The novel Cer-like protein Caronte mediates the establishment of embryonic left–right asymmetry

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In the chick embryo, left–right asymmetric patterns of gene expression in the lateral plate mesoderm are initiated by signals located in and around Hensen's node. Here we show that Caronte (Car), a secreted protein encoded by a member of the Cerberus/Dan gene family, mediates the Sonic hedgehog (Shh)-dependent induction of left-specific genes in the lateral plate mesoderm. Car is induced by Shh and repressed by fibroblast growth factor-8 (FGF-8). Car activates the expression of Nodal by antagonizing a repressive activity of bone morphogenic proteins (BMPs). Our results define a complex network of antagonistic molecular interactions between Activin, FGF-8, Lefty-1, Nodal, BMPs and Car that cooperate to control left–right asymmetry in the chick embryo.

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Figure 1: Expression of Car, Nodal and Lefty-1 during chick gastrulation.
Figure 2: Car acts downstream of Shh and upstream of Nodal and directs heart looping in the chick embryo.
Figure 3: Antagonism of BMPs by Car regulates Nodal expression.
Figure 4: Interaction of Car with BMPs: biochemical evidence and structural model.
Figure 5: FGF-8 and Activin regulate Car expression.
Figure 6: Interaction between Lefty-1 and Car during the establishment of left–right asymmetry.
Figure 7: cNKX3.2 is a novel target of the left–right signalling pathway.
Figure 8: Role of Car in the genetic cascade that determines left–right development.


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We thank J. Magallón for technical help; T. Brand, R. Evans, H. Hamada, C. Kintner, G. Rosenfeld, C. Stern, C. Tabin and members of P. E. Wright's group for discussions and for sharing unpublished observations; W. Vale and S. Choe for reagents; J. P. Fandl (for providing human Cerberus) and X. Wang (for technical help), both at Regeneron; J. Hurlé for scanning images; C. Parada for suggesting the name Caronte; and L. Hooks for help in preparing the manuscript. J.C. was supported by a Hoffmann Foundation Fellowship; J.P. was supported by a long-term EMBO Fellowship. This work was supported by grants from the G. Harold and Leila Y. Mathers Charitable Foundation and the NIH to J.C.I.B., who is a Pew Scholar.

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Correspondence to Juan Carlos Izpisúa Belmonte.

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