Roles of FGFR3 during morphogenesis of Meckel's cartilage and mandibular bones.

To address the functions of FGFR2 and FGFR3 signaling during mandibular skeletogenesis, we over-expressed in the developing chick mandible, replication-competent retroviruses carrying truncated FGFR2c or FGFR3c that function as dominant negative receptors (RCAS-dnFGFR2 and RCAS-dnFGFR3). Injection of RCAS-dnFGFR3 between HH15 and 20 led to reduced proliferation, increased apoptosis, and decreased differentiation of chondroblasts in Meckel's cartilage. These changes resulted in the formation of a hypoplastic mandibular process and truncated Meckel's cartilage. This treatment also affected the proliferation and survival of osteoprogenitor cells in osteogenic condensations, leading to the absence of five mandibular bones on the injected side. Injection of RCAS-dnFGFR2 between HH15 and 20 or RCAS-dnFGFR3 at HH26 did not affect the morphogenesis of Meckel's cartilage but resulted in truncations of the mandibular bones. RCAS-dnFGFR3 affected the proliferation and survival of the cells within the periosteum and osteoblasts. Together these results demonstrate that FGFR3 signaling is required for the elongation of Meckel's cartilage and FGFR2 and FGFR3 have roles during intramembranous ossification of mandibular bones.

FGF signaling in mandibular skeletogenesis.

OBJECTIVE: To examine the functions of FGF/FGFR signaling during mandibular skeletogenesis in ovo. DESIGN: We examined the effects of inhibition of FGF signaling during mandibular skeletogenesis by overexpressing replication-competent RCAS virus encoding a truncated form of FGFR3 in the chicken mandibular process between stages 17 and 26. RESULTS: Injection of RCAS-dnFGFR3 into the developing mandible resulted in abnormalities in a stage- and region-dependent manner. Injection at early stages of development resulted in the truncation of Meckel's cartilage, severely reduced outgrowth of the mandibular process and absence of five of the mandibular bones. Injection at later stages did not affect the outgrowth of the mandibular process and Meckel's cartilage but resulted in abnormalities in mandibular osteogenesis in a region-specific manner. The bones in the more caudal region were frequently truncated whereas bones in the more rostral regions such as dentary and splenial bones were frequently absent. CONCLUSION: Together these experiments have revealed essential roles for FGF/FGFR signaling in the elongation of Meckel's cartilage, development of osteogenic condensations and appositional growth of mandibular bones.

Tissue-specific expression of Fgfr2b and Fgfr2c isoforms, Fgf10 and Fgf9 in the developing chick mandible.

Experimental evidence has demonstrated the importance of FGF signalling in morphogenesis of the mandibular processes. FGFs transmit their signals through four tyrosine kinase transmembrane receptors (FGFRs). Alternative splicing in FGFRs including FGFR2 generates different isoforms that exhibit different ligand-specificities, exclusive tissue distributions and specific biological functions. Despite extensive information regarding the isoform-specific patterns of expression Fgfr2c and Fgfr2b during morphogenesis of many organs, a comparative analysis of these specific isoforms in the chick mandible has not been reported. To better understand the function of FGFR2 in mandibular morphogenesis, we have analysed the expression Fgfr2b, Fgfr2c and their putative ligands Fgf10 and Fgf9, in the developing chick mandibular processes by in situ hybridisation and RT-PCR. Our observations show that Fgfr2b was primarily expressed in the mandibular epithelium while Fgfr2c was expressed in the mandibular mesenchyme including Meckel's cartilage. Fgf9 and Fgf10 were expressed in a variety of craniofacial regions including the mandibular epithelium and mesenchyme respectively. The temporal and spatial distributions of Fgfr2b, Fgfr2c, Fgf10 and Fgf9 in the developing mandible reported in this study make them attractive candidates for involvement in epithelial-mesenchymal signalling interactions that are known to be necessary for proper mandibular outgrowth and morphogenesis.