An important developmental role for oligosaccharides during early embryogenesis of cyprinid fish

  1. Jeroen Bakkers*,
  2. Carlos E. Semino,
  3. Henri Stroband,
  4. Jan W. Kijne*,
  5. Phillips W. Robbins, and
  6. Herman P. Spaink*,§
  1. *Institute of Molecular Plant Sciences, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands; Center for Cancer Research, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge MA 02139-4307; and Department of Animal Morphology and Cell Biology, Wageningen Agricultural University, Marijkeweg 40, 6709 PG Wageningen, The Netherlands

Abstract

Derivatives of chitin oligosaccharides have been shown to play a role in plant organogenesis at nanomolar concentrations. Here we present data which indicate that chitin oligosaccharides are important for embryogenesis in vertebrates. We characterize chitin oligosaccharides synthesized in vitro by zebrafish and carp embryos in the late gastrulation stage by incorporation of radiolabeled N-acetyl-d-[U14C]glucosamine and by HPLC in combination with enzymatic conversion using the Bradyrhizobium NodZ α-1,6-fucosyltransferase and chitinases. A rapid and sensitive bioassay for chitin oligosaccharides was also used employing suspension-cultured plant cells of Catharanthus roseus. We show that chitin oligosaccharide synthase activity is apparent only during late gastrulation and can be inhibited by antiserum raised against the Xenopus DG42 protein. The DG42 protein, a glycosyltransferase, is transiently expressed between midblastula and neurulation in Xenopus and zebrafish embryogenesis. Microinjection of the DG42 antiserum or the Bradyrhizobium NodZ enzyme in fertilized eggs of zebrafish led to severe defects in trunk and tail development.

Footnotes

  • § To whom reprint requests should be addressed.

  • Phillips W. Robbins

  • ABBREVIATIONS:
    GDP-[U-14C]fucose,
    guanosine 5′-diphospho-β-[U-14C]-l-fucose;
    GlcNAc,
    N-acetylglucosamine;
    LCO,
    lipo-chitin oligosaccharide;
    Nod,
    nodulation;
    UDP,
    uridine diphosphate
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