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. 2021 Mar 23;11:6607. doi: 10.1038/s41598-021-86153-x

Figure 2.

Figure 2

Dorsal fry-depletion affects early expression of organizer genes and causes gastrulation defects. (a–l’’) In situ hybridization of Xenopus embryos at the indicated stages. (a–a’’) Expression of otx2 in the presumptive prechordal mesoderm. (a) Uninjected embryo (N = 2; n = 33). (a’) St-MO injected embryo (N = 2; n = 33; 6% with reduced expression domain) (a’’) fry-MO injected embryo (N = 2; n = 26; 85% with reduced expression domain). (b,c’’) Expression of gsc in the presumptive prechordal mesoderm. (b) Uninjected St.10 embryo (N = 3; n = 98). (b’) fry-MO injected St.10 embryo (N = 2; n = 34; 6% with reduced expression domain) (b’’) fry-MO injected St.10 embryo (N = 3; n = 74; 78% with reduced expression domain). Dorsal is oriented to the top. (c) Uninjected St.13 embryo (N = 3; n = 60). (c’) St-MO injected St.13 embryo (N = 2; n = 30; 10% with abnormally positioned expression domain). (c’’) fry-MO injected St.13 embryo (N = 3; n = 62; 87% and 96% with reduced and abnormally positioned expression domain, respectively).The distance between the blastopore and the prechordal mesoderm expressing gsc (dashed line) is reduced in the morphants. Dorsal views, anterior is oriented to the top. (d,e’’) Expression of chrd in the presumptive axial mesoderm. (d) Uninjected St. 10 embryo (N = 5; n = 95). (d’) St-MO injected St. 10 embryo (N = 2; n = 34; 6% with reduced expression domain) (d’’) fry-MO injected St. 10 embryo (N = 5; n = 113; 97% with reduced expression domain). Dorsal is oriented to the top. (e) Uninjected St.13 embryo (N = 4; n = 128). (e’) St-MO injected St.13 embryo (N = 2; n = 30, 10% with abnormally positioned expression domain) (e’’) fry-MO injected St.13 embryo (N = 4; n = 115, 92% and 95% with reduced and abnormally positioned expression domain, respectively). Dorsal views, anterior is oriented to the top. (f–h’’) Expression of pan-mesodermal marker brachyury (xbra). (f) Uninjected St. 10.5 embryo (N = 3; n = 47). (f’) St-MO injected St. 10.5 embryo (N = 2; n = 31). (f’’) fry-MO injected St. 10.5 embryo (N = 3; n = 53). (g) Uninjected St. 13 embryo (N = 2; n = 44). (g’) St-MO injected St. 13 embryo (N = 2; n = 28; 7% with abnormally positioned expression domain) (g’’) fry-MO injected St. 13 embryo (N = 2; n = 52; 98% with abnormally positioned expression domain). (h) Uninjected St. 15 embryo (N = 3; n = 48). (h’) St-MO injected St. 15 embryo (N = 2; n = 33). (h’’) fry-MO injected St. 15 embryo (N = 3; n = 45). Dorsal views, anterior is oriented to the top. (i–l’’) Expression of not in the chordamesoderm and anterior neuroectoderm. (i) Uninjected St. 10.5 embryo (N = 2; n = 52). (i’) St-MO injected St. 10.5 embryo (N = 2; n = 31). (i’’) fry-MO injected St. 10.5 embryo (N = 2; n = 47). (j) Uninjected St. 13 embryo (N = 3; n = 39). Dorsal is oriented to the top. (j’) St-MO injected St. 13 embryo (N = 2; n = 27; 4% abnormally positioned expression domain). (j’’) fry-MO injected St. 13 embryo (N = 3; n = 43; 100% with abnormally positioned expression domain). (k) Uninjected St. 13.5 embryo (N = 2; n = 29). (k’) St-MO injected St. 13.5 embryo (N = 2; n = 24). (k’’) fry-MO injected St. 13.5 embryo (N = 2; n = 32, 63% with abnormally positioned expression domain). (l) Uninjected St. 15 embryo (N = 3; n = 46). (l’) St-MO injected St. 15 embryo (N = 2; n = 28). (l’’) fry-MO injected St. 15 embryo (N = 3; n = 59). Dorsal views, anterior is oriented to the top. Note: not expression in the epiphysis (arrowheads) is initiated in fry-depleted embryos at the same time as controls, indicating the morphants are not developmentally delayed. Dashed lines indicate dorsal mesoderm elongation. The stage of injected embryos was established based on the stage of uninjected embryos from the same clutch. Representative embryos are shown. Embryos were injected into both dorsal blastomeres at the 4-cell stage with 15 ng of fry-MO or St-MO.