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. 2016 Jun 1;9(6):607–620. doi: 10.1242/dmm.024661

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© 2016. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 2. — Knockdown of DS-epi1 reduces DS epimerase activity and NC-derived structures. (A) Morpholino oligonucleotides target the translation initiation sites of Dse and Dsel. (B) Endogenous DS epimerase activity is substantially decreased by Dse-MO but only a little by Dsel-MO in stage 25 embryos. (C) Epimerase activity induced by the injection of 1 ng Dse mRNA is blocked by Dse-MO, but not by control-MO and Dsel-MO. The activity of 1 ng non-targeted Dse* mRNA is not affected by Dse-MO. Results in B and C are mean±s.d. (n=3). (D) Tadpole at stage 40 injected with control-MO. (E,F) Microinjection of Dse-MO, but not Dse-5MM-MO, induces small eyes, a lack of dorsal fin structures (arrowheads) and reduced melanocyte formation (arrow). (G,H) Transversal trunk sections of stage 38 embryos following hematoxylin and eosin staining. Note the lack of a dorsal fin (arrowhead), dorsally approaching somites and hypoplastic notochord in the Dse-morphant embryo. (I-K) Ventral view of head skeletons at stage 45 in a schematic overview (I) and following Alcian Blue staining (J,K). Injection of Dse-MO, but not control-MO, causes a reduction of NC-derived cartilage structures. br, branchial segment; hy, hyoid segment; df, dorsal fin; ma, mandibular segment; no, notochord; nt, neural tube; so, somite. The proportion of examined tadpoles or explants with the indicated phenotype was as follows: D, 70/70; E, 71/114 (microcephaly), 92/114 (reduced dorsal fin), 70/114 (less melanocytes); F, 63/63; G, 4/4; H, 4/4; J, 25/25; and K, 20/20.