Abstract
The maternal genome greatly influences vertebrate embryogenesis before activation of zygotic transcription. Dorsoventral patterning is initiated by maternal factors1,2,3, but the molecular pathways involved are incompletely understood. In frogs and fish, localized zygotic domains are induced whereby cells either express dorsal or ventral genes1,2. Wnt/β-catenin signalling promotes expression of the earliest dorsal zygotic genes4. Among key zygotic ventralizing factors are the bone morphogenetic proteins (Bmps)5,6 and the Vent homeodomain family2,7,8,9,10; the latter act as repressors of dorsal organizer gene transcription. Here we show that Runx2, a transcription factor essential for bone formation, is an important maternal determinant of ventral zygotic genes in zebrafish. Depletion of maternal Runx2b type2 strongly dorsalizes embryos, due to loss of the earliest zygotic expression of vox, vent and ved, resulting in expansion of dorsal gene expression. To date, Runx2b is the only known regulator of vox, vent and ved at the onset of zygotic transcription; we show that this regulation is direct. Runx2 transcripts are processed in mature mouse oocytes11 and we show that murine Runx2 type2 can substitute for the zebrafish orthologue in its ventralizing function, suggesting that Runx2 may have an evolutionarily conserved role in axis formation.
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References
Heasman, J. Patterning the early Xenopus embryo. Development 133, 1205–1217 (2006).
Schier, A. F. & Talbot, W. S. Molecular genetics of axis formation in zebrafish. Annu. Rev. Genet. 39, 561–613 (2005).
Wang, H. & Dey, S. K. Roadmap to embryo implantation: clues from mouse models. Nature Rev. Genet. 7, 185–199 (2006).
Kelly, C., Chin, A. J., Leatherman, J. L., Kozlowski, D. J. & Weinberg, E. S. Maternally controlled (β)-catenin-mediated signaling is required for organizer formation in the zebrafish. Development 127, 3899–3911 (2000).
Kishimoto, Y., Lee, K. H., Zon, L., Hammerschmidt, M. & Schulte-Merker, S. The molecular nature of zebrafish swirl: BMP2 function is essential during early dorsoventral patterning. Development 124, 4457–4466 (1997).
Schmid, B. et al. Equivalent genetic roles for bmp7/snailhouse and bmp2b/swirl in dorsoventral pattern formation. Development 127, 957–967 (2000).
Imai, Y. et al. The homeobox genes vox and vent are redundant repressors of dorsal fates in zebrafish. Development 128, 2407–2420 (2001).
Gilardelli, C. N., Pozzoli, O., Sordino, P., Matassi, G. & Cotelli, F. Functional and hierarchical interactions among zebrafish vox/vent homeobox genes. Dev. Dyn. 230, 494–508 (2004).
Kawahara, A., Wilm, T., Solnica-Krezel, L. & Dawid, I. B. Antagonistic role of vega1 and bozozok/dharma homeobox genes in organizer formation. Proc. Natl Acad. Sci. USA 97, 12121–12126 (2000).
Shimizu, T. et al. A novel repressor-type homeobox gene, ved, is involved in dharma/bozozok-mediated dorsal organizer formation in zebrafish. Mech. Dev. 118, 125–138 (2002).
Hernandez-Gonzalez, I. et al. Gene expression profiles of cumulus cell oocyte complexes during ovulation reveal cumulus cells express neuronal and immune-related genes: does this expand their role in the ovulation process? Mol. Endocrinol. 20, 1300–1321 (2006).
Gore, A. V. et al. The zebrafish dorsal axis is apparent at the four-cell stage. Nature 438, 1030–1035 (2005).
Pei, W., Williams, P. H., Clark, M. D., Stemple, D. L. & Feldman, B. Environmental and genetic modifiers of squint penetrance during zebrafish embryogenesis. Dev. Biol. 308, 368–378 (2007).
Leung, T. et al. bozozok directly represses bmp2b transcription and mediates the earliest dorsoventral asymmetry of bmp2b expression in zebrafish. Development 130, 3639–3649 (2003).
Sidi, S., Goutel, C., Peyrieras, N. & Rosa, F. M. Maternal induction of ventral fate by zebrafish radar. Proc. Natl Acad. Sci. USA 100, 3315–3320 (2003).
