成像心肌细胞中的钙火花。
Imaging calcium sparks in cardiac myocytes.
作者信息
Guatimosim Silvia, Guatimosim Cristina, Song Long-Sheng
机构信息
Department of Physiology and Biophysics, ICB, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil.
出版信息
Methods Mol Biol. 2011;689:205-14. doi: 10.1007/978-1-60761-950-5_12.
Calcium ions play fundamental roles in many cellular processes in virtually all type of cells. The use of Ca(2+) sensitive fluorescent indicators has proven to be an indispensable tool for studying the spatio-temporal dynamics of intracellular calcium (Ca(2+)). With the aid of laser scanning confocal microscopy and new generation of Ca(2+) indicators, highly localized, short-lived Ca(2+) signals, namely Ca(2+) sparks, were revealed as elementary Ca(2+) release events during excitation-contraction coupling in cardiomyocytes. Since the discovery of Ca(2+) sparks in 1993, the demonstration of dynamic Ca(2+) micro-domains in living cardiomyocytes has revolutionized our understanding of Ca(2+)-mediated signal transduction in normal and diseased hearts. In this chapter, we have described a commonly used method for recording local and global Ca(2+) signals in cardiomyocytes using the fluorescent indicator fluo-4 acetoxymethyl (AM) and laser scanning confocal microscopy.
钙离子在几乎所有类型的细胞的许多细胞过程中都发挥着重要作用。事实证明，使用对Ca(2+)敏感的荧光指示剂是研究细胞内钙（[Ca(2+)]i）时空动态的不可或缺的工具。借助激光扫描共聚焦显微镜和新一代Ca(2+)指示剂，高度局部化、短暂的Ca(2+)信号，即Ca(2+)火花，被揭示为心肌细胞兴奋-收缩偶联过程中的基本Ca(2+)释放事件。自1993年发现Ca(2+)火花以来，活心肌细胞中动态Ca(2+)微区的证明彻底改变了我们对正常和患病心脏中Ca(2+)介导的信号转导的理解。在本章中，我们描述了一种使用荧光指示剂fluo-4乙酰氧基甲基（AM）和激光扫描共聚焦显微镜记录心肌细胞局部和全局Ca(2+)信号的常用方法。
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