Whole-mount immunostaining of Drosophila skeletal muscle.
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.
Department of Developmental Neurobiology, Division of Developmental Biology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
Liam C
LC
Hunt
Fabio
F
Demontis
D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005838:Genotype; D049452:Green Fluorescent Proteins / Q000032:analysis; D007150:Immunohistochemistry; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000032:analysis
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins