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Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins

Nature volume 433, pages 317–322 (2005)Cite this article

Abstract

Select members of the Wnt family of secreted glycoproteins have been implicated in inducing the myogenic determinant genes Pax3, MyoD and Myf5 during mammalian embryogenesis1,2, but the mechanism of induction has not been defined. We describe an unexpected role for protein kinase A (PKA) signalling via CREB in this induction. Using a combination of in vitro explant assays, mutant analysis and gene delivery into mouse embryos cultured ex vivo, we demonstrate that adenylyl cyclase signalling via PKA and its target transcription factor CREB are required for Wnt-directed myogenic gene expression. Wnt proteins can also stimulate CREB-mediated transcription, providing evidence for a Wnt signalling pathway involving PKA and CREB. Our findings raise the possibility that PKA/CREB signalling may also contribute to other Wnt-regulated processes in embryonic patterning, stem cell renewal and cancer3.

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Figure 1: P-CREB staining patterns coincide with myogenic induction.
Figure 2: CREB mutants have myogenic defects.
Figure 3: Functional elimination of the CREB family inhibits myogenesis in vivo.
Figure 4: CREB acts downstream of Wnt signalling.
Figure 5: G proteins, AC, cAMP and PKA mediate Wnt-induced myogenesis.

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Acknowledgements

We thank D. Koshland, Y. Zheng, A. Fire, L. Buttitta and M. Tessier-Lavigne for critical reading of this manuscript and C. Pratt for graphics assistance. D.D.G. is an investigator of the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

  1. Department of Embryology, Carnegie Institution of Washington, 115 West University Parkway, Baltimore, Maryland, 21210, USA

    Alice E. Chen & Chen-Ming Fan

  2. Department of Neuroscience, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA

    David D. Ginty

Authors
  1. Alice E. Chen
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  3. Chen-Ming Fan
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Corresponding author

Correspondence to Chen-Ming Fan.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure Legends

Text to accompany the below Supplementary Figures. (RTF 14 kb)

Supplementary Figure S1

Immunohistochemistry on transverse sections through the trunk regions of E9.5 and E10.5 using antibodies against CREB and P-CREB reveals that CREB staining is ubiquitous, whereas P-CREB staining is selective. (PDF 188 kb)

Supplementary Figure S2

CREB-/-S embryos have smaller somites because of a reduction in the rate of cell proliferation, rather than an increase in apoptosis. (PDF 759 kb)

Supplementary Figure S3

Wnt1 and Wnt7a expression is normal in CREB mutants. (PDF 131 kb)

Supplementary Video S4

Gene delivery into mouse embryonic somites through adenovirus infection. Lateral view of E9.75 mouse microinjected with purified adenovirus (labelled with methyl green dye) into the somites on the right side of the body. Timelapse covers a period of approximately 2 minutes. (MOV 606 kb)

Supplementary Figure S5

ACREB adenovirus does not affect expression of Pax1 (marker of developing sclerotome) in injected embryos. (PDF 70 kb)

Supplementary Figure S6

WNT1 and WNT7a induction of CRE-Luc reporter activity in 10T1/2 and C57MG reporter lines is primarily mediated by AC, rather MAPK, PKC, Ca2+ (also known to converge on CREB), or other WNT signalling components, including JNK and canonical effectors. (PDF 21 kb)

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Chen, A., Ginty, D. & Fan, CM. Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins. Nature 433, 317–322 (2005). https://doi.org/10.1038/nature03126

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