| |
Figure 6. FoxA4 overexpression produced complementary effects on Xanf1 and otx2 and disfavoured anterior development.
(A, B, F, G, K, L) Anterior views of early neural plate stage embryos analysed by WMISH with the following markers: Xanf1 and en2 (A, F, K), otx2 and krox20 (B, G, L). Embryos were injected with 0.25 ng (F, K) or 1 ng (G, L) of foxA4FL mRNA before the first cleavage, or were left uninjected (A, B). FoxA4 overexpression affected Xanf1 expression in 83% of the injected embryos (n = 23) as follows: it was reduced in the ANR but not in the SHA (48%, n = 23) (F), or it was reduced in the whole domain (35%, n = 23) (K). The expression hole of the otx2 domain was filled with otx2 transcripts (42%, n = 19) (G); the otx2 domain was reduced, but the caudal diencephalic/mesencephalic stripes were expanded (89%, n = 19) (L). (C–E, H–J, M–O) Anterior views of tailbuds (stage 27/28) injected with 0.25 ng (H, I, M, N) or 0.5 ng (J, O) of foxA4FL mRNA before the first cleavage (I, J, M–O) or into one cell at the 2-cell stage (H), or left uninjected (C, D, E). Expression of otx2 (C,H,M), Xanf1 and en2 (D, I, N), Xag1/N-tubulin (E, J, O). After overexpression of foxA4, the caudal diencephalic otx2 subdomain was expanded anteriorly (54%, n = 13) (H) or the telencephalic otx2 subdomain disappeared (40% n = 10) (M). The pituitary Xanf1 domain was expanded (33%, n = 15) (I). N-tubulin expression in the rostral forebrain (rectangle in E) was deleted and the Xag1 + cement and hatching glands were reduced (J) or the cement gland was absent (O) (54%, n = 24). The most severe phenotypes presented head truncations (23%, n = 13) (M,N,O). Red arrowhead, anterior limit of the caudal diencephalic otx2 subdomain; white arrowhead, posterior limit of the ventral telencephalic otx2 subdomain; vtel, ventral telencephalic otx2; cdi, caudal diencephalic otx2; mes, mesencephalic otx2; op, olfactory placode; epiph, epiphisis; ev, eye vesicle; cg, cement gland; hg, hatching gland cells; pit, pituitary anlage; is, injected side; nis, non-injected side.
doi:10.1371/journal.pone.0110559.g006 |
