PUBLICATION

Analysis of Zebrafish Larvae Skeletal Muscle Integrity with Evans Blue Dye

Authors
Smith, S.J., Horstick, E.J., Davidson, A.E., Dowling, J.
ID
ZDB-PUB-151210-5
Date
2015
Source
Journal of visualized experiments : JoVE   (105): (Journal)
Registered Authors
Horstick, Eric
Keywords
Developmental Biology, Issue 105, congenital muscular dystrophies, Evans Blue Dye, zebrafish, sarcolemma integrity, myopathy, Duchenne muscular dystrophy, dystroglycanopathies
MeSH Terms
none
PubMed
26649573 Full text @ J. Vis. Exp.
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.
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