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Fig. 4. Ectopic expression of Bmps in Smad5^m1/m1 amnion. (A-F) Ectopic expression of Bmp2 and Bmp4 in mutant amnion at E8.5. Compared with the wild-type littermate (A,B), the expression of Bmp2 is highly elevated in the aggregate of cells (KO, arrow) and the flanking mutant amnion, but not in more remote squamous amnion (arrowhead) (C,D), or in surface ectoderm. Smad5 ^m1/m1 embryos express much higher levels of Bmp4 throughout the amnion (F), than do controls (E). Bmp4 expression levels appear weaker in mutant surface ectoderm (E,F). Surface ectoderm and amnion ectoderm are often poorly delineated in Smad5 ^m1/m1 embryos when the aggregate of cells on the amnion is localized anteriorly (dashed circle; C). (G-L) Enhanced expression of Bmp4, but not of Bmp2, in mutant amnion at E7.0. Bmp4 is expressed in wild-type amnion at EB and NP stages, respectively (G,I), but its expression is elevated in the amnion of mutant littermates (H,J). The Bmp2 expression is not yet significantly different in the amnion of wild-type (K) and mutant (L) EB littermates. Short arrows point towards the mesodermal component of the amnion, and are positioned in the exocoelom. Anterior is to the left and posterior to the right. AmMe, amnion mesoderm; He, heart; Nt, neural tube; Ps, primitive streak; Se, surface ectoderm; Ys, yolk sac. Scale bars: 100 µm in B-H,J,L; 50 µm in A,I,K.

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