Maternal atrn mutation leads to epiboly delay. (A) CRISPR/Cas9-mediated deletion of atrn. Zebrafish atrn contains 29 exons, and gRNA was designed to target the second exon. Sequencing results of the target sites from several individual F2 embryos are shown below, including the wild-type form and two atrn mutant forms. (B) Maternal but not zygotic atrn mutation results in epiboly initiation and progression delays. Wild-type, Zatrn, Matrn and MZatrn mutant embryos were collected after fertilization at the same time and then imaged at indicated time points. (C, D) The quantitative data of epiboly initiation at 5 hpf and epiboly progression at 8 hpf in (B) are shown. (E) CE movements appear essentially normal in MZatrn mutant embryos. The mutant embryos were collected at both same time and morphologically comparable stages with wild-type embryos. dlx3b and ntla probes were used simultaneously for in situ hybridization. The ratios of embryos with representative pattern are indicated at the right corner of each picture. *, p<0.01. Scale bars: 100 μm in (B, E).
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