Revealing t-tubules in striated muscle with new optical super-resolution microscopy techniques

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Isuru D. Jayasinghe
Alexander H. Clowsley
Michelle Munro
Yufeng Hou
David J. Crossman
Christian Soeller *
(*) Corresponding Author:
Christian Soeller | c.soeller@exeter.ac.uk

Abstract

The t-tubular system plays a central role in the synchronisation of calcium signalling and excitation-contraction coupling in most striated muscle cells. Light microscopy has been used for imaging t-tubules for well over 100 years and together with electron microscopy (EM), has revealed the three-dimensional complexities of the t-system topology within cardiomyocytes and skeletal muscle fibres from a range of species. The emerging super-resolution single molecule localisation microscopy (SMLM) techniques are offering a near 10-fold improvement over the resolution of conventional fluorescence light microscopy methods, with the ability to spectrally resolve nanometre scale distributions of multiple molecular targets. In conjunction with the next generation of electron microscopy, SMLM has allowed the visualisation and quantification of intricate t-tubule morphologies within large areas of muscle cells at an unprecedented level of detail. In this paper, we review recent advancements in the t-tubule structural biology with the utility of various microscopy techniques. We outline the technical considerations in adapting SMLM to study t-tubules and its potential to further our understanding of the molecular processes that underlie the sub-micron scale structural alterations observed in a range of muscle pathologies.

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Author Biographies

Isuru D. Jayasinghe, Biomedical Physics, University of Exeter

Associate research fellow, College of Physics, University of Exeter

Alexander H. Clowsley, Biomedical Physics, University of Exeter

PhD candidate, College of Physics, University of Exeter

Michelle Munro, Department of Physiology, The University of Auckland

PhD candidate, Dept of Physiology

Yufeng Hou, Department of Physiology, The University of Auckland

PhD candidate, Dept of Physiology

David J. Crossman, Department of Physiology, The University of Auckland

Research Fellow, Dept of Physiology

Christian Soeller, Biomedical Physics, University of Exeter, United Kingdom; Department of Physiology, The University of Auckland

Chair in Physical Cell Biology,
Biomedical Physics