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Figure 2. In situ hybridization analysis for putative targets of Pitx3 involved in eye development. Visual comparisons of gene expression patterns between right-side injected control-morpholino (Cmo) or Pitx3-morpholino (Pmo) embryos and their untreated contralateral control. A–E: Pitx3 expression patterns are presented for comparison (adapted from Khosrowshahian et al., 2005; Smoczer et al., In Press). A: demonstrates faint but detectable signal throughout the ectoderm and in agreement with reverse transcriptase-polymerase chain reaction (RT-PCR) results. B: Expression is detectable throughout neural ridge, while at stage 22, the gene is expressed in a cleared specimen where an arrow indicates presomitic mesoderm. By stage 27 (D), Pitx3 is detectable throughout much of the head ectoderm, as well as in branchial arches and somites. This pattern restricts later to somites, otic vesicle, lens, and brain (D). F–G′: Vent2 expression is reduced in the developing eye field at stage 19 for the Pitx3-morpholino (Pmo) injected side (A′ white arrow) and at stage 27 (B′ black arrow), when compared with control-morpholino (Cmo) injected embryos (A,B). H–I′: Pax6 shows reduced expression in eye field on Pmo side of embryos at stage 19 (C′ black arrow) and 27 (D′ white arrow). J–J′: Crybb1 shows drastic loss of expression in the eye vesicle on the Pmo-treated side of stage 27 embryo (E′) and no difference caused by Cmo treatment (E). Dotted line represents the midline of the embryo, separating injected right side from contralateral left side control.Download figure to PowerPoint |
