Lee Seungah, Lee Hee Gu, Kang Seong Ho
Department of Chemistry and Research Institute of Physics and Chemistry (RINPAC), Chonbuk National University, Jeonju 561-756, South Korea.
Anal Chem. 2009 Jan 15;81(2):538-42. doi: 10.1021/ac8013324.
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.
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。