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Fig. 8. Rusc1 inhibits Hh signaling during Xenopus eye development. (A) Whole embryo morphology of uninjected embryos and those injected with R1-MO,
R1-5mis or R2-MO. Morpholinos (20 ng) were injected into both dorsal blastomeres at the 4-cell stage. (B) Overexpression of myc-xRusc1 rescued the
phenotypes induced by unilateral injection of R1-MO. (Left) Summary of embryos with eye defects. (Right) Images of representative embryos. A 50% or greater
reduction in eye size is considered ‘severe’; a reduction of less than 50% is considered ‘mild’. (C) RT-PCR showing the expression of gli1, ptc1, ptc2 and hhip in
animal caps. Chordin (Chd, 25 pg) was injected into the animal pole of control and R1-MO (40 ng) injected embryos at the 1-cell stage. Animal caps were
dissected at the late blastula stage and harvested at stage 22. Data are shown as mean±s.d. *P<0.05, **P<0.01. (D) In situ hybridization showing that unilateral
injection of R1-MO (20 ng) enhances the expression of gli1, and reduces the expression of pax6, rax and six3. The expression of shh was not altered by R1-MO
injection. Embryos were analyzed at stage 20. (E) In situ hybridization showing that unilateral injection of R1-MO enhances the expression of gli1 in the head
region and reduces the expression of pax6, rax and six3 at stage 33. Arrowheads point to eyes on the injected side. (F) Morphology of uninjected embryos
and those unilaterally injected with R1-MO alone or R1-MO together with Gli1 morpholino (Gli1 MO). Insets show further examples of the illustrated phenotype.
Arrows (D,F) point to the developing eyes. |
