Figure 2: Afp18G disturbs zebrafish early development. | Nature Communications

Figure 2: Afp18G disturbs zebrafish early development.

From: Tyrosine glycosylation of Rho by Yersinia toxin impairs blastomere cell behaviour in zebrafish embryos

Figure 2

(a) Live images of non-injected, GFP, Afp18G NxN mRNA (each 100 pg per embryo), or different amounts (0.1–100 pg per embryo) of Afp18G mRNA-injected embryos at 1,000-cell stage (3 h.p.f.). Embryos oriented animal to the top. Scale bar, 500 μm. (b) Quantification of the blastoderm phenotype of control and Afp18G-injected embryos shown in a at 1,000-cell (non-injected, n=41; GFP mRNA, n=37; Afp18G NxN mRNA, n=52; 0.1 pg Afp18G, n=35; 0.5 pg Afp18G, n=45; 1 pg Afp18G, n=60; 10 pg Afp18G, n=62; 100 pg Afp18G, n=60). Only values for 100 pg per embryo injected GFP and Afp18G NxN mRNA are shown. Developing live embryos were classified into categories ‘normal’ (WT like), ‘disrupted blastoderm’ (cellular structure of blastoderm abnormal) or ‘disintegrated embryos’ (blastoderm and (or) yolk cell lysed). The distribution of phenotypes was analysed for significant differences using Fisher exact probability test, revealing significant differences (*P values <0.0001) between Afp18G NxN control and Afp18G samples. (c) Optical image planes of live blastoderm regions at 1,000-cell (3 h.p.f.) of non-injected, GFP, Afp18G NxN or Afp18G mRNA-injected embryos (pg per embryo) oriented animal to the top. Control embryos show normal development of blastomeres, while the blastoderm progressively loses cellular integrity with increasing amounts of Afp18G mRNA injected. Arrowheads mark the disrupted regions without visible blastomere boundaries, abnormal sized and irregular shaped cells. Asterisks mark detached blastomeres and disintegrated blastoderm. Scale bar, 100 μm. (d) Live images of control, full-length Afp18 and full-length Afp18 NxN mRNA (each 50 pg per embryo) injected zebrafish embryos. Afp18-injected embryos disintegrate at 1,000-cell stage (3 h.p.f., upper row, embryos are oriented animal to the top) and degrade early in gastrulation (lower row 40% epiboly, 5 h.p.f.). Scale bar, 500 μm. For five different concentrations each of Afp18 (non-injected n=80; 2.5 pg n=34; 5 pg n=50; 12.5 pg n=65; 25 pg n=85; 50 pg n=38) and Afp18 NxN (2.5 pg n=43; 5 pg n=58; 12.5 pg n=50; 25 pg n=76; 50 pg n=23) mRNA injections, the bar graph shows percentage of embryos, which at 5 h.p.f. develop like WT controls or degrade.

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