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FIGURE 7. Decreased retinoid signaling in rdhe2 morphants. A, rdhe2- or control-MO (155 ng) mixed with the lineage tracer were injected into one dorsal blastomere at four-cell stage. The injected side is indicated by an arrow. D, rdhe2- or control-MO (50 ng/embryo) was injected into two dorsal blastomers. D, embryos were treated with 0.01 or 0.1 μM RA where specified. A, whole mount in situ hybridization with antisense RNA probes was performed for the following markers (stages and frequencies of observed phenotypes in parentheses): A, otx2 and krox20 (st. 20, 31/77); B, en2 (st. 189, 42/74); C and D, hoxd4 (st. 189, 30/46); E, sox3 (st. 134, 10/12); F, shh (st.14, 23/28); G, myod (st. 302, 15/42). Arrows in G indicate irregular somites as revealed by myod staining. H, tissues of rdhe2 and control morphants were incubated over the monolayer of F9-RARE-lacZ cells; β-galactosidase activity was revealed with X-gal substrate. The results are representative of three independent experiments. |