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Fig. 3. Depletion of kermit 2/XGIPC specifically inhibits Xenopus eye
development. (A-D) Depletion of kermit 2 does not affect the
expression of dorsal mesoderm markers chordin and goosecoid.
Embryos are viewed from the vegetal side and dorsal is towards the top.
Control morpholino or kermit 2 morpholino (40 ng) was injected into two dorsal
animal blastomeres of four-cell embryos. Embryos were fixed at the gastrula
stage (stage 10) and whole-mount in situ hybridization was performed for
chordin (A,B) and goosecoid (C,D). (E-L) Depletion of
kermit 2 specifically reduces marker gene expression within the presumptive
eye field in stage 20 embryos. Embryos are viewed from the anterior side with
dorsal towards the top. Control or kermit 2 morpholino was injected into one
dorsal animal blastomere of four-cell/eight-cell embryos with 500 pg of mRNA
for nuclear ß-galactosidase. Embryos were fixed at stage 20 and
ß-galactosidase activity was measured in situ (red) followed by
whole-mount in situ hybridization for Bf1 (E,F), Otx2 (G,H),
Pax6 (I,J) and Xrx (K,L). Bf1 expression was not
affected by depletion of kermit 2 (F). Expression of Otx2 (H) and
Pax6 (J) were reduced only within the presumptive eyeforming region
(red arrowheads) in embryos injected with kermit 2 morpholino. The expression
of the eye marker Xrx was also inhibited (L red arrowhead).
(M-P) Depletion of kermit 2 does not reduce Xrx (N) or
Pax6 (P) expression in early neurula stage embryos (stage 14).
(Q-S) Kermit 2 mRNA restored Xrx expression in kermit
2-depleted embryos. Red arrowhead in R indicates strongly reduced Xrx
expression within presumptive eye domain in kermit 2-depleted embryo; red
arrow in S indicates recovered Xrx expression in embryo co-injected
with kermit 2 morpholino and kermit 2 mRNA.
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