Abstract
Dystrophin (Dmd) is a structural protein that links the extracellular matrix to actin filaments in muscle fibers and is required for the maintenance of muscles integrity. Mutations in Dmd lead to muscular dystrophies in humans and other vertebrates. Here, we report the characterization of a zebrafish gene trap line that fluorescently labels the endogenous Dmd protein (Dmd-citrine, Gt(dmd-citrine) ct90a). We show that the Dmd-citrine line recapitulates endogenous dmd transcript expression and Dmd protein localization. Using this Dmd-citrine line, we follow Dmd localization to the myosepta in real-time using time-lapse microscopy, and find that the accumulation of Dmd protein at the transverse myosepta coincides with the onset of myotome formation, a critical stage in muscle maturation. We observed that Dmd protein localizes specifically to the myosepta prior to dmd mRNA localization. Additionally, we demonstrate that the Dmd-citrine line can be used to assess muscular dystrophy following both genetic and physical disruptions of the muscle.
Highlights
Dystrophin (Dmd) is an essential structural protein in the Dystrophin Associated Protein Complex (DAPC) of skeletal muscles that links the extracellular matrix to the cytoskeletal actin filaments of the myofibril [1, 2]
Assessing the expression of dmd in one hour developmental time intervals starting at 19.5 hpf, we found that dmd transcript switches from a non-localized diffused pattern in the cytoplasm of the muscle cells at 21.5hpf, to a restricted pattern at the transverse myosepta at 22.5 hpf (Fig 4a–4d), where it remains throughout development
We describe a zebrafish gene trap line, Gt(dmd-citrine)ct90a that expresses a Dmd-citrine protein fusion allowing the dynamic study of muscular dystrophy
Summary
Dystrophin (Dmd) is an essential structural protein in the Dystrophin Associated Protein Complex (DAPC) of skeletal muscles that links the extracellular matrix to the cytoskeletal actin filaments of the myofibril [1, 2]. Mutations in components of the DAPC result in muscular dystrophy, a condition characterized by progressive muscle weakness and degeneration. Dmd mutations are the cause of approximately 90% of all human forms of muscular dystrophies. DMD is the gene that is mutated in most cases of Becker Muscular Dystrophy and Duchenne Muscular Dystrophy (BMD and DMD, respectively) [3,4,5,6]. PLOS ONE | DOI:10.1371/journal.pone.0128944 June 17, 2015
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