Reim, G. & Brand, M. Maternal control of vertebrate dorsoventral axis formation and epiboly by the POU domain protein Spg/Pou2/Oct4. Development 133, 2757–2770 (2006).
Blyth, K., Cameron, E. R. & Neil, J. C. The RUNX genes: gain or loss of function in cancer. Nature Rev. Cancer 5, 376–387 (2005).
Ito, Y. & Miyazono, K. RUNX transcription factors as key targets of TGF-β superfamily signaling. Curr. Opin. Genet. Dev. 13, 43–47 (2003).
Flores, M. V., Lam, E. Y., Crosier, P. & Crosier, K. A hierarchy of Runx transcription factors modulate the onset of chondrogenesis in craniofacial endochondral bones in zebrafish. Dev. Dyn. 235, 3166–3176 (2006).
Flores, M. V. et al. Duplicate zebrafish runx2 orthologues are expressed in developing skeletal elements. Gene Expr. Patterns 4, 573–581 (2004).
Ramel, M. C. & Lekven, A. C. Repression of the vertebrate organizer by Wnt8 is mediated by Vent and Vox. Development 131, 3991–4000 (2004).
Ramel, M. C., Buckles, G. R., Baker, K. D. & Lekven, A. C. WNT8 and BMP2B co-regulate non-axial mesoderm patterning during zebrafish gastrulation. Dev. Biol. 287, 237–248 (2005).
Sander, V., Reversade, B. & De Robertis, E. M. The opposing homeobox genes Goosecoid and Vent1/2 self-regulate Xenopus patterning. EMBO J. 26, 2955–2965 (2007).
Ueta, C. et al. Skeletal malformations caused by overexpression of Cbfa1 or its dominant negative form in chondrocytes. J. Cell Biol. 153, 87–100 (2001).
Mullins, M. C. et al. Genes establishing dorsoventral pattern formation in the zebrafish embryo: the ventral specifying genes. Development 123, 81–93 (1996).
Ducy, P., Zhang, R., Geoffroy, V., Ridall, A. L. & Karsenty, G. Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89, 747–754 (1997).
Canalis, E., Deregowski, V., Pereira, R. C. & Gazzerro, E. Signals that determine the fate of osteoblastic cells. J. Endocrinol. Invest. 28, 3–7 (2005).
Canalis, E., Economides, A. N. & Gazzerro, E. Bone morphogenetic proteins, their antagonists, and the skeleton. Endocr. Rev. 24, 218–235 (2003).
Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B. & Schilling, T. F. Stages of embryonic development of the zebrafish. Dev. Dyn. 203, 253–310 (1995).
Nasevicius, A. & Ekker, S. C. Effective targeted gene 'knockdown' in zebrafish. Nature Genet. 26, 216–220 (2000).
Bjornson, C. R. et al. Eomesodermin is a localized maternal determinant required for endoderm induction in zebrafish. Dev. Cell. 9, 523–533 (2005).
Acknowledgements
We thank Alhad Mahagaonkar and Lisa Pullin for technical assistance. We are grateful to M. Hammerschmidt, M. Hibi, T. Komori, D. Kimelman and M. Mullins for sharing their reagents. We thank M. Hibi for his advice on the manuscript. The work was funded by grants from the Health Research Council of New Zealand and the Foundation for Research, Science and Technology, New Zealand. E.Y.L. was supported by Bright Futures Top Achievers Doctoral Scholarship.
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M. V. C. F. designed and performed the experimental work, analysed the data and wrote the manuscript. E. Y. L. performed some experiments and the statistical analysis. K. E. C. and P. S. C. planned the project and reviewed the manuscript.
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Supplementary figures S1, S2, S3, S4 and Supplementary table S1 (PDF 653 kb)
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Flores, M., Lam, E., Crosier, K. et al. Osteogenic transcription factor Runx2 is a maternal determinant of dorsoventral patterning in zebrafish. Nat Cell Biol 10, 346–352 (2008). https://doi.org/10.1038/ncb1697
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DOI: https://doi.org/10.1038/ncb1697
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