ZFIN ID: ZDB-PUB-150618-3
Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line
Ruf-Zamojski, F., Trivedi, V., Fraser, S.E., Trinh, L.A.
Date: 2015
Source: PLoS One   10: e0128944 (Journal)
Registered Authors: Ruf-Zamojski, Frederique, Trinh, Le
Keywords: none
MeSH Terms:
  • Animals
  • Bacterial Proteins/genetics*
  • Bacterial Proteins/metabolism
  • Embryo, Nonmammalian
  • Gene Expression Regulation, Developmental
  • Luminescent Proteins/genetics*
  • Luminescent Proteins/metabolism
  • Membrane Proteins/genetics*
  • Membrane Proteins/metabolism
  • Microscopy, Fluorescence
  • Morphogenesis/genetics
  • Morpholinos
  • Muscle Proteins/genetics*
  • Muscle Proteins/metabolism
  • Muscle, Skeletal/cytology
  • Muscle, Skeletal/embryology
  • Muscle, Skeletal/metabolism*
  • Plasmids/chemistry
  • Plasmids/metabolism
  • RNA, Messenger/genetics*
  • RNA, Messenger/metabolism
  • Recombinant Fusion Proteins/genetics*
  • Recombinant Fusion Proteins/metabolism
  • Time-Lapse Imaging
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
PubMed: 26083378 Full text @ PLoS One
FIGURES
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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