PUBLICATION
Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line
- Authors
- Ruf-Zamojski, F., Trivedi, V., Fraser, S.E., Trinh, L.A.
- ID
- ZDB-PUB-150618-3
- 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
Citation
Ruf-Zamojski, F., Trivedi, V., Fraser, S.E., Trinh, L.A. (2015) Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line. PLoS One. 10:e0128944.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping