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. 2019 Oct 30;9:15645. doi: 10.1038/s41598-019-52218-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The caveolar core components caveolin1, caveolin3 and cavin1 are necessary for convergent extension movements. Antisense morpholino oligonucleotides directed against caveolin1 (cav1Mo), caveolin3 (cav3Mo) and cavin1 (cavin1Mo) were injected in the marginal zone of both dorsal blastomeres of 4-cell stage embryos. Embryos were fixed at stage 14 and analyzed for localization of the axial mesodermal marker gene chordin. (a) In control embryos, chordin is expressed in a long and narrow strip along the dorsal midline. Depletion of caveolar proteins resulted in mislocalization of chordin expression. In severe cases, chordin expression remained at the dorsal part of the unclosed blastopore. The phenotype was classified as mild when the chordin expression was bent or short and broad. (b) Quantification of the phenotypes in the morphants resulting from the injection of 4 pmol cav1Mo, 4 pmol cav3Mo and 0.4 pmol cavin1Mo, respectively. (c) Caveolin1 and cavin1 patially restore chordin expression in cav1 morphants. 4 pmol cav1Mo were coinjected with 500 pg caveolin1 and cavin1 mRNA, respectively. (d) Epistasis experiments with Wnt5a morpholino (5aMo, 0.4 pmol), caveolin1 morpholino (cav1Mo, 0.25 pmol) and caveolin1 mRNA (500 pg) revealed additive effects of the morpholinos. Furthermore, caveolin1 mRNA partially restored gastrulation in Wnt5a morphants. P values according to a χ² test, n: number of embryos. (e) Realtime PCR of animal cap explants revealed that the expression of the Wnt5a target gene PAPC depends on caveolin1 expression. Shown are the mean values, standard errors and p values according to Student’s t-test of four independent experiments.