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Fig. 3 Ginger/10-G treatment during gastrulation promotes bmp2b/7a and Bmp target gene expression in zebrafish embryos.
(A) Treatment of late gastrulae with ginger at 15 or 20 μg/ml induces the mercedes mutant-like phenotype (partial duplication of the tail fin) at 1 dpf in 8% or 10% of the treated embryos, respectively. Thus, the zebrafish embryos exposed to ginger extract mimic the phenotype of the ogon mutant, which has a mutation in sizzled, a bmp suppressor gene, at 1 dpf. (B) bmp7a expression was strongly increased and extended to the entire blastoderm at 60% epiboly, following short-term exposure to ginger (5 μg/ml) or 10-G (1 μg/ml) from sphere (4 hpf) to 60% epiboly (7 hpf) stages. (C) Up-regulation and extension of the expression domain were observed for bmp2b at 60% epiboly. (D–E) Accordingly, BMP target genes were up-regulated after ginger/10G treatment from the sphere stage (4 hpf) to 7 hpf, as illustrated by enhanced eve1 extended towards the dorsal side (arrow heads), a ventral mesoderm marker (D), and gata2, a non-neural ectoderm marker (E), in zebrafish embryos at 60% epiboly. Pictures on left panels show gastrulae, dorsal side to the right (B–E) and statistics tables (right panels) are representative of three independent experiments. N = number of embryos per group. Scale bars = 250 μm.