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Whole-mount immunostaining of Drosophila skeletal muscle

Nature Protocols volume 8, pages 2496–2501 (2013)Cite this article

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Abstract

Skeletal muscle undergoes marked functional decay during aging in humans, but the cell biological mechanisms responsible for this process are only partly known. Age-related muscle dysfunction is also a feature of aging in the fruit fly Drosophila melanogaster. Here we describe a detailed step-by-step protocol, which takes place over 3 d, for whole-mount immunostaining of Drosophila flight muscle. The skeletal muscle is fixed and permeabilized without any tissue freezing and dehydration so that antigens are accessible for staining with appropriate antibodies and the overall tissue ultrastructure is well preserved. This technique can be used to identify age-related cellular changes driving skeletal muscle aging and for characterizing models of human muscle disease in Drosophila.

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Figure 1: Staining of Drosophila indirect flight muscle from young and old flies.
Figure 2: Staining of Drosophila jump muscle expressing GFP with the BG487-Gal4 driver.

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Acknowledgements

F.D. is supported by funding from the American Lebanese Syrian Associated Charities (ALSAC) and the Ellison Medical Foundation New Scholar in Aging award.

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  1. Department of Developmental Neurobiology, Division of Developmental Biology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Liam C Hunt & Fabio Demontis

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  1. Liam C Hunt
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  2. Fabio Demontis
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F.D. and L.C.H. wrote the manuscript.

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Correspondence to Fabio Demontis.

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The authors declare no competing financial interests.

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Hunt, L., Demontis, F. Whole-mount immunostaining of Drosophila skeletal muscle. Nat Protoc 8, 2496–2501 (2013). https://doi.org/10.1038/nprot.2013.156

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