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Fig. 1. Misexpression of XSRF suppresses mesodermal cell fates. (A-C)
Spatial expression of XSRF at the four-cell, blastula and early gastrula
stages. Lateral views are shown. (D) XSRF expression in dissected
gastrula embryos. Animal cap (AC), vegetal pole (VP), dorsal marginal zone
(DMZ) and ventral marginal zone (VMZ) explants were dissected from stage 10.5
gastrula embryos and subjected to RT-PCR analysis. -RT, control RT-PCR in the
absence of reverse transcriptase. The following controls were included as
markers for the dissected tissues: Chordin for dorsal mesoderm, Xbra for
pan-mesoderm, XMsx1 for animal cap and ventral mesoderm and XSox17ß for
endoderm. ODC, ornithine decarboxylase loading control. (E-G)
Phenotypic effects of overexpression of wild-type (wt) XSRF RNA. Four-cell
stage embryos were injected in the dorsal (F) or ventral (G) marginal regions
with wt XSRF mRNA (2 ng), or not injected as a control (E), and cultured until
sibling embryos reached stage 30. Lateral views are shown and anterior is to
the left. (H-P) Ectopic wt XSRF interferes with the expression of
mesodermal genes. Four-cell stage embryos were injected in the dorsal or
ventral marginal regions with wt XSRF mRNA (2 ng) and, along with uninjected
controls, cultured to stage 10.25-10.5 (H-M,P) and 28 (N,O) and then analyzed
by whole-mount in situ hybridization using antisense Xenopus Wnt8 (H,I),
brachyury (J,K), chordin (L,M) and myoD (N,O) probes or RT-PCR (P). Arrowheads
in I,K,M,O indicate the absence or reduction of expression of genes. (H,I)
Ventro-vegetal views are shown. (J-M) Vegetal views are shown with dorsal side
at top. (N,O) Lateral views are shown with anterior side to the left. (P)
Ventral marginal zone explants were dissected at stage 10.25 and subjected to
RT-PCR. VMZ Co, uninjected ventral marginal zone tissue.
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