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

Nature volume 446, pages 79–82 (2007)Cite this article

Abstract

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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Figure 1: GSK-3β mutants have cleft palates.
Figure 2: GSK-3β mutants show bifid sternum and delayed sternal ossification.
Figure 3: Drug-dependent rescue of cleft palate GSK-3β FRB * /FRB * mice in utero.
Figure 4: Reversal of sternal defects in chemically sensitive GSK-3β embryos.

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Acknowledgements

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.

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Author notes

  1. Karen J. Liu & Joseph R. Arron

    Present address: 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.),

  2. Karen J. Liu and Joseph R. Arron: These authors contributed equally to this work.

Authors and Affiliations

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

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Liu, K., Arron, J., Stankunas, K. et al. Chemical rescue of cleft palate and midline defects in conditional GSK-3β mice. Nature 446, 79–82 (2007). https://doi.org/10.1038/nature05557

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