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Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry

Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry

Published on: June 6, 2011

Visualizing enveloping layer glycans during zebrafish early embryogenesis.

Jeremy M Baskin¹, Karen W Dehnert, Scott T Laughlin

¹Department of Chemistry, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.

Proceedings of the National Academy of Sciences of the United States of America

|May 22, 2010

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This study introduces a new method to visualize glycans in early zebrafish embryos, enabling the imaging of mucin-type O-glycans and sialylated glycans during gastrulation. This breakthrough allows for detailed observation of glycan dynamics in developing embryos.

Area of Science:

Developmental Biology
Chemical Biology
Glycobiology

Background:

Cell surface glycans are crucial for developmental processes.
Previous imaging methods failed to detect glycans in early zebrafish embryogenesis (first 24 hours).

Purpose of the Study:

To develop a novel approach for imaging glycans during early zebrafish embryogenesis.
To visualize mucin-type O-glycans and sialylated glycans in the enveloping layer.

Main Methods:

Microinjection of azidosugars into one-cell stage zebrafish embryos.
Detection of metabolically labeled glycans using copper-free click chemistry.
Nonmetabolic labeling of sialylated glycans for simultaneous imaging.

Main Results:

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Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry

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A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos

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Enabled imaging of mucin-type O-glycans as early as 7 hours postfertilization (gastrula stage).
Simultaneously imaged two distinct classes of glycans (mucin-type O-glycans and sialylated glycans).
Revealed rapid glycan reorganization on the second timescale, including migration to the cleavage furrow of mitotic cells.

Conclusions:

Provides a method to image glycans in the enveloping layer during early zebrafish embryogenesis.
Offers insights into glycan biosynthesis and dynamics during development.
Establishes a platform for imaging other biomolecular targets via microinjection.