Jayasinghe Isuru D, Clowsley Alexander H, Munro Michelle, Hou Yufeng, Crossman David J, Soeller Christian
Biomedical Physics, University of Exeter , UK, New Zealand.
Department of Physiology, The University of Auckland , New Zealand.
Eur J Transl Myol. 2014 Dec 24;25(1):4747. doi: 10.4081/ejtm.2015.4747. eCollection 2015 Jan 7.
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.
在大多数横纹肌细胞中，横管系统在钙信号同步以及兴奋 - 收缩偶联过程中发挥着核心作用。光学显微镜用于横管成像已有100多年历史，并且与电子显微镜（EM）一起，揭示了一系列物种的心肌细胞和骨骼肌纤维内横管系统拓扑结构的三维复杂性。新兴的超分辨率单分子定位显微镜（SMLM）技术相比传统荧光光学显微镜方法的分辨率提高了近10倍，能够在光谱上分辨多个分子靶点的纳米级分布。结合下一代电子显微镜，SMLM使人们能够以前所未有的详细程度可视化和量化肌肉细胞大面积内复杂的横管形态。在本文中，我们回顾了利用各种显微镜技术在横管结构生物学方面的最新进展。我们概述了将SMLM应用于研究横管的技术考量，以及它在进一步加深我们对一系列肌肉疾病中观察到的亚微米级结构改变所涉及分子过程理解方面的潜力。
