The loss of ventral ectoderm identity correlates with the inability to form an AER in the legless hindlimb bud

We have characterized the early stages of murine hindlimb morphogenesis in the legless (lgl)mutant and non-mutant littermates. Initially the entire ventral ectoderm expresses many genetic markers characteristic of the AER (en-1, fgf-8, msx-2, dlx-2, cd44, and cx-43). Subsequently, the expression domain of most of these genes is restricted to the thickened ectoderm of the disto-ventral limb margin prior to forming an AER. In lgl, the expression of these genes is initiated but not maintained and the disto-ventral marginal ectoderm does not thicken. In contrast, Wnt7a expression is initiated and maintained in the dorsal ectoderm. The limb mesenchyme of lgl and non-mutant embryos initially expresses lmx-1b and fgf-10 uniformly. As the ventro-distal marginal ectoderm thickens, lmx-1b is progressively dorsally restricted in non-mutants but continues to be expressed ventrally in lgl hindlimb buds. These data suggest that establishment of a dorso-ventral ectodermal interface is not sufficient for AER formation and that restriction of lmx-1b to the dorsal mesenchyme is coordinately linked to AER formation.