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Myogenesis pp 153–169Cite as

Immunocytochemistry to Study Myogenesis in Zebrafish

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 798))

Abstract

During myogenesis, cells gradually transition from mesodermal precursors to myoblasts, myocytes, and then to muscle fibers. The molecular characterization of this process requires the ability to identify each of these cell types and the factors that regulate the transitions between them. The most versatile technique for assaying cell identities in situ is immunocytochemistry, because multiple independent molecular markers of differentiation can be assayed simultaneously. The zebrafish has developed into a popular model for the study of myogenesis, and immunocytochemical techniques have been critical. We have adapted existing protocols to optimize immunocytochemistry in zebrafish, and have tested many antibodies developed against mouse, chick, and frog muscle antigens for their cross-reactivity in zebrafish. Here, we present protocols for whole mount immunocytochemistry on both formaldehyde and Carnoy’s fixed embryos as well as on sectioned zebrafish tissue. We include a table of antibodies useful for experiments on the molecular biology of myogenesis in zebrafish.

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Acknowledgments

This work was supported by an NIH grant to SHD (R01 HD044929). We thank other labs for sharing protocols, and all members of the Devoto Lab, past and present, who helped to test and refine this protocol.

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Authors and Affiliations

  1. Department of Biology, Wesleyan University, Hall-Atwater Laboratories, Middletown, CT, USA

    Nathan C. Bird, Stefanie E. Windner & Stephen H. Devoto

Authors
  1. Nathan C. Bird
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  2. Stefanie E. Windner
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  3. Stephen H. Devoto
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Corresponding author

Correspondence to Stephen H. Devoto .

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Editors and Affiliations

  1. Chicago Medical School, Dept. Cell Biology & Anatomy, Rosalind Franklin Univ of Med & Sci, Green Bay Rd. 3333, North Chicago, 60064, Illinois, USA

    Joseph X. DiMario

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Bird, N.C., Windner, S.E., Devoto, S.H. (2012). Immunocytochemistry to Study Myogenesis in Zebrafish. In: DiMario, J. (eds) Myogenesis. Methods in Molecular Biology, vol 798. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-343-1_9

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  • DOI: https://doi.org/10.1007/978-1-61779-343-1_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-342-4

  • Online ISBN: 978-1-61779-343-1

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