Visualizing enveloping layer glycans during zebrafish early embryogenesis
- 20 May 2010
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 107 (23), 10360-10365
- https://doi.org/10.1073/pnas.0912081107
Abstract
Developmental events can be monitored at the cellular and molecular levels by using noninvasive imaging techniques. Among the biomolecules that might be targeted for imaging analysis, glycans occupy a privileged position by virtue of their primary location on the cell surface. We previously described a chemical method to image glycans during zebrafish larval development; however, we were unable to detect glycans during the first 24 hours of embryogenesis, a very dynamic period in development. Here we report an approach to the imaging of glycans that enables their visualization in the enveloping layer during the early stages of zebrafish embryogenesis. We microinjected embryos with azidosugars at the one-cell stage, allowed the zebrafish to develop, and detected the metabolically labeled glycans with copper-free click chemistry. Mucin-type O-glycans could be imaged as early as 7 hours postfertilization, during the gastrula stage of development. Additionally, we used a nonmetabolic approach to label sialylated glycans with an independent chemistry, enabling the simultaneous imaging of these two distinct classes of glycans. Imaging analysis of glycan trafficking revealed dramatic reorganization of glycans on the second time scale, including rapid migration to the cleavage furrow of mitotic cells. These studies yield insight into the biosynthesis and dynamics of glycans in the enveloping layer during embryogenesis and provide a platform for imaging other biomolecular targets by microinjection of appropriately functionalized biosynthetic precursors.Keywords
This publication has 36 references indexed in Scilit:
- In Toto Imaging of Embryogenesis with Confocal Time-Lapse MicroscopyPublished by Springer Science and Business Media LLC ,2009
- High-efficiency labeling of sialylated glycoproteins on living cellsNature Methods, 2009
- Imaging the glycomeProceedings of the National Academy of Sciences of the United States of America, 2009
- Dynamic Analyses of Drosophila Gastrulation Provide Insights into Collective Cell MigrationScience, 2008
- Reconstruction of Zebrafish Early Embryonic Development by Scanned Light Sheet MicroscopyScience, 2008
- Sialic acids in human health and diseaseTrends in Molecular Medicine, 2008
- In Vivo Imaging of Membrane-Associated Glycans in Developing ZebrafishScience, 2008
- Copper-free click chemistry for dynamic in vivo imagingProceedings of the National Academy of Sciences of the United States of America, 2007
- Imaging Cell Surface Glycans with Bioorthogonal Chemical ReportersJournal of the American Chemical Society, 2007
- Cell lineage of zebrafish blastomeresDevelopmental Biology, 1985