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Fig. 1

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ZDB-IMAGE-201113-7
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Figures for Bosze et al., 2020
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Fig. 1

A cell group in the axial mesoderm regulates notochord morphogenesis in zebrafish. a Characterization of the prechordal plate (ppl) by whole-mount double in situ hybridization (ISH) of wild-type (WT) zebrafish embryos with the indicated markers. Scale bar: 100 µm. b Mapping of pcdh18a mRNA expression (red) relative to Tg(gsc:GFP) expression labelled by anti-GFP antibody (green). Notably, gsc:GFP labels the prechordal plate (ppl) and the trailing notochord (NC). Scale bar: 100 µm. c Inhibition of nodal signalling by SB505124 treatment (30 µM) from 4 to 8.5 h post-fertilization (hpf). Scale bar: 100 µm. d Confocal image of live zebrafish embryo at 5 hpf showing subcellular localization of Pcdh18a-GFP. Glycosylphosphatidylinositol-anchored mCherry (memCherry) marks cell membranes. Arrows indicate punctae of Pcdh18a localization at the membrane (open arrows) and intracellularly (closed arrows). Scale bar: 10 µm. e WT embryos or Tg(gsc:GFP) embryos were injected with Pcdh18a MO (0.5 mM). Morpholino-based knockdown of Pcdh18a leads to a wider and shorter notochord marked by ntl expression at 9 hpf (arrows). See Supplementary Fig. S1h for control experiments. Analysis of the shape of the notochord in a cross section of gsc:GFP transgenic embryos that were injected with the indicated constructs at 10 hpf. At 11 hpf, the body length was significantly shorter in the Pcdh18a-deficient embryos, as shown in an ISH-based analysis of notochord hgg/ntl (n = 20/32, arrows). Confocal microscopy-based analysis of cell shapes in the notochord of embryos that were microinjected with the indicated constructs at 12 hpf (cells with exemplary morphology were surrounded with a yellow circle). Scale bar: 100 µm. fpcdh18a MO-injected embryos show a wider axial mesoderm compared to Ctrl MO injected embryos (four embryos each). g Circularity of NC cells was measured in total of 1000 cells in five different embryos each. A circle has a circularity of 1.0, while noncircular shapes have a lower value of circularity. The error bars represent the SEM and significance as indicated (***p value < 0.001; unpaired Student’s t test)

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This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Histochem. Cell Biol.