Effect of Fibroblast Growth Factors on Outgrowth of Facial Mesenchyme

Abstract

The ectoderm is required for outgrowth of facial prominences and facial ectoderm from all facial prominences is interchangeable. Signals provided by the ectoderm may include members of the fibroblast growth factor family (FGF). In order to test whether FGFs could replace facial ectoderm and promote outgrowth, stage 24 frontonasal mass or mandibular mesenchyme was grafted to a host chick limb and a bead soaked in FGF-2 or FGF-4 was placed on top of the mesenchyme. Following 7 days of incubation, the amount of outgrowth was quantified by measuring the rods of cartilage that formed from the grafts. FGF-2 and FGF-4 stimulated an increase in length of cartilage rods in mandibular grafts compared to mandibular mesenchyme grafted without ectoderm (P< 0.05). FGF-4 stimulated a small increase in length of frontonasal mass mesenchyme (P< 0.05) and both FGFs increased the frequency of egg tooth formation in frontonasal mass mesenchyme compared to frontonasal mass mesenchyme grafted without ectoderm. FGFs can partially but not completely replace facial ectoderm since homotypic recombinations of frontonasal mass and mandibular tissues were significantly longer than mesenchyme grafts treated with FGF-soaked beads (P< 0.05). The addition of a second FGF-soaked bead did not significantly increase the length of the frontonasal mass or the mandibular mesenchyme. We have determined that FGF-2 protein is expressed in facial ectoderm and could be an endogenous signal for outgrowth. In contrast, FGF-8 transcripts are not expressed in the ectoderm covering the areas of the face that were grafted; thus, it is less likely that FGF-8 is required for outgrowth. Our results indicate that FGFs are part of an endogenous signaling pathway involved in distal outgrowth and chondrogenesis of the facial prominences.