Tashiro M, Konishi M, Iwamoto T, Shigekawa M, Kurihara S
Department of Physiology, The Jikei University School of Medicine, Tokyo, Japan.
Pflugers Arch. 2000 Oct;440(6):819-27. doi: 10.1007/s004240000384.
Cytoplasmic concentrations of Ca2+ ([Ca2+]i) and Mg2+ ([Mg2+]i) were measured with fluorescent indicators in CCL39 cells, a cell line established from Chinese hamster lung fibroblasts, transfected with complementary deoxyribonucleic acid (cDNA) of the Na+-Ca2+ exchanger isolated either from canine heart (NCX1) or from rat brain (NCX3). Raising extracellular [Mg2+] to 10 mM increased Mg2+ influx and the resultant change in [Mg2+]i (delta[Mg2+]i) was monitored with furaptra under Ca2+-free conditions. In control (vector-transfected) cells, delta[Mg2+]i at 45 min was similar with or without extracellular Na+ (130 mM or 0 mM) and when [Na+]i was raised by 1 mM ouabain treatment. delta[Mg2+]i in NCX1-transfected cells was attenuated significantly in the presence of 130 mM Na+, but became comparable to (or slightly larger than) that in control cells on either removal of extracellular Na+ or treatment with 1 mM ouabain. Cells expressing NCX3 showed an intermediate dependence of delta[Mg2+]i on Na+, probably reflecting a lower degree of expression of the exchanger protein. Extracellular Na+-dependent changes in [Ca2+]i (measured with fura-2 in the presence of extracellular Ca2+ and 10 microM ionomycin, a Ca2+ ionophore) were minimal in control cells, marked in the NCX1-transfected cells and intermediate in the NCX3-transfected cells. These results suggest that the Na+-Ca2+ exchanger (either NCX1 or NCX3) can transport Mg2+ and may play a role in the extrusion of magnesium from cells.
用荧光指示剂测量了CCL39细胞(一种源自中国仓鼠肺成纤维细胞的细胞系)胞质中Ca2+([Ca2+]i)和Mg2+([Mg2+]i)的浓度,该细胞系转染了从犬心脏分离的Na+-Ca2+交换体(NCX1)或大鼠脑分离的Na+-Ca2+交换体(NCX3)的互补脱氧核糖核酸(cDNA)。将细胞外[Mg2+]提高到10 mM会增加Mg2+内流,并在无Ca2+条件下用氟罗帕特拉监测由此产生的[Mg2+]i变化(δ[Mg2+]i)。在对照(载体转染)细胞中,45分钟时的δ[Mg2+]i在有或无细胞外Na+(130 mM或0 mM)以及用1 mM哇巴因处理使[Na+]i升高时相似。在130 mM Na+存在下,NCX1转染细胞中的δ[Mg2+]i显著减弱,但在去除细胞外Na+或用1 mM哇巴因处理后变得与对照细胞相当(或略大于对照细胞)。表达NCX3的细胞显示δ[Mg2+]i对Na+的依赖性处于中间水平,这可能反映了交换体蛋白较低的表达程度。在细胞外Ca2+存在且有10 μM离子霉素(一种Ca2+离子载体)的情况下用fura-2测量的细胞外Na+依赖性[Ca2+]i变化在对照细胞中最小,在NCX1转染细胞中明显,在NCX3转染细胞中处于中间水平。这些结果表明,Na+-Ca2+交换体(NCX1或NCX3)可以转运Mg2+,并可能在细胞内镁的外排中起作用。