Fig. 1

Spatiotemporal expression and mutations in zebrafish dchs genes leading to pleiotropic defects during embryogenesis. (A) qRT-PCR analysis of the expression of all three zebrafish dchs genes at maternal and zygotic stages normalized to gapdh transcripts. (B) Whole-mount in situ hybridization (WISH) of dchs1b in WT embryos at four-cell, shield, 90% epiboly and 24-hpf stages. (C) Sanger sequencing trace for dchs2 A-to-T mutations. (D) Schematic of Dchs protein, with mutations denoted by asterisks. (E) qRT-PCR analysis of dchs1b expression in MZdchs1b relative to WT embryos at maternal and zygotic stages. (F) Bright-field images of WT, MZdchs1b, MZdchs2^stl1/stl1 and MZdchs1b^fh275/fh275;MZdchs2^stl1/stl1 time-matched embryos at 1, 6 and 10hpf. Red boxes indicate distortions in the yolk cell. Large arrowheads denote yolk masses in the blastoderm. Small red arrowheads indicate anterior (A); small blue arrowheads indicate posterior (P). (G) ntl WISH for stage-matched WT, MZdchs1b and MZdchs2^stl1/stl1 embryos at 70% epiboly. Inset depicts time-matched MZdchs1b embryo with a gap in the ntl expression domain. (H) Quantification of axial mesoderm length in WT (n=111), MZdchs1b (n=414) and MZdchs2^stl1/stl1 (n=486) embryos. (I) hgg1 and dlx3 WISH analysis of WT, MZdchs1b and MZdchs2^stl1/stl1 stage-matched embryos at two-somite stage (12hpf). (J) Quantification of the mediolateral width of dlx3 domain for WT (n=6), MZdchs1b (n=27) and MZdchs2^stl1/stl1 (n=18), shown by black line.