Letter | Published:

Chemical rescue of cleft palate and midline defects in conditional GSK-3β mice

Nature volume 446, pages 7982 (01 March 2007) | Download Citation


Glycogen synthase kinase-3β (GSK-3β) has integral roles in a variety of biological processes, including development, diabetes, and the progression of Alzheimer’s disease1,2,3,4. As such, a thorough understanding of GSK-3β function will have a broad impact on human biology and therapeutics. Because GSK-3β interacts with many different pathways, its specific developmental roles remain unclear5. We have discovered a genetic requirement for GSK-3β in midline development. Homozygous null mice display cleft palate, incomplete fusion of the ribs at the midline and bifid sternum as well as delayed sternal ossification. Using a chemically regulated allele of GSK-3β (ref. 6), we have defined requirements for GSK-3β activity during discrete temporal windows in palatogenesis and skeletogenesis. The rapamycin-dependent allele of GSK-3β produces GSK-3β fused to a tag, FRB* (FKBP/rapamycin binding), resulting in a rapidly destabilized chimaeric protein. In the absence of drug, GSK-3βFRB*/FRB* mutants appear phenotypically identical to GSK-3β-/- mutants. In the presence of drug, GSK-3βFRB* is rapidly stabilized, restoring protein levels and activity6. Using this system, mutant phenotypes were rescued by restoring endogenous GSK-3β activity during two distinct periods in gestation. This technology provides a powerful tool for defining windows of protein function during development.

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  1. 1.

    & The renaissance of GSK3. Nature Rev. Mol. Cell Biol. 2, 769–776 (2001)

  2. 2.

    & GSK3 takes centre stage more than 20 years after its discovery. Biochem. J. 359, 1–16 (2001)

  3. 3.

    & GSK-3: tricks of the trade for a multi-tasking kinase. J. Cell Sci. 116, 1175–1186 (2003)

  4. 4.

    & The glamour and gloom of glycogen synthase kinase-3. Trends Biochem. Sci. 29, 95–102 (2004)

  5. 5.

    et al. Requirement for glycogen synthase kinase-3β in cell survival and NF-κB activation. Nature 406, 86–90 (2000)

  6. 6.

    et al. Conditional protein alleles using knockin mice and a chemical inducer of dimerization. Mol. Cell 12, 1615–1624 (2003)

  7. 7.

    & Cleft palate: players, pathways, and pursuits. J. Clin. Invest. 113, 1676–1678 (2004)

  8. 8.

    , , & Inhibition of Hedgehog signaling by direct binding of cyclopamine to Smoothened. Genes Dev. 16, 2743–2748 (2002)

  9. 9.

    , , , & Signals transduced by Ca2+/calcineurin and NFATc3/c4 pattern the developing vasculature. Cell 105, 863–875 (2001)

  10. 10.

    et al. Neurotrophins and netrins require calcineurin/NFAT signaling to stimulate outgrowth of embryonic axons. Cell 113, 657–670 (2003)

  11. 11.

    et al. Chemical suppression of a genetic mutation in a zebrafish model of aortic coarctation. Nature Biotechnol. 22, 595–599 (2004)

  12. 12.

    , , & Small molecule developmental screens reveal the logic and timing of vertebrate development. Proc. Natl Acad. Sci. USA 97, 12965–12969 (2000)

  13. 13.

    et al. A small-molecule agonist of the Wnt signaling pathway. Angew. Chem. Int. Ed. 44, 1987–1990 (2005)

  14. 14.

    & Mesenchymal influences on epithelial differentiation in developing systems. J. Cell Sci. 10 , (suppl.)195–230 (1988)

  15. 15.

    & Reaching a genetic and molecular understanding of skeletal development. Dev. Cell 2, 389–406 (2002)

  16. 16.

    , & Regulation of bone mass by Wnt signaling. J. Clin. Invest. 116, 1202–1209 (2006)

  17. 17.

    Studies on the morphogenesis of the mouse sternum. I. Normal embryonic development. J. Anat. 86, 373–386 (1952)

  18. 18.

    et al. Conditional inactivation of Tgfbr2 in cranial neural crest causes cleft palate and calvaria defects. Development 130, 5269–5280 (2003)

  19. 19.

    et al. FRAP/mTOR is required for proliferation and patterning during embryonic development in the mouse. Proc. Natl Acad. Sci. USA 98, 13796–13801 (2001)

  20. 20.

    , , & Transamniotic fetal feeding. II. A model of intrauterine growth retardation using the relationship of “natural runting” to uterine position. J. Pediatr. Surg. 22, 816–819 (1987)

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We thank M. S. Dionne for critical reading of the manuscript; M. S. Dionne, M. M. Winslow, J. E. Gestwicki, J. H. Bayle, S. C. Kao and members of the Longaker and Crabtree laboratories for invaluable discussions; and J. Woodgett for the gift of GSK-3β knockout mice. K.J.L and M.T.L. are supported by the NIH, M.T.L. is also supported by the Oak Foundation and J.R.A. is a fellow of the Berry Foundation.

Author information

Author notes

    • Karen J. Liu
    •  & Joseph R. Arron

    Present addresses: Department of Craniofacial Development, King’s College London, London SE1 9RT, UK (K.J.L.); Department of Immunology Diagnostics, Genentech, 1 DNA Way, South San Francisco, California 94080, USA (J.R.A.)

    • Karen J. Liu
    •  & Joseph R. Arron

    These authors contributed equally to this work.


  1. The Department of Pathology,

    • Karen J. Liu
    • , Joseph R. Arron
    •  & Gerald R. Crabtree
  2. The Stanford Institute for Stem Cell Biology and Regenerative Medicine,

    • Karen J. Liu
    •  & Michael T. Longaker
  3. Department of Developmental Biology,

    • Kryn Stankunas
    •  & Gerald R. Crabtree
  4. Howard Hughes Medical Institute, and the

    • Gerald R. Crabtree
  5. Division of Plastic and Reconstructive Surgery, Department of Surgery, Stanford University School of Medicine, Palo Alto, California 94305, USA.

    • Michael T. Longaker


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Competing interests

Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Corresponding authors

Correspondence to Karen J. Liu or Michael T. Longaker.

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    Supplementary Information

    This file contains Supplementary Methods and Supplementary Figures S1-S2 with legends.

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