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Fig. 7. Phenotypic characterization of ezrin2 morphant embryos. (A) Images of wild-type (wt) and ezrin2-MO1 injected embryos at 50% epiboly (5 hpf) and 60% epiboly (6 hpf). Dorsal side is to the right and the animal pole is up. (B) Face-on views of blastodermal cells within the animal pole of 50% epiboly (5 hpf) wild-type (wt) and ezrin2 morphant (MO1) embryos as DIC images and stained for F-actin with phalloidin. Bar, 20 µm. (C) Images of wild-type (wt) and ezrin2-MO2 injected embryos at 80% epiboly (8 hpf) and bud stage (10 hpf). Dorsal side is to the right and the animal pole is up. (D) In situ hybridization of wild-type (wt) and ezrin2-MO2 injected embryos at the bud stage (10 hpf) with markers delineating convergent extension of the forming body axis. Upon morpholino injection, embryos developed a shortened notochord and a broadened neural plate. Animal views (upper panel) and dorsal views (lower panel), anterior to the left. Markers: notochord, notail (ntl); anterior edge of neural plate, distal-less homeobox 3 (dlx3); prechordal plate, hatching gland gene-1 (hgg1). (E) Analysis of cellular rearrangements within the axial germ ring (shield) of wild-type and ezrin2 morphant embryos (MO2) starting at shield stage (6 hpf) by confocal time-lapse microscopy. Images correspond to timepoints 0 minutes and 60 minutes of Movies 1 and 2 in supplementary material. One exemplary mesendodermal progenitor cell was labeled in red to mark its position at 0 and 60 minutes of the time-lapse movie. Note that during this time interval the labeled cell has moved further away from the germ ring margin in the wild-type embryo compared with the morphant embryo, suggesting that mesendodermal cell migration is reduced within the shield of ezrin2 morphant embryos. Lateral view, dorsal to the right. Bar, 40 µm.
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