Effect of FGF on Gene Expression in Chick Limb Bud Cells in Vivo and in Vitro

Abstract

Fibroblast growth factors (FGFs) are central to signaling in the developing limb. FGF-2 and FGF-4 can substitute for the apical ectodermal ridge to maintain both limb bud outgrowth and polarizing region signaling. Here, we have repeated and extended previous studies and investigated local effects of the apical ectodermal ridge on gene expression of Msx-1, 5′ members of the HoxD complex, and Bmp-2 in the limb bud mesenchyme and tested whether members of the FGF family can substitute for the ridge to maintain their expression patterns. We found that expression of Msx-1, Hoxd-13, and Bmp-2 in posterior limb bud mesenchyme is dependent on a local signal from the apical ectodermal ridge. When the apical ectodermal ridge of young chick wing buds is removed, or when posterior cells are taken from the bud and placed in culture, expression of Msx-1, Hoxd-13, and Bmp-2 is not detectable in posterior mesenchymal cells. Local application of FGF-soaked beads to posterior limb mesenchyme following ridge removal or addition of FGF to cultured cells maintains expression of Msx-1, Hoxd-13, and Bmp-2. In contrast, expression of Hoxd-11 in posterior mesenchyme appears to be stable in the absence of either the apical ectodermal ridge or FGF. Expression of Msx-1 in anterior and apical cells is also locally maintained by the apical ectodermal ridge and this effect can be reproduced by local application of FGF. Furthermore, the addition of FGF to cultured anterior limb bud cells maintains their ability to respond to positional cues when grafted back into limb buds.