In vivo myosin step-size from zebrafish skeletal muscle

Thomas P Burghardt, Katalin Ajtai, Xiaojing Sun, Naoko Takubo, Yihua Wang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Muscle myosins transduce ATP free energy into actin displacement to power contraction. In vivo, myosin side chains are modified post-translationally under native conditions, potentially impacting function. Single myosin detection provides the 'bottom-up' myosin characterization probing basic mechanisms without ambiguities inherent to ensemble observation. Macroscopic muscle physiological experimentation provides the definitive 'top-down' phenotype characterizations that are the concerns in translational medicine. In vivo single myosin detection in muscle from zebrafish embryo models for human muscle fulfils ambitions for both bottom-up and topdown experimentation. A photoactivatable green fluorescent protein (GFP)-tagged myosin light chain expressed in transgenic zebrafish skeletal muscle specifically modifies the myosin lever-arm. Strychnine induces the simultaneous contraction of the bilateral tail muscles in a live embryo, causing them to be isometric while active. Highly inclined thin illumination excites the GFP tag of single lever-arms and its super-resolution orientation is measured from an active isometric muscle over a time sequence covering many transduction cycles. Consecutive frame lever-arm angular displacement converts to step-size by its product with the estimated lever-arm length. About 17% of the active myosin steps that fall between 2 and 7 nm are implicated as powerstrokes because they are beyond displacements detected from either relaxed or ATP-depleted (rigor) muscle.

Original languageEnglish (US)
Article number160075
JournalOpen Biology
Volume6
Issue number5
DOIs
StatePublished - May 1 2016

Fingerprint

Zebrafish
Myosins
Muscle
Skeletal Muscle
Muscles
Green Fluorescent Proteins
Embryonic Structures
Adenosine Triphosphate
Strychnine
Myosin Light Chains
Translational Medical Research
Lighting
Tail
Actins
Free energy
Medicine
Byproducts
Observation
Phenotype

Keywords

  • Highly inclined thin illumination
  • Single myosin detection in vivo
  • Strychnine induced contraction
  • Transgenic zebrafish skeletal muscle
  • Zebrafish skeletal myosin powerstroke
  • Zebrafish skeletal myosin step-size

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology

Cite this

Burghardt, T. P., Ajtai, K., Sun, X., Takubo, N., & Wang, Y. (2016). In vivo myosin step-size from zebrafish skeletal muscle. Open Biology, 6(5), [160075]. https://doi.org/10.1098/rsob.160075

In vivo myosin step-size from zebrafish skeletal muscle. / Burghardt, Thomas P; Ajtai, Katalin; Sun, Xiaojing; Takubo, Naoko; Wang, Yihua.

In: Open Biology, Vol. 6, No. 5, 160075, 01.05.2016.

Research output: Contribution to journalArticle

Burghardt, TP, Ajtai, K, Sun, X, Takubo, N & Wang, Y 2016, 'In vivo myosin step-size from zebrafish skeletal muscle', Open Biology, vol. 6, no. 5, 160075. https://doi.org/10.1098/rsob.160075
Burghardt TP, Ajtai K, Sun X, Takubo N, Wang Y. In vivo myosin step-size from zebrafish skeletal muscle. Open Biology. 2016 May 1;6(5). 160075. https://doi.org/10.1098/rsob.160075
Burghardt, Thomas P ; Ajtai, Katalin ; Sun, Xiaojing ; Takubo, Naoko ; Wang, Yihua. / In vivo myosin step-size from zebrafish skeletal muscle. In: Open Biology. 2016 ; Vol. 6, No. 5.
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