title: Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye.
abstract: The zebrafish model is an emerging system for the study of neuromuscular disorders. In the study of neuromuscular diseases, the integrity of the muscle membrane is a critical disease determinant. To date, numerous neuromuscular conditions display degenerating muscle fibers with abnormal membrane integrity; this is most commonly observed in muscular dystrophies. Evans Blue Dye (EBD) is a vital, cell permeable dye that is rapidly taken into degenerating, damaged, or apoptotic cells; in contrast, it is not taken up by cells with an intact membrane. EBD injection is commonly employed to ascertain muscle integrity in mouse models of neuromuscular diseases. However, such EBD experiments require muscle dissection and/or sectioning prior to analysis. In contrast, EBD uptake in zebrafish is visualized in live, intact preparations. Here, we demonstrate a simple and straightforward methodology for performing EBD injections and analysis in live zebrafish. In addition, we demonstrate a co-injection strategy to increase efficacy of EBD analysis. Overall, this video article provides an outline to perform EBD injection and characterization in zebrafish models of neuromuscular disease.
authors.affiliation: Program in Genetics & Genome Biology, The Hospital for Sick Children; Department of Molecular Genetics, The University of Toronto.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Smith
authors.affiliation: Program in Genomics of Differentiation, Eunice Kennedy Shriver National Institute of Child Health and Human Development; Departments of Pediatrics and Neurology, University of Michigan.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Horstick
authors.affiliation: Program in Genetics & Genome Biology, The Hospital for Sick Children; Department of Molecular Genetics, The University of Toronto.
authors.forename: Ann E
authors.initials: AE
authors.lastname: Davidson
authors.affiliation: Program in Genetics & Genome Biology, The Hospital for Sick Children; Department of Molecular Genetics, The University of Toronto; Departments of Pediatrics and Neurology, University of Michigan; james.dowling@sickkids.ca.
authors.forename: James
authors.initials: J
authors.lastname: Dowling