XB-IMG-43879
Xenbase Image ID: 43879
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Figure 1. Xpk and Zpk Antisense Morpholino Oligos (MO) Inhibit Convergent Extension Movements(A) Whole-mount in situ hybridization of Xpk in staged embryos; early gastrula stage 10.5 (a and b, vegetal and lateral views, respectively), late gastrula stage 13 (c and d, vegetal and dorsal views, respectively), and early neurula stage 15 (e and f, posterior and dorsal views, respectively). In (a), (c), and (e), the dorsal side is at the top. In (b), the dorsal side is at the right.(B) Northern blot analysis of Xpk. The numbers at the top of each lane indicate developmental stages. Two Xpk transcripts of around 5 kb are detected in eggs to stage 32 embryos. ODC is the RNA loading quantity control.(C) Animal cap assay; Xpk expression is induced in animal cap cells in a dose-dependent manner by the overexpression of Xbra.(D) mo-Xpk injection perturbs the correct gastrulation movements and activin-induced elongation of animal cap explants. (a) Uninjected sibling embryo at the tadpole stage (left panel) and 0.5 pg activin mRNA-injected animal cap explants (right panel). (b) Unrelated mo has no effect when injected dorsally (left panel) or in the animal cap elongation assay (right panel). (c) 4mis mo-Xpk has little effect when injected dorsally (left panel) or in the animal cap elongation assay (right panel). (d) mo-Xpk 10-pmol dorsally injected embryo (left panel) and animal cap coinjected with activin and mo-Xpk (left panel).(E) mo-Xpk prevented the translation of overexpressed Xpk-GFP, but unrelated mo or 4mis mo-Xpk had no or little effects, respectively. Translation of coinjected GFP mRNA as a negative control was not prevented.(F) In the animal cap elongation assay, mo-Xpk does not interfere with mesoderm induction (Xbra expression) or expression of either Xwnt11 or Xpk itself. Histone H4 is the cDNA loading quantity control.(G) Zpk knockdown with mo-Zpk inhibits dorsal convergence. Phenotypes are observed at the10 somite stage, 17 hpf ([a and b], lateral view; [c–f], dorsal view; [g and h], anterior view) and prim-20 stage, 33 hpf (i–k). (a, c, e, g, and i), uninjected controls. (b, d, f, h, and j), mo-Zpk 0.5 pmol-injected phenotypes. These phenotypes were similar to mo-Zpk version 2 that recognizes the distinctive site of Zpk 5′-UTR (our unpublished data). (k) 4mis mo-Zpk-injected phenotype. (e,f) Whole-mount in situ hybridization of myoD (red). Image published in: Takeuchi M et al. (2003) Copyright © 2003. Image reproduced with permission of the Publisher, Elsevier B. V.
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