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Marvin MJ, Di Rocco G, Gardiner A, Bush SM, Lassar AB.
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In the chick, heartmesoderm is induced by signals from the anterior endoderm. Although BMP-2 is expressed in the anterior endoderm, BMP activity is necessary but not sufficient for heart formation. Previous work from our lab has suggested that one or more additional factors from anterior endoderm are required. Crescent is a Frizzled-related protein that inhibits Wnt-8c and is expressed in anterior endoderm during gastrulation. At the same stages, expression of Wnt-3a and Wnt-8c is restricted to the primitive streak and posteriorlateral plate, and is absent from the anterior region where crescent is expressed. Posteriorlateral plate mesoderm normally forms blood, but coculture of this tissue with anterior endoderm or infection with RCAS-crescent induces formation of beating heartmuscle and represses formation of blood. Dkk-1, a Wnt inhibitor of a different protein family, similarly induces heart-specific gene expression in posteriorlateral plate mesoderm. Furthermore, we have found that ectopic Wnt signals can repress heart formation from anteriormesoderm in vitro and in vivo and that forced expression of either Wnt-3a or Wnt-8c can promote development of primitive erythrocytes from the precardiac region. We conclude that inhibition of Wnt signaling promotes heart formation in the anteriorlateralmesoderm, whereas active Wnt signaling in the posteriorlateralmesoderm promotes blood development. We propose a model in which two orthogonal gradients, one of Wnt activity along the anterior-posterior axis and the other of BMP signals along the dorsal-ventral axis, intersect in the heart-forming region to induce cardiogenesis in a region of high BMP and low Wnt activity.
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Figure 2.Crescent is an efficient Wnt-8 antagonist. (A) Crescent injection into one cell of a two-cell embryo enlarged anterior structures and inhibited posterior extension in injected Xenopus embryos (bottom series of embryos). Control embryos (top) were injected with globin mRNA. LacZ mRNA was in- cluded as a lineage tracer. (B) Crescent inhibited the induction of siamois by chick Wnt-8c in Xenopus animal caps. RT–PCR analysis of siamois and ornithine decarboxylase (ODC) expres- sion in whole embryos (WE; lane 1) or animal caps from em- bryos injected with the following RNAs: globin RNA (lane 2), 1 ng of crescent RNA (lane 3), 200 pg of chick Wnt-8c RNA (lane 4), 200 pg of chick Wnt-8c and 1 ng of crescent RNA (lane 5), 10 pg of mWnt-3a (lane 6), or 10 pg of mWnt-3a and 1 ng of crescent (lane 7). All injected RNA was made up to 1.2 ng with globin RNA. The difference between the levels of siamois expression in lanes 6 and 7 was approximately threefold, when normalized to ODC levels.
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