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Figure 3. Self-Regulative DV Patterning in Dorsal Half-Embryos Requires ADMP and Has the Opposite Transcriptional Regulation of BMP4 and Ventral BMP Antagonists. (A) Experimental design (n = 28 or more per experimental set). (B) Sox2 expression in uninjected siblings. (C) Control blastula dorsal halves self-regulate. (D) The ventral half forms belly-pieces lacking CNS. (E) In dorsal halves injected with Admp MO, much of the ectoderm becomes CNS. (F) Ventral half-embryos are unaffected by Admp MO. (G) RT-PCR analysis of whole embryos at stage 11 after injection of Chordin (1 and 5 ng) or BMP4 (0.5 and 2.5 ng) protein (in a volume of 40 nl) into the blastocoele cavity at stage 8. (H) Sox2 expression in control embryos at stage 20. Inset illustrates the dorsal and ventral centers marked by Chordin and Sizzled at stage 12. (I) Coinjection of Bambi and Sizzled MOs ventralizes the embryo. (J) A single dorsal injection of xAdmp mRNA (50 pg) into wild-type embryo decreases anterior CNS differentiation and increases Sizzled expression (inset). (K) Depletion of Bambi and Sizzled greatly sensitizes the embryo to the effects of Admp mRNA. (L) Model of regulatory interactions in the Xenopus embryonic field. Green lines indicate pro-BMP effects; red lines indicate anti-BMP effects |
