Summary
Myoblast fusion in Drosophila has become a powerful genetic system with which to unravel the mechanisms underlying cell fusion. The identification of important components of myoblast fusion by genetic analysis has led to a molecular pathway toward our understanding of this cellular process. In addition to the application of immunohistochemistry and live imaging techniques to visualize myoblast fusion at the light microscopic level, ultrastructural analysis using electron microscopy remains an indispensable tool to reveal fusion intermediates and specific membrane events at sites of fusion. In this chapter, we describe conventional chemical fixation and high-pressure freezing/freeze substitution methods for visualizing fusion intermediates during Drosophila myoblast fusion. Furthermore, we describe an immunoelectron microscopic method for localizing specific proteins relative to the fusion apparatus.
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Acknowledgments
We thank Michael Delannoy, Richard Fetter, Kent McDonald, and Guofeng Zhang for valuable advice on conventional chemical fixation and high-pressure freezing/freeze substitution; Ulrich Tepass for immunoelectron microscopy; and members of Chen's laboratory for comments. Work in Chen's laboratory is supported by the NIH, the Packard Foundation, and the Searle Scholars Program.
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Zhang, S., Chen, E.H. (2008). Ultrastructural Analysis of Myoblast Fusion in Drosophila . In: Chen, E.H. (eds) Cell Fusion. Methods in Molecular Biology™, vol 475. Humana Press. https://doi.org/10.1007/978-1-59745-250-2_16
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DOI: https://doi.org/10.1007/978-1-59745-250-2_16
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