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

Bajanca, F, Gonzalez-Perez, V, Gillespie, SJ, Beley, C, Garcia, L, Theveneau, E, Sear, RP and Hughes, SM (2015) In vivo dynamics of skeletal muscle Dystrophin in zebrafish embryos revealed by improved FRAP analysis ELIFE, 4, ARTN e.

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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 dmdta222a/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

Item Type: Article
Divisions : Faculty of Engineering and Physical Sciences > Physics
Authors :
AuthorsEmailORCID
Bajanca, FUNSPECIFIEDUNSPECIFIED
Gonzalez-Perez, VUNSPECIFIEDUNSPECIFIED
Gillespie, SJUNSPECIFIEDUNSPECIFIED
Beley, CUNSPECIFIEDUNSPECIFIED
Garcia, LUNSPECIFIEDUNSPECIFIED
Theveneau, EUNSPECIFIEDUNSPECIFIED
Sear, RPUNSPECIFIEDUNSPECIFIED
Hughes, SMUNSPECIFIEDUNSPECIFIED
Date : 13 October 2015
Identification Number : 10.7554/eLife.06541
Uncontrolled Keywords : Science & Technology, Life Sciences & Biomedicine, Biology, Life Sciences & Biomedicine - Other Topics, DUCHENNE MUSCULAR-DYSTROPHY, MESSENGER-RNA LOCALIZATION, DEFICIENT ZEBRAFISH, PROTEIN, ACTIN, EXPRESSION, DIFFUSION, CELLS, MEMBRANE, COMPLEX
Related URLs :
Additional Information : Copyright Bajanca et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
Depositing User : Symplectic Elements
Date Deposited : 12 Nov 2015 12:09
Last Modified : 12 Nov 2015 12:09
URI: http://epubs.surrey.ac.uk/id/eprint/809331

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