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哺乳动物骨骼肌中融合事件的检测。

Detection of fusion events in Mammalian skeletal muscle.

作者信息

Suhr Frank

机构信息

Department of Molecular and Cellular Sport Medicine, Institute of Cardiovascular Research and Sport Medicine, German Sport University Cologne, Am Sportpark Mungersdorf 6, Cologne, 50933, Germany,

出版信息

Methods Mol Biol. 2015;1313:115-29. doi: 10.1007/978-1-4939-2703-6_8.

Abstract

Cell fusion events are essential for the maintenance of skeletal muscle tissue and during its repair processes after damage. However, these mechanisms have not come much into focus in the recent years. Different methods can be used to assess ongoing cell fusion events in adult skeletal muscle tissue. Among these methods, confocal microscopy, western blotting, and quantitative polymerase chain reactions are ideal, since they provide concerted information about cell fusion events going on in skeletal muscle tissue at both qualitative and quantitative levels. Confocal microscopy allows for the visualization of exact localizations of cell fusion events in adult skeletal muscle. Western blotting allows for a semiquantitative evaluation of protein levels involved and associated with cell fusions events. Finally, quantitative polymerase chain reaction is a valuable tool to precisely assess mRNA levels of genes involved and associated with cell fusions events. In addition to the investigation if cell fusion markers in skeletal muscle tissue, in vitro cell culture systems (e.g., C2C12 cells) can be used to study cell fusions events in a highly standardized system in order to obtain detailed information about genes and proteins involved in these processes. Here, confocal microscopy, western blotting, and quantitative polymerase chain reaction are described as methods to investigate cell fusion events and how a C2C12 cell culture system can be run to support the studies of adult muscle tissue.

摘要

细胞融合事件对于维持骨骼肌组织以及在其损伤后的修复过程中至关重要。然而,近年来这些机制并未受到太多关注。可以使用不同的方法来评估成年骨骼肌组织中正在进行的细胞融合事件。在这些方法中,共聚焦显微镜、蛋白质免疫印迹和定量聚合酶链反应是理想的,因为它们在定性和定量水平上都能提供有关骨骼肌组织中正在发生的细胞融合事件的协同信息。共聚焦显微镜能够可视化成年骨骼肌中细胞融合事件的确切定位。蛋白质免疫印迹允许对参与细胞融合事件的蛋白质水平进行半定量评估。最后,定量聚合酶链反应是精确评估参与细胞融合事件的基因的mRNA水平的有价值工具。除了研究骨骼肌组织中的细胞融合标志物外,体外细胞培养系统(如C2C12细胞)可用于在高度标准化的系统中研究细胞融合事件,以便获得有关这些过程中涉及的基因和蛋白质的详细信息。在此,共聚焦显微镜、蛋白质免疫印迹和定量聚合酶链反应被描述为研究细胞融合事件的方法,以及如何运行C2C12细胞培养系统以支持对成年肌肉组织的研究。

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