Essential role of heparan sulfate 2-O-sulfotransferase in chick limb bud patterning and development

J Biol Chem. 2007 Jul 6;282(27):19589-97. doi: 10.1074/jbc.M610707200. Epub 2007 May 10.


The interactions of heparan sulfate (HS) with heparin-binding growth factors, such as fibroblast growth factors (FGFs), depend greatly on the chain structures. O-Sulfations at various positions on the chain are major factors determining HS structure; therefore, O-sulfation patterns may play a crucial role in controlling the developmental and morphogenetic processes of various tissues and organs by spatiotemporally regulating the activities of heparin-binding growth factors. In a previous study, we found that HS-2-O-sulfotransferase is strongly expressed throughout the mesoderm of chick limb buds during the early stages of development. Here we show that inhibition of HS-2-O-sulfotransferase in the prospective limb region by small inhibitory RNA resulted in the truncation of limb buds and reduced Fgf-8 expression in the apical ectodermal ridge. The treatment also reduced Fgf-10 expression in the mesenchyme. Moreover 2-O-sulfated HS, normally abundant in the basement membranes and mesoderm under ectoderm in limb buds, was significantly reduced in the treated buds. Phosphorylation levels of ERK and Akt were up-regulated in such truncated buds. Thus, we have shown for the first time that 2-O-sulfation of HS is essential for the FGF signaling required for limb bud development and outgrowth.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Body Patterning / physiology*
  • Chick Embryo
  • Ectoderm / cytology
  • Ectoderm / enzymology
  • Enzyme Activation / physiology
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Fibroblast Growth Factors / metabolism
  • Gene Expression Regulation, Developmental / drug effects
  • Gene Expression Regulation, Developmental / physiology*
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Enzymologic / physiology*
  • Heparitin Sulfate / metabolism*
  • Limb Buds / cytology
  • Limb Buds / embryology
  • Limb Buds / enzymology*
  • Mesoderm / cytology
  • Mesoderm / enzymology
  • Phosphorylation
  • Protein Binding / physiology
  • Protein Processing, Post-Translational / drug effects
  • Protein Processing, Post-Translational / physiology*
  • Proto-Oncogene Proteins c-akt / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology
  • Sulfotransferases / antagonists & inhibitors
  • Sulfotransferases / biosynthesis*
  • Sulfotransferases / genetics


  • RNA, Small Interfering
  • Fibroblast Growth Factors
  • Heparitin Sulfate
  • Proto-Oncogene Proteins c-akt
  • Extracellular Signal-Regulated MAP Kinases
  • Sulfotransferases
  • heparan-sulfate 2-sulfotransferase