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  • Title: FGFR2 signaling in normal and limbless chick limb buds.
    Author: Lizarraga G, Ferrari D, Kalinowski M, Ohuchi H, Noji S, Kosher RA, Dealy CN.
    Journal: Dev Genet; 1999; 25(4):331-8. PubMed ID: 10570465.
    Abstract:
    FGF10 and FGF8, which are reciprocally expressed by the mesoderm and AER of the developing limb bud, have been implicated in limb initiation, outgrowth, and patterning. FGF10 and FGF8 signal through the FGFR2b and FGFR2c alternative splice isoforms, respectively [Ornitz DM, et al. 1996. J Biol Chem 271:15292-15297; Igarashi M, et al. 1998. J Biol Chem 273:13230-13235]. A paracrine signaling loop model has been proposed whereby FGF10 expressed by limb mesoderm signals via ectodermally restricted FGFR2b to regulate FGF8 expression by the apical ectoderm; in turn, FGF8 signals via mesodermally restricted FGFR2c to maintain FGF10 expression [Ohuchi H, et al. 1997. Development 124:2235-2244; Xu X, et al. 1998. Development 125:753-765]. To explore this model, we have examined FGFR2b and FGFR2c mRNA expression, using isoform-specific probes during the early stages of development of the chick limb when limb initiation, AER induction, and outgrowth are occurring. We have found that FGFR2b is expressed by limb ectoderm, including the AER, consistent with paracrine signaling of FGF10. By contrast, FGFR2c is expressed by both mesoderm and ectoderm, indicating that FGF8 has the potential to function in an autocrine as well as paracrine fashion. Indeed, as the limb grows out in response to the AER, FGFR2c expression attenuates in the mesoderm of the progress zone, but is maintained in the AER itself, arguing against exclusive paracrine signaling of FGF8 during limb outgrowth. We also report that transcripts for FGF10, FGFR2b, and FGFR2c are expressed normally in the limb buds of limbless mutant embryos, which fail to form an AER and do not express FGF8. Furthermore, we detect no mutations in exons specific for the FGFR2c or FGFR2b isoforms in limbless embryos. Since gene targeting has shown that expression of FGF8 in limb ectoderm depends on FGF10 [Min H, et al. 1998. Genes Dev 12:3156-3161; Sekine K, et al. 1999. Nature Genet 21:138-141], these results indicate that the product of the limbless gene is required for FGF10 to induce expression of FGF8.
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