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Fig. 5. Antisense Otx2 morpholino oligonucleotide rescues convergent extension and Xbra expression blocked by IGF-I and Otx2. Embryos at 4-cell stage were dorsaly-injected with IGF-I or Otx2-Myc mRNA alone or co-injected with Otx2 morpholino. They were either allowed to develop to various stages for phenotypes or fixed at stage 10.5 for in situ hybridization. (A) An uninjected embryo at stage 14 showing correct neural plate formation. (B) An Otx2-Myc-injected embryo with gastrulation defect and open blastopore. (C) Otx2 morpholino rescues blastopore closure and neural plate formation in Otx2-Myc-injected embryo. (D) Western blot showing that Otx2 morpholino specifically blocks translation of Otx2-Myc mRNA. (E) A control embryo at stage 38. (F) An Otx2-Myc-injected embryo at equivalent stage with dorsal development defect and shortened trunk and posterior region. (G) Otx2 morpholino rescues trunk and posterior development. (H–L) Otx2 morpholino rescues Xbra and Xwnt11 expression. (H) An uninjected stage 10.5 gastrula showing Xbra expression. (I) An IGF-I-injected early gastrula with absence of Xbra expression at injected site. (J) Rescue of Xbra expression by Otx2 morpholino in IGF-I-injected early gastrula. (K) An Otx2-Myc-injected early gastrula with a gap in Xbra expression in the marginal zone. (L) Rescue by Otx2 morpholino of Xbra expression. (M) An uninjected early gastrula showing Xwnt11 expression. (N) Dorsal injection of IGF-I mRNA inhibits Xwnt11 expression. (O) Rescue by Otx2 morpholino of Xwnt11 expression. |
