Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line |
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| Authors: | Frederique Ruf-Zamojski Vikas Trivedi Scott E. Fraser Le A. Trinh |
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| Affiliation: | 1. California Institute of Technology, Biological Imaging Center, Beckman Institute, Division of Biology, Pasadena, California 91125, United States of America.; 2. California Institute of Technology, Department of Bioengineering, Pasadena, California 91125, United States of America.; University of Minnesota Medical School, UNITED STATES, |
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| 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. |
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