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First published online September 9, 2004
doi: 10.1242/10.1242/dev.01331
1 Gene Expression Laboratory, The Salk Institute for Biological Studies, La
Jolla, CA 92037-1099, USA
2 Unidade de Desenvolvimento, Instituto Gulbenkian de Ciencia, Rua Da quinta
Grande 6 Aptdo 14. 2780-901 Oeiras, Portugal
3 The Institute of Molecular and Cellular Biosciences, The University of Tokyo,
1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-0032, Japan
* Author for correspondence (e-mail: belmonte{at}salk.edu)
Accepted 28 June 2004
Initiation and maintenance of signaling centers is a key issue during embryonic development. The apical ectodermal ridge, a specialized epithelial structure and source of Fgf8, is a pivotal signaling center for limb outgrowth. We show that two closely related buttonhead-like zinc-finger transcription factors, Sp8 and Sp9, are expressed in the AER, and regulate Fgf8 expression and limb outgrowth. Embryological and genetic analyses have revealed that Sp8 and Sp9 are ectodermal targets of Fgf10 signaling from the mesenchyme. We also found that Wnt/ß-catenin signaling positively regulates Sp8, but not Sp9. Overexpression functional analyses in chick unveiled their role as positive regulators of Fgf8 expression. Moreover, a dominant-negative approach in chick and knockdown analysis with morpholinos in zebrafish revealed their requirement for Fgf8 expression and limb outgrowth, and further indicate that they have a coordinated action on Fgf8 expression. Our study demonstrates that Sp8 and Sp9, via Fgf8, are involved in mediating the actions of Fgf10 and Wnt/ß-catenin signaling during vertebrate limb outgrowth.
Key words: Apical ectodermal ridge, Sp transcription factor, Fgf8, Fgf10, Wnt, Vertebrate limb
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