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Fig. 3. Misexpression of DD2 in the limb mesenchyme by injecting plasmid solutions
in the lateral plate mesoderm. Diagrams show the experimental design of
electroporation. (A) We misexpressed the EGFP gene to monitor the expression
of transgenes in the mesenchyme. (B) DD2 misexpression at stages 13-15 induced
repression of Fgf10 expression at stage 20 (red arrows). (C)
Wnt5a was also repressed (red arrow), with deformity of the AER.
(D-F) At stage 19, expression of Fgf8, En-1 and Msx2 became
unclear and distorted. (G) At stage 24, a broad and irregular AER was formed
as judged by the Fgf8-positive domain, in which an
Fgf8-negative line was observed. (H-K) At stage 20, Wnt7a
and Lmx1b were expressed dorsally, and Bmp7 and
Msx2 ventrally, without disturbance of the DV axis formation.
Nonetheless, thickening of the ectoderm was observed near the margin of the
expression of these markers (red arrowheads). Approximately 35% of the
DD2-misexpressed limb buds displayed this phenotype in the AER
(n=156).
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