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Figure 1. Loss of Cd63 in the neural ectoderm caused defects in eye morphogenesis: A Expression pattern of cd63 during tailbud stages. Enrichment of transcripts in the TNC of st. 25 (white arrowheads). At st. 30, cd63 was additionally detected in eyes and cement gland. Transversal sections highlighted signals in the eyes, head mesenchyme, brain, premigratory TNC, dorsal fin mesenchyme, medially and laterally migrating TNC cells (black arrowheads). cd63 sense control specimen of st. 29 did not show any signal. B Knockdown of cd63 in one dorsal animal blastomere resulted in impaired eye development, which was rescued by co-injection of cd63 mRNA. Sections illustrated normal eye structures in control and rescue specimen, whereas morphant embryos depicted disturbed optic cup formation on the injected side. Quantification of eye phenotypes illustrates significance of the effect. C Unilateral loss of cd63 in the eye field lineage revealed altered expression pattern of otx2 in neurula and later tailbud stages. At neurulation, otx2 showed no reduction in intensity but indicated alterations of the targeted eye vesicle (black arrowhead). Sections revealed defects in medial forebrain closure and lateral optic vesicle formation. In tailbud stages, loss of cd63 resulted in partial reduction of eye tissues (black arrowhead) compared to matching controls. Both effects were reduced by co-injection of cd63 mRNA (white arrowheads). D cd63 TBMO injection did not block TNC migration or dct expression in eyes or melanophores of unilateral morphants. Sections revealed massively impaired formation of eye tissue (black arrowhead) compared to WT eyes of controls. Asterisks mark the injected side. a, anterior; co, control; d, dorsal; p, posterior; TBMO, translation blocking Morpholino oligomer; TNC, trunk neural crest; st., stage; v, ventral; wt, wild type. |
