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Fig. 1. Xrx1 expression in the proliferative region of the anterior neural plate is controlled by Hedgehog and neurogenin signaling. (A,B) Expression of Xrx1 (light blue) in relation to the expression of (A) X-Delta-1 (purple) and (B) X-ngnr-1 (purple) in stage 13 embryos; frontodorsal views. (C) Sagittal section of a stage 13 embryo showing Xrx1 expression in the deep sensorial layer of the neuroectoderm. (D,E) Stage 14 embryos injected with X-chh (D) and X-shh (E) showing ectopic expression of Xrx1 (blue); frontal views, dorsal towards the top. (F,G) Embryos injected with X-ngnr-1 displaying repression of Xrx1 (blue, F, stage 13) and ectopic expression of N-tubulin (blue, G, stage 16); frontodorsal views. The injected side of the embryos (to the right of vertical bars representing the midline) is indicated (inj). Red staining represents expression of co-injected lacZ lineage tracer. d, deep neuroectodermal layer; s, superficial neuroectodermal layer. |
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Fig. 6. (A-D) Embryos injected with Xrx1 were treated with HUA at stage 10.5 and the expression of Zic2 (A), Xhairy2 (B), X-ngnr-1 (C) and p27Xic1 (D) was analyzed at stage 13. (A,B) Black brackets indicate the anterior expression domains in the uninjected side; white brackets indicate the corresponding enlarged domains in the injected side. (C,D) Black arrowheads indicate anterior expression domains in the uninjected side, white arrowheads indicate the absence of this expression domain in the injected side. (E-I) HUA treatment dramatically reduced anti-phosphoH3 staining (E,F; stage 16) as well as Xoptx2 ability of expanding Xrx1 (Zuber et al., 1999) (G,H; stage 18). This treatment also results in a reduction of the optic vesicle size (I; stage 26; Co, control untreated embryo). (A-D,G,H) Frontal views, dorsal towards the top; (E,F,I) lateral views, anterior towards the left. Red staining in A-D and turquoise staining in G,H represent expression of co-injected lacZ lineage tracer. The injected side of the embryos (to the right of vertical bars representing the midline) is indicated (inj). |
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Fig. 8. Effects of Xrx1 loss of function on genes regulating anterior neurogenesis. (A-C) Phenotypes of stage 41 embryos injected with MoXrx1 (A), MoXrx1 and Xrx1 (B), and control morpholino oligo (C). (F,J,K,M,N,Q) Embryos injected with MoXrx1 in both dorsoanimal blastomeres at the eight-cell stage and analyzed at stage 14 (F,K,M,N,Q) and stage 18 (J). (D,G,H,L,P) Control uninjected embryos analyzed at stage 14 (D,G,L,P) and stage 18 (H). (E,I) Embryos bilaterally injected with Xrx1-EnR analyzed at stage 14 (E) and stage 18 (I). (O) Stage 14 embryo treated with HUA. (R,S,V) Stage 14 embryos bilaterally injected with Xhairy2 (R), MoXrx1 and Xhairy2 (S), and MoXrx1 and XBF-1 (V). (T,U) Dorsal (T) and frontal (U) views of a stage 14 embryo injected with XBF-1. Red staining represents co-injected lacZ. Black brackets indicate the size of the anterior expression domain in the uninjected embryo (P); white brackets indicate the size of the corresponding domain in the injected embryo (Q). Arrows in D,E,F,S,V indicate the anterior boundaries of X-ngnr-1 expression; the arrow in T indicates X-ngnr-1 ectopic expression. Black arrowheads in I,J indicate the continuous anterior extension of X-ngnr-1 expression. White arrowheads in U and R indicate the repressed anterior expression domain of X-ngnr-1. |
