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Fig. 7. ADMP2 loss and blood formation. (A and B) Posterior injection of 10 ng of antisense ADMP2 morpholino oligonucleotide (A-MO) into each of the two posterior blastomeres at the four cell stage inhibited the expression of globin in the posterior ventral blood islands (c-MO: control morpholino). (C) Co-injection of 10 pg of ADMP2 δ-5′ UTR RNA (total 20 pg) with A-MO restored globin expression (white arrowhead). (D–F) Anterior injection of 5 and 10 ng/blastomere of A-MO. While the 5 ng dose inhibited globin expression when injected posteriorly (E), no reduction in globin was observed at this dose anteriorly. At 10 ng (F) a strong reduction in globin expression anteriorly was observed. (G and H) The expression of SCL, an early blood marker, was also reduced by injection of 10 ng of A-MO posteriorly when examined at stage 24. (I and J) Transverse sections at the level of the posterior blood islands show that the expression of globin was decreased to a thin layer in the A-MO-injected embryos (black arrowhead in panel J compared to that in panel I). (K) In embryos co-injected with A-MO and ADMP2 δ-5′ UTR RNA, the globin-expression extended deep into the mesodermal layer. (L) Enlarged view of ventral mid-line showing globin expression in either c-MO (left) or A-MO-injected embryos. (M, N) Coinjection of 10 ng A-MO into each blastomere at four cell stage reduced globin expression in response to BMP7 RNA injection (1 ng per embryo). Note that both control (M) and A-MO-injected embryos (N) have the same DAI 0 phenotype. |
