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Fig. 3. Ectopic Xema inhibits mesendoderm induction. (A) Xema misexpression
disrupts embryonic development. Lateral views of stage 35 embryos, anterior is
to the right. (B) Whole-mount immunohistochemistry of tailbud stage embryos
using the somite-specific antibody 12/101; lateral views, anterior is to the
right. Embryos were stained with Red Gal as a substrate prior to
immunohistochemistry. (C) RT-PCR analysis of dorsal (DMZ) and ventral (VMZ)
marginal zone explants from embryos injected with Xema RNA, harvested
immediately after dissection at early gastrula stages. (D) Whole-mount in situ
hybridization of midgastrula stage embryos, using an antisense Xenopus
brachyury probe; vegetal pole views are shown. Embryos were stained with
Red Gal as a substrate prior to in situ hybridization. The embryos shown at
the top of the figure were co-injected with lacZ RNA and
Xema RNA in two dorsal or ventral blastomeres, at the four-cell
stage, as listed; note the presence of ß-galactosidase activity (red,
arrows) in the gap in Xenopus brachyury expression (blue). The
embryos shown at the bottom of the figure were injected with lacZ RNA
in the same regions, as indicated; note the overlap (arrows) between
Xenopus brachyury expression and ß-galactosidase activity. (E)
Inhibition of Activin-mediated mesendoderm induction by Xema. RT-PCR analysis
of animal cap explants dissected at late blastula stages and cultured until
midgastrula stages. Xema RNA was injected into the animal pole region
of both blastomeres at the two-cell stage. Activin (0.5 ng/ml) was added to
stage 9 animal caps, as listed. (F) Inhibition of Fgf-mediated mesoderm
induction by Xema. bFgf (10 ng/ml) was added to stage 9 animal caps, as
listed. For all experiments in this figure, 1 ng of Xema RNA, and/or
100 pg lacZ RNA was injected, as listed.
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