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
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • Microscopy, Fluorescence
  • Gene Expression Regulation, Developmental
  • Animals
  • Morpholinos
  • Time-Lapse Imaging
  • RNA, Messenger/genetics*
  • RNA, Messenger/metabolism
  • Membrane Proteins/genetics*
  • Membrane Proteins/metabolism
  • Morphogenesis/genetics
  • Recombinant Fusion Proteins/genetics*
  • Recombinant Fusion Proteins/metabolism
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
  • Muscle Proteins/genetics*
  • Muscle Proteins/metabolism
  • Luminescent Proteins/genetics*
  • Luminescent Proteins/metabolism
  • Bacterial Proteins/genetics*
  • Bacterial Proteins/metabolism
  • Muscle, Skeletal/cytology
  • Muscle, Skeletal/embryology
  • Muscle, Skeletal/metabolism*
  • Plasmids/chemistry
  • Plasmids/metabolism
  • Embryo, Nonmammalian
(all 29)
PubMed
26083378 Full text @ PLoS One
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
Figures
Figure Gallery (9 images)
Show all Figures
Expression
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay Figure
Citrinedmdct90aGtstandard conditionsDay 6IFL
Citrinedmdct90aGtstandard conditionsPrim-15IHC
Citrinedmdct90aGtstandard conditionsLong-pecISH
Citrinedmdct90aGtstandard conditions14-19 somitesIHC
Citrinedmdct90aGtstandard conditionsPrim-5IHC
Citrinedmdct90aGtcontrolPrim-15IFL
Citrinedmdct90aGtstandard conditions26+ somitesIFL
Citrinedmdct90aGtstandard conditions20-25 somitesIFL
Citrinedmdct90aGtcontrolPrim-15 to Protruding-mouthIFL
Citrinedmdct90aGtheat shockPrim-15IFL
1 - 10 of 35
Show
Phenotype
Fish Conditions Stage Phenotype Figure
dmdct90aGtheat shockPrim-15
1 - 1 of 1
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
ct90aGtTransgenic Insertion
1 - 1 of 1
Show
Human Disease / Model
No data available
Sequence Targeting Reagents
Target Reagent Reagent Type
dmdMO1-dmdMRPHLNO
1 - 1 of 1
Show
Fish
Fish
dmdct90aGt/ct90aGt
dmdct90aGt/+
dmdct90aGt
dmdct90aGt + MO1-dmd
WT
WT + MO1-dmd
1 - 6 of 6
Show
Antibodies
Orthology
No data available
Engineered Foreign Genes
Marker Marker Type Name
CitrineEFGCitrine
mCherryEFGmCherry
1 - 2 of 2
Show
Mapping
No data available
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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.