Quantitative high-precision imaging of myosin-dependent filamentous actin dynamics.
Quantitative high-precision imaging of myosin-dependent filamentous actin dynamics.
复制标题
肌球蛋白依赖性丝状肌动蛋白动力学的定量高精度成像。
DOI：
10.1007/s10974-019-09541-x
复制
发表时间：
2020
影响因子：
2.7
通讯作者：
Watanabe Naoki
中科院分区：
文献类型：
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作者：
Yamashiro Sawako;Watanabe Naoki
近几十年来，人们已经做出了相当大的努力来了解施加到肌动蛋白网络的机械应力如何改变肌动蛋白的组装和拆卸动力学。然而，关于体外和细胞中的问题，存在相互矛盾的报道。在这篇综述中，我们讨论了有关以前的定量活细胞实验，试图评估肌球蛋白调节丝状肌动蛋白（F-肌动蛋白）营业额的关注。特别是，我们强调了一个错误的产生机制，在定量活细胞成像，即对流引起的错误分布肌动蛋白结合探针。使用我们改进的基于电穿孔的单分子斑点（eSiMS）显微镜技术在单分子水平上直接观察肌动蛋白周转克服了这些问题。我们介绍了我们最近的单分子分析，明确表明肌球蛋白依赖的调节F-肌动蛋白在活细胞中的稳定性。我们还讨论了eSiMS显微镜在分析横纹肌细胞肌动蛋白重塑中的可能应用。
Over recent decades, considerable effort has been made to understand how mechanical stress applied to the actin network alters actin assembly and disassembly dynamics. However, there are conflicting reports concerning the issue both in vitro and in cells. In this review, we discuss concerns regarding previous quantitative live-cell experiments that have attempted to evaluate myosin regulation of filamentous actin (F-actin) turnover. In particular, we highlight an error-generating mechanism in quantitative live-cell imaging, namely convection-induced misdistribution of actin-binding probes. Direct observation of actin turnover at the single-molecule level using our improved electroporation-based Single-Molecule Speckle (eSiMS) microscopy technique overcomes these concerns. We introduce our recent single-molecule analysis that unambiguously demonstrates myosin-dependent regulation of F-actin stability in live cells. We also discuss the possible application of eSiMS microscopy in the analysis of actin remodeling in striated muscle cells.