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Figure 2. FoxA4MO disrupted DML formation.
(A–H) Stage 15 embryos left uninjected (A) or injected with 0.5 ng of foxA4FL mRNA (E), 40 ng of CtrlMO (B–D) or of FoxA4MO (F–H) before the first cleavage. (I–T) Tailbud stage embryos injected into two dorsal cells at the 4-cell stage with 20 ng of CtrlMO or of FoxA4MO. The descendants of these cells give rise to the DML and the cephalic region. Expression of chd (A, B, E, F, I, J, M, N, Q, R), shh (C, G, K, L, O, P, S, T), hairy2 (D, H). The inset in K shows a magnified view of the area depicted by the rectangle; fp, floor plate; no, notochord; h, hypochord. Embyos injected with foxA4FL mRNA showed a thicker notochord than control siblings, as revealed by chd expression (87%, n = 15) (E). The percentage of embryos injected with FoxA4MO showing the indicated changes in the corresponding markers is indicated between parentheses, as follows. At neural plate stage, the expression of chd (90%, n = 20) (F) and shh (54%, n = 22) (G) was patchy, and hairy2 expression was reduced in the FP domain (60%, n = 23) (H). At stage 26, the chd domain was patchy and/or thinner (80%, n = 21) (M, N, Q, R). Arrowheads point to chd + cells in dorsal and ventral positions with respect to a notochordal gap. Shh was also patchy (arrowheads, O, P) or completely abolished (S,T) (86%, n = 22). In overall, tailbuds injected with FoxA4MO showed reduced heads (89%, n = 51) (M, Q, O, S). (A–H, J, L, N, P, R, T) dorsal views; (I, K, M, O, Q, S) lateral views.
doi:10.1371/journal.pone.0110559.g002 |
