Expression of dfna5 in zebrafish embryos using whole-mount in situ hybridization. (A,B) 22 hpf embryo. Lateral (A) and dorsal view of the head region (B), showing expression in the intermediate cell mass (arrowhead in A), the olfactory placodes (arrows), ventral diencephalon and migrating neural crest (arrows in B). (C,D) 48 hpf embryo (C) and 55 hpf embryo (D) with expression in the developing semicircular canals of the ear (arrowheads). (E) 72 hpf embryo indicating low level of expression in the mature projections of the semicircular canals in the ear (arrowheads). (F) Dorsal view of the ear at 48 hpf with expression at the tip of the outgrowing projections of the semicircular canals (arrowheads). Scale bar: 300 µm in A,B; 100 µm in C-E; 40 µm in F.

In situ analysis of cartilage differentiation in dfna5 morphants. (A,D) Lateral view of col2a1 expression at 55 hpf. White arrows indicate the otic vesicle; white arrowheads indicate pharyngeal arches. Expression in the morphant otic vesicle (D) is unaffected compared with wild type (A). (B,C,E,F) ugdh expression in wild-type (B,C) and morphant (E,F) 55 hpf embryos. (B,E) Lateral view. (E) Expression in the developing morphant ear columns (indicated on wild type) and pharyngeal arches (white arrowhead) is reduced compared with wild-type (B). (C,F) Ventral view of expression in the developing neurocranium reveals no difference between wild-type and morphant embryos (arrows in C and F). Scale bar: 200 µm. c, column; e, eye; fb, fin bud; nc, neurocranium; ov, otic vesicle; pa, pharyngeal arches.