Smad2 role in mesoderm formation, left–right patterning and craniofacial development

Abstract

Signalling by the transforming growth factor-β (TGF-β) superfamily of proteins depends on the phosphorylation and activation of SMAD proteins by heteromeric complexes of ligand-specific type I and type II receptors with serine/threonine-kinase activity1. The vertebrate SMAD family includes at least nine members, of which Smad2 has been shown to mediate signalling by activin and TGF-β2,3,4,5. In Xenopus, Smad2 can induce dorsal mesoderm, mimicking Vg-1, activin and nodal2,4. Here we investigate the function of Smad2 in mammalian development by generating two independent Smad2 mutant alleles in mice by gene targeting. We show that homozygous mutant embryos fail to form an organized egg cylinder and lack mesoderm, like mutant mice lacking nodal6,7 or ActRIB, the gene encoding the activin type-I receptor8. About 20 per cent of Smad2 heterozygous embryos have severe gastrulation defects and lack mandibles or eyes, indicating that the gene dosage of Smad2 is critical for signalling. Mice trans-heterozygous for both Smad2 and nodal mutations display a range of phenotypes, including gastrulation defects, complex craniofacial abnormalities such as cyclopia, and defects in left–right patterning, indicating that Smad2 may mediate nodal signalling in these developmental processes. Our results show that Smad2 function is essential for early development and for several patterning processes in mice.

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Figure 1: The Smad2mh1 mutation results in early embryonic lethality and other defects.
Figure 2: The early developmental defects in Smad2mh2-lacZ mutant embryos.
Figure 3: Defects in egg-cylinder organization and primitive streak formation in Smad2mh1 mutant embryos.
Figure 4: Gastrulation, craniofacial and laterality defects in Smad2mh1/+, nodallacZ/+ trans-heterozygotes.

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Acknowledgements

We thank E. Robertson for nodallacZ mice; P. Ten Dijke for Smad2 antibody; A. Smith for pGT1.8Iresβgeo; P. Oh for help with whole-mount in situ hybridization; H. Lei, M. Okano and L. Yu for technical assistance; and Z. Gu and T. Gridley for comments on the manuscript. This work was funded by Bristol Myers-Squibb (E.L.) and a postdoctoral fellowship from the Japan Society for the Promotion of Science (M.N.).

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Correspondence to En Li.

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Nomura, M., Li, E. Smad2 role in mesoderm formation, left–right patterning and craniofacial development. Nature 393, 786–790 (1998). https://doi.org/10.1038/31693

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