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Involvement of AP-2rep in morphogenesis of the axial mesoderm in Xenopus embryo

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Abstract

We have previously isolated a cDNA clone coding for Xenopus AP-2rep (activator protein-2 repressor), a member of the Krüppel-like factor family, and reported its expression pattern in developing Xenopus embryos. In the present study, the physiological function of AP-2rep in the morphogenetic movements of the dorsal mesoderm and ectoderm was investigated. Embryos injected with either AP-2rep or VP16repC (a dominant-negative mutant) into the dorsal marginal zone at the 4-cell stage exhibited abnormal morphology in dorsal structures. Both AP-2rep and VP16repC also inhibited the elongation of animal cap explants treated with activin without affecting the expression of differentiation markers. Whole-mount in situ hybridization analysis revealed that expression of brachyury and Wnt11 was greatly suppressed by injection of VP16repC or AP-2rep morpholino, but expression was restored by the simultaneous injection of wild-type AP-2rep RNA. Furthermore, the morphogenetic abnormality induced by injection of VP16repC or AP-2rep morpholino was restored by simultaneous injection of brachyury or Wnt11 mRNA. These results show that AP-2rep is involved in the morphogenesis of the mesoderm at the gastrula stage, via the brachyury and/or Wnt pathways.

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Acknowledgements

We thank Drs. R. Buettner and M. Asashima for supplying plasmids.

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Authors and Affiliations

  1. Graduate School of Science and Technology, Niigata University, Niigata, Japan

    Yoshinari Saito, Masanori Gotoh & Yasutaka Ujiie

  2. Department of Biology, Faculty of Science, Niigata University, 8050 Ikarashi-2, Nishi-ku, Niigata, 950-2181, Japan

    Yumi Izutsu & Mitsugu Maéno

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  1. Yoshinari Saito
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  2. Masanori Gotoh
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  3. Yasutaka Ujiie
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  4. Yumi Izutsu
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  5. Mitsugu Maéno
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Correspondence to Mitsugu Maéno.

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This work was partly supported by a grant-in-aid from The Ministry of Education, Science, and Culture of Japan.

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Supplement Fig. 1.

Differentiation of neural, muscle, and notochord tissues occurred in AP-2rep- or VP16repC-injected embryos. Embryos were injected with AP-2rep (1000 pg) or VP16repC (100 pg) into the dorsal and ventral marginal zone regions (all the blastomeres) at the 4-cell stage and cultured until stage 39. Expression of N-CAM (a-c), muscle actin (d-f), and keratan sulfate (g-i) was analyzed by whole-mount antibody staining (arrows positive signals stained by each antibody). All the embryos injected with either AP-2rep or VP16repC show positive staining with these markers (a, n=2; b, n=5; c, n=5; d, n=2; e, n=5; f, n=5; g, n=4; h, n=5; i, n=5) (PDF 70.32 KB)

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Saito, Y., Gotoh, M., Ujiie, Y. et al. Involvement of AP-2rep in morphogenesis of the axial mesoderm in Xenopus embryo. Cell Tissue Res 335, 357–369 (2009). https://doi.org/10.1007/s00441-008-0712-7

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