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Fig. 7. Maternal Vangl2 and aPKC depletion disrupts the pattern of expression of specific zygotic genes. (A) Relative expression levels of Siamois, Xnr3 and Foxi1e mRNAs in control (Uninj) and Vangl2-depleted (VG–) embryos at the late blastula (st9) and early gastrula stages (st10) determined by real-time RT-PCR (mean±s.d.). (B) Phenotype of embryos derived from sibling control (Uninjected) and Vangl2-depleted oocytes (VG AS5, 4 and 6 ng oligonucleotide injected; VG AS4, 4, 6 and 8 ng oligonucleotide injected) at the early tailbud stage. (C) Relative expression levels of Siamois, Xnr3 and Foxi1e in control (Uninj) and aPKC-depleted embryos (AS5, 5 ng; AS6, 10 ng) determined by real-time RT-PCR at the late blastula and early gastrula stages. The experiment was repeated with a similar result. (D) Phenotype of embryos derived from sibling control (Uninjected) and aPKC-depleted oocytes (AS5, 3, 4 and 5 ng; AS6, 5 and 10 ng) at the early tailbud stage. (E) TOPflash reporter activation after injection into two dorsal vegetal cells of 8-cell stage control embryos compared with sibling Vangl2-depleted (left-hand graph) and aPKC-depleted (right-hand graph) embryos frozen at the late blastula stage, showing the reduction in canonical Wnt signaling activity (mean±s.d.). (F) Percentage of embryos with different phenotypes derived from Vangl2-depleted (AS4 and AS5), aPKC-depleted (AS5 and AS6) and sibling control at the tailbud stage. (G) Histological section of whole-mount stained, in situ hybridization of sibling control (upper) and aPKC-depleted (lower) early gastrulae using a Foxi1e probe. Arrow indicates ectopic expression of Foxi1e in the vegetal mass. (G′) Whole-mount in situ hybridization of sibling control (upper) and Vangl2-depleted (lower) early gastrulae using a Sox17a probe. (H) Relative expression levels of Xbra, Wnt11, Sox17a and Foxi1e in equatorial zones of control (Uninj.), Vangl2-depleted (VG–) and aPKC-depleted (aPKC–) embryos dissected at the late blastula stage and assayed by real-time RT-PCR (mean±s.d.). |