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Fig. 2. VP=Dach1 was misexpressed in the ectoderm by pouring the plasmid solution
over the ectoderm. Diagrams at the top show the experimental design of
electroporation. (A) A dorsal view of an electroporated limb bud on the
right-hand side (Exp.) compared with a normal limb bud on the left. Expression
of Fgf8 is obscure. (B) In a lateral view, Fgf8 expression
is faint and blurred without making a clear boundary. (C) At stage 24, a
severe depression was evident in the dorsal end of the limb bud. (D) Strong
EGFP signals, which were derived from co-electroporated pCAGGS-EGFP, were
observed in the ectoderm. (E) In such limb buds, Fgf8 expression was
lost in the central part and deformed in the anterior (red arrowheads),
whereas Fgf8 was normally expressed in the posterior (blue
arrowhead). (F) Expression of Bmp7 was also lost in the central part
(red arrowhead) and in the anterior mesenchyme, but was normal in the
posterior area (blue arrowhead). (G) Mesenchymal expression of Fgf10
was repressed with inhibition of the limb outgrowth (red arrowheads). (H)
Shh was expressed normally posteriorly (blue arrowhead). A depression
similar to C was evident in the distal end (red arrowhead). (I) At stage 37,
distal truncation was obtained. In the wing, where misexpression of VP=Dach1
was weak, truncation of digit II was evident. In the leg, where misexpression
was extensive, distal autopod structures were completely missing (red
arrowhead). (J) Alcian Blue staining of these limbs revealed a short digit II
in the wing (red arrow) and the complete loss of digits in the leg (red
arrowhead). Approximately 60% of the misexpressed limbs showed these
phenotypes (n=242).
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