Figure 3

Figure 3

Detailed phenotypic and behavioral changes in lmo4a morphants. All panels represent lateral views of ears. (A–D) Otic morphology of lmo4a morphants at different stages displaying smaller otocysts and thicker otic epithelia compared to WT. (E,G) Hair cell number was reduced in lmo4a morphants, as determined from triple labeling with phalloidin (red, staining the cell framework and stereocilium), anti-myo6 (green, uniformly distributed throughout hair cell cytoplasm) and DAPI (blue, labeling the cell nucleus). (F,H) Embryos were double-labeled with phalloidin and anti-HuC (green, staining SAG), which revealed an evident reduction in the area of SAG in lmo4a morphants. (I,J) Statistical analysis of hair cell numbers of utricle and saccules between control and lmo4a morphants showed significant differences (*** p < 0.001, n = 20). (K) Statistical analysis of the area of SAG between control and lmo4a morphants showed significant differences (*** p < 0.001, n = 20). (L) Anterior, posterior and lateral protrusions were observed in control larvae at 48 hpf; (M) Anterior, posterior and lateral protrusions were fused while ventral protrusions grew out in the control group at 55 hpf. (N) The hubs of semicircular canals exhibited regular shapes in the control group at 72 hpf. (O) No protrusions were visible in lmo4a morphants at 48 hpf. (P) Only lateral protrusions were visible in lmo4a morphants at 55 hpf. (Q) Only lateral protrusions were visible in lmo4a morphants at 72 hpf.