jaw joints. (D,D′) Interzone intervals (µm) between the MC and PQ on the medial (D) and lateral (D′) regions of the jaw joint in 5 dpf control-injected and Wnt16 MO-injected zebrafish. Negative values represent an overlap of MC/PQ elements (n=8, 11, 6 and 8 joints). One-way ANOVA calculations were performed (D,D′). ns, not significant, *P≤0.05, ***P≤0.001. Data are mean and 95% confidence interval. (E) 3D volume rendering of 5 dpf injected CRISPR/Cas9 mosaic wnt16 knockout Tg(Col2a1aBAC:mcherry) larvae (1,2) (n=12 animals). Image was enlarged and rotated to best show the jaw joint (1′,2′). Right: outlines of four representative jaw joints.
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