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Fig. 1. Regulation of Pax3, Zic1 and Foxd3 expression in early Xenopus embryos. (A-C) The expression of Pax3 (A), Zic1 (B) and Foxd3 (C) at the early neurula stage as analyzed by whole-mount in situ hybridization. (D-F) Double-labeled in situ hybridization at the neurula stage (stage 15). (D) Pax3 (light blue) and Foxd3 (magenta); (E) Zic1 (light blue) and Foxd3 (magenta); (F) Pax3 (magenta) and Zic1 (light blue). (G-N) Effects of BMP and Wnt signals on Pax3, Zic1 and Foxd3 expression. Synthetic mRNAs of CA-BMPR (50 pg/cell; G-I), dnBMPR (200 pg/cell; J-L) or Wnt3a (10 pg of DNA/cell; M,N) were injected into two left animal blastomeres at the eight-cell stage. Whole-mount in situ hybridization was performed with Pax3 (G,J,M), Zic1 (H,K) and FoxD3 (I,L,N) probes. Arrows in J-N indicate ectopic expression of each marker gene; arrowheads in F indicate the Pac3+/Zic1+ area. |
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Fig. 2. Overexpression of Pax3 and Zic1 induces ectopic expression of neural crest markers. Synthetic mRNA was injected into two left animal blastomeres (A-L) or into the ventral animal blastomeres (M-P) at the eight-cell stage. Embryos were harvested at stage 15 and subjected to in situ hybridization with Foxd3 (A,D,G-M,O,P), Slug (B,E,N), Zic1 (C) and Pax3 (F) probes. Pax3 mRNA injection (50 pg/cell; A-C), Zic1 mRNA injection (50 pg/cell; D-F), Pax3-GR mRNA injection (100 pg/cell; G-I), Zic1-GR mRNA injection (100 pg/cell; J-L), and injection of both Pax3 and Zic1 mRNAs (50 pg/cell each; E-H) with lacZ mRNA (200 pg/cell). Dexamethasone (Dex; 10 μM) was added to culture solution at stage 10.5 (H,K) or stage 12 (I,L). Arrows in A,B,H,K,M and N indicate ectopic expression of each marker gene. |
