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Imaging glycans in zebrafish embryos by metabolic labeling and bioorthogonal click chemistry

Academic Article
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Overview

authors

  • Jiang, H.
  • Feng, L.
  • Soriano del Amo, D.
  • Seidel III, R. D.
  • Marlow, F. L.
  • Wu, Peng

publication date

  • June 2011

journal

  • Journal of Visualized Experiments : JoVE  Journal

abstract

  • Imaging glycans in vivo has recently been enabled using a bioorthogonal chemical reporter strategy by treating cells or organisms with azide- or alkyne-tagged monosaccharides. The modified monosaccharides, processed by the glycan biosynthetic machinery, are incorporated into cell surface glycoconjugates. The bioorthogonal azide or alkyne tags then allow covalent conjugation with fluorescent probes for visualization, or with affinity probes for enrichment and glycoproteomic analysis. This protocol describes the procedures typically used for noninvasive imaging of fucosylated glycans in zebrafish embryos, including: 1) microinjection of one-cell stage embryos with GDP-5-alkynylfucose (GDP-FucAl), 2) labeling fucosylated glycans in the enveloping layer of zebrafish embryos with azide-conjugated fluorophores via biocompatible Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC), and 3) imaging by confocal microscopy. The method described here can be readily extended to visualize other classes of glycans, e.g. glycans containing sialic acid and N-acetylgalactosamine, in developing zebrafish and in other living organisms.

subject areas

  • Alkynes
  • Animals
  • Azides
  • Click Chemistry
  • Embryo, Nonmammalian
  • Fluorescent Dyes
  • Guanosine Diphosphate Fucose
  • Microinjections
  • Microscopy, Confocal
  • Polysaccharides
  • Zebrafish
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Research

keywords

  • Developmental Biology
  • Issue 52
  • chemical glycobiology
  • click chemistry
  • embryogenesis
  • fucosylated glycans
  • microinjection
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Identity

PubMed Central ID

  • PMC3197039

Digital Object Identifier (DOI)

  • 10.3791/2686

PubMed ID

  • 21673647
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Additional Document Info

start page

  • pii: 2686

issue

  • 52

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