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The opacity in the PD0332991 and optic vesicle of yap morphants suggests a distinct tendency towards cell death ( Fig. 2IID and E). The TUNEL assay revealed that yap-MO causes a marked increase in cell death in the brain ( Fig. 3I and J).
Craniofacial cartilage defects in yap morphants
Embryos injected with yap-MO develop craniofacial abnormalities that are apparent at 96 hpf ( Fig. 4). Alcian blue staining of cartilaginous structures showed that the shortened Meckel’s cartilage and the ceratohyal arch resided more posteriorly in yap morphants than in embryos injected with con-MO, while other cartilaginous structures, including the ceratobranchial arches, were weakly stained or not visible at all ( Fig. 4A–B′). In addition, in the most severely affected embryos, no signal for chondrogenesis was detected (data not shown).
Fig. 4.
Abnormal branchial arch development in yap morphants. Embryos injected with con-MO (5 ng) (A,A′) or yap-MO (5 ng) (B,B′) at 96 hpf, stained with Alcian blue to reveal craniofacial cartilages. Ventral (A,B) and lateral (A′,B′) views show that the shortened Meckel’s cartilage and the ceratohyal arch resided more posteriorly in yap morphants than in embryos injected with con-MO, while other cartilaginous structures, including the ceratobranchial arches, were weakly stained or not visible at all. (C,D′) In yap morphants at 30hpf, expression of crestin was markedly reduced in all recognizable arch-associated regions, which are indicated by arrows. In addition, a population of cranial NCCs was distributed abnormally in the dorsal anterior periocular mesenchyme, with few NCCs migrating to the eye field. Abbreviations: cb1–5, ceratobranchial arches 1–5; ch, ceratohyal arch; mk, Meckel’s cartilage.