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Fig. 6. The Wnt/ β-catenin signaling pathway negatively regulates
specification and differentiation of cells from the SpM and CPM in chick
embryos. (A) Schematic representation of the experimental procedure
showing the electroporation of the construct into a St. 3c embryo, which was
analyzed 16 hours later, at St. 8. (B-D) Fluorescent ISH of the St. 8
electroporated embryos for the markers indicated. As a result of Wnt3a
activity, the expression levels of Tbx5 (B), Nkx2.5(C) and
Gata4 (D) transcripts were reduced on the electroporated side
(arrows; n=14). (E) Representation of the experimental
procedure showing the electroporation of the pCIG-Wnt3a-IRES-GFP
construct into a St. 8 embryo, and its analysis at St. 12 (note a reverse
cardiac looping). (F,G) St. 12 embryos stained for Isl1 (F) and
Nkx2.5 (G) following electroporation with pCAGG-GFP
(arrows). The normal expression of both proteins was not affected
(arrowheads). (F',G') St. 12 embryos, electroporated
with pCIG-Wnt3a-IRES-GFP marked by the GFP expression in the surface
ectoderm. Both Isl1 (F') and Nkx2.5 (G') levels in the SpM of the
AHF (arrowheads) were reduced on the electroporated side (n=17).
(H-I') Whole-mount ISH for Tbx1 and capsulin in St. 13
embryos electroporated with either pCAGG-GFP or
pCIG-Wnt3a-IRES-GFP. Both Tbx1 (H') and capsulin
(I') levels in the BA were reduced (arrows) on the
Wnt3a-electroporated side (n=9). nt, neural tube; hrt,
heart; ph, pharynx.
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