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Posterior malformations in Dact1 mutant mice arise through misregulated Vangl2 at the primitive streak

Nature Genetics volume 41, pages 977–985 (2009)Cite this article

Abstract

Mice homozygous for mutations in Dact1 (also called Dapper or Frodo) phenocopy human malformations involving the spine, genitourinary system and distal digestive tract. We traced this phenotype to disrupted germ-layer morphogenesis at the primitive streak. Notably, heterozygous mutation of Vangl2, a transmembrane component of the planar cell polarity (PCP) pathway, rescued recessive Dact1 phenotypes, whereas loss of Dact1 reciprocally rescued semidominant Vangl2 phenotypes. We show that Dact1, an intracellular protein, forms a complex with Vangl2. In Dact1 mutants, Vangl2 was increased at the primitive streak, where cells ordinarily undergo an epithelial–mesenchymal transition. This is associated with abnormal E-cadherin distribution and changes in biochemical measures of the PCP pathway. We conclude that Dact1 contributes to morphogenesis at the primitive streak by regulating Vangl2 upstream of cell adhesion and the PCP pathway.

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Figure 1: Birth phenotypes in Dact1 mutants compared to wild type.
Figure 2: Dact1 mutant embryonic phenotypes.
Figure 3: Biochemical correlates of PCP signaling are specifically affected at early phenotypic stages.
Figure 4: Mutations in Dact1 and Vangl2 mutually rescue.
Figure 5: The Dact1 and Vangl2 proteins associate independently of Dvl.
Figure 6: Dact1 and Vangl2 functionally interact at the primitive streak.

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Acknowledgements

We thank all members of the Cheyette laboratory for thoughtful input and assistance with mouse husbandry and genetics. E.P. Fox provided technical expertise for creation of the Dact1 targeting construct, starting from the pGKneoF2L2DTA2 vector donated by P. Soriano (Fred Hutchinson Cancer Research Center). Embryonic stem cell manipulation and chimera production were accomplished at the University of Washington Transgenic Resources Program. U. Grieshammer provided expert assistance with neonatal genitourinary phenotyping. Confocal microscopy was accomplished in the Nikon Imaging Center of the California Institute for Quantitative Biosciences at the University of California, San Francisco, with advice from K. Thorn. S. Dymecki (Harvard Medical School) provided the Tg(ACTFLPe)9205Dym transgenic mouse line, A. McMahon (Harvard University) the Wnt3atm1Amc mutant mouse line and S. Piccolo (University of Padua) the Tg(BAT-LacZ)3Picc line. T. Gridley (The Jackson Laboratory) provided probes for Uncx and Dll1. M. Montcouquiol (INSERM/Université Bourdeaux II) provided affinity-purified Vangl2 antibody. A. Wynshaw-Boris provided helpful comments on the final manuscript. B.N.R.C. gratefully acknowledges support from US National Institutes of Health K08MH001750 and R01HD055300, the Sandler Foundation, the UCSF Academic Senate Committee on Research and the UCSF Center for Neurobiology and Psychiatry. R.S. is a recipient of a predoctoral fellowship from the California Institute of Regenerative Medicine. D.A.C.F. received support from the Medical Scientist Training Program at Washington University St. Louis.

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Author notes

  1. Rowena Suriben, Saul Kivimäe and Daniel A C Fisher: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Psychiatry, University of California, San Francisco, San Francisco, California, USA

    Rowena Suriben, Saul Kivimäe, Daniel A C Fisher & Benjamin N R Cheyette

  2. Graduate Program in Developmental Biology, University of California, San Francisco, San Francisco, California, USA

    Rowena Suriben & Benjamin N R Cheyette

  3. and Department of Pharmacology, Howard Hughes Medical Institute, Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, Washington, USA

    Randall T Moon

  4. Graduate Program in Neuroscience, University of California, San Francisco, San Francisco, California, USA

    Benjamin N R Cheyette

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Contributions

R.S. contributed gene expression, QPCR (Wnt signaling targets), most embryological analyses, confocal microscopy and genetics. S.K. contributed antigen synthesis, antibody characterization, protein blots, kinase assays, coIPs, pulldowns and associated plasmid constructs. D.A.C.F. contributed skeletal and somitogenic analyses, BAT-gal WISH and genetics including the initial Vangl2Lp cross. B.N.R.C. designed the Dact1 targeting construct in the laboratory of R.T.M. at the University of Washington. The experiments of R.S., S.K. and D.A.C.F. were performed at UCSF in the laboratory of B.N.R.C., who supervised this research and wrote the text with feedback from all authors.

Corresponding author

Correspondence to Benjamin N R Cheyette.

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Competing interests

The Dact1 antibody described in this article, which was made in the laboratory of B.N.R.C., is being licensed by the University of California, San Francisco, to AbD Serotec for distribution.

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Suriben, R., Kivimäe, S., Fisher, D. et al. Posterior malformations in Dact1 mutant mice arise through misregulated Vangl2 at the primitive streak. Nat Genet 41, 977–985 (2009). https://doi.org/10.1038/ng.435

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