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Figure 6


Fig. 6. Regulation of Lmx1b expression by ß-catenin activity. (A-B'') Expression of Wnt7a in the dorsal ectoderm (A-A'') and En1 in the ventral ectoderm (B-B'') at E10.5 in wild-type (A,B), ß-cat{Delta}Prx1/- (A',B'), and ß-cat{Delta}ex3Prx1/+ (A'',B'') fore- and hindlimbs (n=4). (C,C') Dorsal view of E9.5 embryos showing uniform expression of Lmx1b in wild type (C) and downregulation in ß-cat{Delta}Prx1/- (C', n=6) forelimbs. (D,D') Lateral view of E9.5 embryos, showing restricted expression of Lmx1b in the dorsal mesenchyme (arrow) in wild type (D), which expands into the ventral mesenchyme (arrow) in ß-cat{Delta}ex3Prx1/+ (D') forelimbs. (E,E') Lmx1b expression at E11.5 in wild-type (E) and ß-cat{Delta}Prx1/- (E') forelimbs. (F-K) Section in situ hybridizations. (F-F'') At E10.5, Lmx1b expression is dorsally restricted in wild-type forelimbs (F; delineated by a line running perpendicularly through the AER, red arrow), is reduced and lost in the distal limb mesenchyme adjoining the dorsal ectoderm in ß-cat{Delta}ex3Prx1/+ (F'; white arrow), but is expanded into the ventral mesenchyme in ß-cat{Delta}ex3Prx1/+ limbs (F''; note, expression is excluded from the ventral-most regions; black arrow; n=5). (G-G'') Distribution of ß-catenin protein in wild-type (G) and two different E11.5 ß-cat{Delta}ex3Prx1/+ (G',G'') hindlimbs at E11.5. (G-J'') Alternating sections of the same E11.5 wild-type (G-J) and ß-cat{Delta}ex3Prx1/+ (G'-J') hindlimbs. (H,H') Lmx1b expression in wild type (H) and ectopic expression in gof limbs (H'). (I,I') Lef1 expression in wild type (I) and ectopic expression in gof limbs (I'). (J,J') Tcf1 expression in wild type (J) and ectopic expression in gof limbs (J'). (H'',I'',K) Fluorescent double in situ hybridization on an E11.5 ß-cat{Delta}ex3Prx1/+ hindlimb section (same limb as is shown in G''), showing that Lmx1b- (H'') and Lef1 (I'')-positive regions overlap in the distal limb bud (K: Lef1 green; Lmx1b red). Embryos are orientated anterior to the right. Limb bud sections are dorsal up, distal to the right.





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