title: Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line.
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.
authors.affiliation: California Institute of Technology, Biological Imaging Center, Beckman Institute, Division of Biology, Pasadena, California 91125, United States of America.
authors.forename: Frederique
authors.initials: F
authors.lastname: Ruf-Zamojski
authors.affiliation: California Institute of Technology, Department of Bioengineering, Pasadena, California 91125, United States of America.
authors.forename: Vikas
authors.initials: V
authors.lastname: Trivedi
authors.affiliation: California Institute of Technology, Biological Imaging Center, Beckman Institute, Division of Biology, Pasadena, California 91125, United States of America.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Fraser
authors.affiliation: California Institute of Technology, Biological Imaging Center, Beckman Institute, Division of Biology, Pasadena, California 91125, United States of America.
authors.forename: Le A
authors.initials: le A
authors.lastname: Trinh
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009024:Morphogenesis / Q000235:genetics; D060172:Morpholinos; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D059008:Time-Lapse Imaging; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D060172:Morpholinos; D009124:Muscle Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029961:Zebrafish Proteins; C434479:citrine protein, bacteria; C435224:dmd protein, zebrafish