Tashiro M, Konishi M
Department of Physiology, The Jikei University School of Medicine, Tokyo 105 - 8461, Japan.
Am J Physiol Cell Physiol. 2000 Dec;279(6):C1955-62. doi: 10.1152/ajpcell.2000.279.6.C1955.
Cytoplasmic concentration of Mg(2+) (Mg(2+)) was measured with a fluorescent indicator furaptra in ventricular myocytes enzymatically dissociated from rat hearts (25 degrees C). To study Mg(2+) transport across the cell membrane, cells were treated with ionomycin in Ca(2+)-free (0.1 mM EGTA) and high-Mg(2+) (10 mM) conditions to facilitate passive Mg(2+) influx. Rate of rise of Mg(2+) due to the net Mg(2+) influx was significantly smaller in the presence of 130 mM extracellular Na(+) than in its absence. We also tested the extracellular Na(+) dependence of the net Mg(2+) efflux from cells loaded with Mg(2+). After Mg(2+) was raised by ionomycin and high Mg(2+) to the level 0.5-0.6 mM above the basal value ( approximately 0.7 mM), washout of ionomycin and lowering extracellular [Mg(2+)] to 1.2 mM caused rapid decline of Mg(2+) in the presence of 140 mM Na(+). This net efflux of Mg(2+) was completely inhibited by withdrawal of extracellular Na(+) and was largely attenuated by imipramine, a known inhibitor of Na(+)/Mg(2+) exchange, with 50% inhibition at 79 microM. The relation between the rate of net Mg(2+) efflux and extracellular Na(+) concentration (Na(+)) had a Hill coefficient of 2 and Na(+) at half-maximal rate of 82 mM. These results demonstrate the presence of Na(+) gradient-dependent Mg(2+) transport, which is consistent with Na(+)/Mg(2+) exchange, in cardiac myocytes.
在25摄氏度下,用荧光指示剂呋喃普特拉测定从大鼠心脏酶解分离的心室肌细胞中镁离子(Mg(2+))的胞浆浓度([Mg(2+)]i)。为了研究Mg(2+)跨细胞膜的转运,细胞在无钙(0.1 mM乙二醇双四乙酸)和高镁(10 mM)条件下用离子霉素处理,以促进Mg(2+)的被动内流。在存在130 mM细胞外钠离子(Na(+))的情况下,由于净Mg(2+)内流导致的[Mg(2+)]i上升速率明显小于不存在时。我们还测试了细胞外Na(+)对加载了Mg(2+)的细胞净Mg(2+)外流的依赖性。在用离子霉素和高镁将[Mg(2+)]i提高到比基础值(约0.7 mM)高0.5 - 0.6 mM的水平后,洗脱离子霉素并将细胞外[Mg(2+)]降低到1.2 mM,在存在140 mM Na(+)的情况下,[Mg(2+)]i迅速下降。这种Mg(2+)的净外流在去除细胞外Na(+)时完全被抑制,并且在很大程度上被已知的Na(+)/Mg(2+)交换抑制剂丙咪嗪减弱,在79 microM时抑制率为50%。净Mg(2+)外流速率与细胞外Na(+)浓度([Na(+)]o)之间的关系具有2的希尔系数,半最大速率时的[Na(+)]o为82 mM。这些结果表明心肌细胞中存在与Na(+)梯度相关的Mg(2+)转运,这与Na(+)/Mg(2+)交换一致。