Figure 1

PMP treatments cause stage dependent effects on eye DV polarity. (A) Lateral views of heads of st. 33 embryos treated with DMSO (mock) or 300 to 600 μM PMP from the indicated stages and hybridized with probes for Pax2, Vax1b, Vax2 or Tbx3. In mock-treated embryos, wild-type expression patterns of these genes are detectable: Pax2 and Vax1b staining is restricted to the OS, Vax2-positive region covers the OS and VR, Tbx3 expression identifies the DR. PMP treatments affect gene expression domains to various degrees depending on the stage of delivery (st. 8 to 10, blastula-early gastrula; st. 12.5 to 14, late gastrula-early neurula; st. 19 to 22, late neurula-early optic vesicle; st. 26, mid optic vesicle). For Pax2 and Vax1b, embryos were grouped into 0 to 3 scores (numbers at the bottom right corner of each image) as explained in the ‘Results’ section and representative eyes for each score group are shown. See text for details. The broken yellow circles highlight the eye region. Scale bar, 200 μm. (B) Quantification of the percentages of embryos stained for Pax2 or Vax1b with 0 to 3 scores in each treatment condition. Embryos stained for Vax2 or Tbx3 were grouped according to the DV extent of Vax2/Tbx3 expression domain (more or less than 90% of the eye for Vax2; more or less than 10% of the eye for Tbx3). The percentages of embryos with strong eye reductions are also indicated (S, small eyes). The number of experiments performed for each probe and treatment condition is indicated on top of the corresponding histogram bar. At least 20 eyes/10 embryos were analysed for each experiment. (C) Histological sections of eyes of st. 33 embryos treated as in (A) and (B), and hybridized with the indicated probes, confirming stage dependent alterations in the expression domains of Pax2, Vax1b, Vax2 and Tbx3 as detected in whole mount views. Scale bar, 100 μm.