PUBLICATION

In vivo dynamics of skeletal muscle Dystrophin in zebrafish embryos revealed by improved FRAP analysis

Authors
Bajanca, F., Gonzalez-Perez, V., Gillespie, S.J., Beley, C., Garcia, L., Theveneau, E., Sear, R.P., Hughes, S.M.
ID
ZDB-PUB-151016-12
Date
2015
Source
eLIFE   4: (Journal)
Registered Authors
Hughes, Simon M.
Keywords
Dystrophin, FRAP, binding dynamics, cell biology, developmental biology, diffusion, muscle, software, stem cells, zebrafish
MeSH Terms
  • Animals
  • Dystrophin/analysis*
  • Fluorescence Recovery After Photobleaching
  • Humans
  • Muscle, Skeletal/chemistry*
  • Muscle, Skeletal/growth & development*
  • Zebrafish/embryology*
PubMed
26459831 Full text @ Elife
Abstract
Dystrophin forms an essential link between sarcolemma and cytoskeleton, perturbation of which causes muscular dystrophy. We analysed Dystrophin binding dynamics in vivo for the first time. Within maturing fibres of host zebrafish embryos, our analysis reveals a pool of diffusible Dystrophin and complexes bound at the fibre membrane. Combining modelling, an improved FRAP methodology and direct semi-quantitative analysis of bleaching suggests the existence of two membrane-bound Dystrophin populations with widely differing bound lifetimes: a stable, tightly bound pool, and a dynamic bound pool with high turnover rate that exchanges with the cytoplasmic pool. The three populations were found consistently in human and zebrafish Dystrophins overexpressed in wild-type or dmd(ta222a/ta222a) zebrafish embryos, which lack Dystrophin, and in Gt(dmd-Citrine)(ct90a) that express endogenously-driven tagged zebrafish Dystrophin. These results lead to a new model for Dystrophin membrane association in developing muscle, and highlight our methodology as a valuable strategy for in vivo analysis of complex protein dynamics.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes