Fig. 3

dnah6 morpholino knockdown in zebrafish embryo cause laterality defects.

(A,B) dnah6 MO injected embryos exhibited curly tail and cardiac edema phenotype at 48 hours post fertilization (hpf). (C,D) dnah6 MO injected embryos at 48hpf exhibited heart (C) and gut (D) looping defects, including failure to loop (middle panels) and reversal looping (right panels). (E) In situ hybridization analysis revealed abnormal right sided (R), bilateral (B), and absent (Ab) spaw expression after dnah6 MO knockdown. (F,G) Heart/gut looping defects were observed in dnah6 MO-injected embryos, including normal (N), right sided (R), and straight (St) heart/gut looping phenotypes. (H). KV cilia were shorter (middle) in dnah6 AUG-MO but not dnah6 spl-MO injected embryos (p-value = 0.0017). (I). High-speed videomicroscopy showed reduction in KV motile cilia of dnah6 MO injected embryos (p = 1.6x10^-4 for 7.5 ng and p = 1.1x10^-5 for 10 ng dnah6 MO; two-tailed Student’s t-test). (J) Little bead movement (color tracing) was observed in dnah6 MO injected embryos. (K) KV flow was absent or reduced with dnah6 knockdown (Chi-square test, p-value = 0.0192).