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Development 128, 2175-2186 (2001)
© 2001 The Company of Biologists Limited

sprouty4 acts in vivo as a feedback-induced antagonist of FGF signaling in zebrafish

Maximilian Fürthauer1, Frank Reifers2, Michael Brand2, Bernard Thisse1 and Christine Thisse1,*

1 Institut de Génétique et Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, BP 163, 67404 Illkirch cedex, CU de Strasbourg, France
2 Max Planck Institute for Cell Biology and Genetics, Dresden. c/o Department of Neurobiology University of Heidelberg Im Neuenheimer Feld 364 D-69120 Heidelberg, Germany

*Author for correspondence (e-mail: thisse{at}igbmc.u-strasbg.fr)

Accepted March 26, 2001

In looking for novel factors involved in the regulation of the fibroblast growth factor (FGF) signaling pathway, we have isolated a zebrafish sprouty4 gene, based on its extensive similarities with the expression patterns of both fgf8 and fgf3. Through gain- and loss-of-function experiments, we demonstrate that Fgf8 and Fgf3 act in vivo to induce the expression of Spry4, which in turn can inhibit activity of these growth factors. When overexpressed at low doses, Spry4 induces loss of cerebellum and reduction in size of the otic vesicle, thereby mimicking the fgf8/acerebellar mutant phenotype. Injections of high doses of Spry4 cause ventralization of the embryo, an opposite phenotype to the dorsalisation induced by overexpression of Fgf8 or Fgf3. Conversely we have shown that inhibition of Spry4 function through injection of antisense morpholino oligonucleotide leads to a weak dorsalization of the embryo, the phenotype expected for an upregulation of Fgf8 or Fgf3 signaling pathway. Finally, we show that Spry4 interferes with FGF signaling downstream of the FGF receptor 1 (FGFR1). In addition, our analysis reveals that signaling through FGFR1/Ras/mitogen-activated protein kinase pathway is involved, not in mesoderm induction, but in the control of the dorsoventral patterning via the regulation of bone morphogenetic protein (BMP) expression.

Key words: BMP, Sprouty4, FGF3, FGF8, FGFR, ERK, MAPK, Ras, Morpholino, Zebrafish




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© The Company of Biologists Ltd 2001