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

Hao Jiang, Lei Feng, David Soriano del Amo, Ronald D. Seidel, Florence Marlow, Peng Wu

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number52
StatePublished - 2011
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)

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    Jiang, H., Feng, L., Soriano del Amo, D., Seidel, R. D., Marlow, F., & Wu, P. (2011). Imaging glycans in zebrafish embryos by metabolic labeling and bioorthogonal click chemistry. Journal of visualized experiments : JoVE, (52).