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对单个活的心室肌细胞内Mg2+信号传导和波的实时观察。

Real-time observations of intracellular Mg2+ signaling and waves in a single living ventricular myocyte cell.

作者信息

Lee Seungah, Lee Hee Gu, Kang Seong Ho

机构信息

Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University, Jeonju 561-756, South Korea.

出版信息

Anal Chem. 2009 Jan 15;81(2):538-42. doi: 10.1021/ac8013324.

Abstract

Despite the important regulatory role of Mg(2+) in metabolic pathways, its underlying mechanism is not completely understood at the single-cell level. This study examined the propagation and dynamics of Mg(2+) signaling across the cell membrane by employing the real-time visualization of intracellular Mg(2+) waves in living ventricular myocytes using a combination of total internal reflection fluorescence microscopy and Nomarski differential interference contrast. Real-time Mg(2+) waves and sparks in a living cell membrane were observed using a fluorescent Mg(2+) indicator (mag-fluo-4-AM) in the concentration range of 5 aM-5 muM. The intracellular locations of the fluorescent Mg(2+) indicator were confirmed by adding Na(+)ATP. The Mg(2+) sparks and waves showed random temporal propagation patterns in nonhomogeneous substructures. These results show that spatiotemporal intracellular Mg(2+) signaling information can be obtained for individual living cells.

摘要

尽管镁离子(Mg(2+))在代谢途径中具有重要的调节作用,但其在单细胞水平的潜在机制尚未完全阐明。本研究通过结合全内反射荧光显微镜和诺马斯基微分干涉对比技术,对活的心室肌细胞内镁离子波进行实时可视化,从而研究镁离子信号在细胞膜上的传播和动态变化。使用浓度范围为5 aM至5 μM的荧光镁离子指示剂(mag-fluo-4-AM),观察到活细胞膜上的实时镁离子波和火花。通过添加Na(+)ATP来确认荧光镁离子指示剂在细胞内的位置。镁离子火花和波在非均匀亚结构中呈现出随机的时间传播模式。这些结果表明,可以获取单个活细胞的时空细胞内镁离子信号信息。

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