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用离子选择性微电极测量青蛙骨骼肌纤维中的细胞内游离镁。

Intracellular free magnesium in frog skeletal muscle fibres measured with ion-selective micro-electrodes.

作者信息

Alvarez-Leefmans F J, Gamiño S M, Giraldez F, González-Serratos H

出版信息

J Physiol. 1986 Sep;378:461-83. doi: 10.1113/jphysiol.1986.sp016230.

Abstract

Intracellular free Mg2+ concentration [( Mg2+]i) was measured in frog skeletal muscle fibres, using Mg2+-selective micro-electrodes based on the neutral ligand ETH-1117. In calibration solutions the electrodes showed significant interference from K+, and to a lesser extent from Na+, at concentrations found intracellularly. Therefore, in order to calibrate the electrodes properly, it was necessary first to obtain an accurate value for intracellular free Na+ and K+ concentrations ([Na+]i and [K+]i), using the appropriate liquid ion exchanger micro-electrodes. In fibres from muscles maintained in Ringer solution, the mean value for [Na+]i was 6.2 +/- 0.4 mM (S.E. of mean; n = 20 fibres in five muscles), while [K+]i was 104 +/- 1.7 mM (range 83-122 mM; n = 25 fibres in eight muscles). Due to the substantial variability found for [K+]i, not only between fibres from different muscles, but also between fibres belonging to the same muscle, it was necessary to measure [Mg2+]i and [K+]i simultaneously in the same fibre to determine as accurately as possible the degree of K+ interference on Mg2+-selective micro-electrode response. In nineteen fibres from six muscles maintained in Ringer solution, the mean [K+]i was 91.7 +/- 2.7 mM (range 71-110 mM), while the mean [Mg2+]i was 0.80 +/- 0.07 mM (range 0.2-1.2 mM). The mean resting potential was -79.3 +/- 0.4 mV (S.E. of mean). In fifteen fibres from four muscles equilibrated in Ringer solution containing 0.5 mM-Mg2+, the mean [K+]i was 115.5 +/- 0.1 mM (range 97-129 mM) and the mean [Mg2+]i measured simultaneously in the same fibres was 1.69 +/- 0.21 mM (range 0.2-2.7 mM). The mean resting potential was -83 +/- 0.7 mV. The mean [K+]i and [Mg2+]i found in these fibres was significantly higher (P less than 0.0001) than those measured in fibres from muscles maintained in standard Ringer solution (i.e. without external Mg2+). Possible explanations for this finding are discussed. Whether in the presence (0.5 mM) or in the absence of external Mg2+, our values for [Mg2+]i are distinctly lower than those previously reported by others, using the same type of Mg2+-selective micro-electrodes but calibrated simply from assumptions about the actual level of K+ and Na+ interference on Mg2+-selective micro-electrode response.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

利用基于中性配体ETH - 1117的镁离子选择性微电极,测量了青蛙骨骼肌纤维中的细胞内游离镁离子浓度[Mg2+]i。在校准溶液中,电极在细胞内发现的钾离子浓度下显示出显著的钾离子干扰,钠离子干扰程度较小。因此,为了正确校准电极,首先需要使用合适的液体离子交换微电极获得细胞内游离钠离子和钾离子浓度([Na+]i和[K+]i)的准确值。在置于林格溶液中的肌肉纤维中,[Na+]i的平均值为6.2±0.4 mM(平均值的标准误;来自五块肌肉的20根纤维),而[K+]i为104±1.7 mM(范围83 - 122 mM;来自八块肌肉的25根纤维)。由于[K+]i不仅在不同肌肉的纤维之间,而且在同一肌肉的纤维之间存在很大差异,因此有必要在同一根纤维中同时测量[Mg2+]i和[K+]i,以尽可能准确地确定钾离子对镁离子选择性微电极响应的干扰程度。在置于林格溶液中的六块肌肉的19根纤维中,[K+]i的平均值为91.7±2.7 mM(范围71 - 110 mM),而[Mg2+]i的平均值为0.80±0.07 mM(范围0.2 - 1.2 mM)。平均静息电位为 - 79.3±0.4 mV(平均值的标准误)。在置于含0.5 mM - Mg2+的林格溶液中平衡的四块肌肉的15根纤维中,[K+]i的平均值为115.5±0.1 mM(范围97 - 129 mM),并且在同一纤维中同时测量的[Mg2+]i的平均值为1.69±0.21 mM(范围0.2 - 2.7 mM)。平均静息电位为 - 83±0.7 mV。在这些纤维中发现的[K+]i和[Mg2+]i的平均值显著高于(P小于0.0001)在置于标准林格溶液(即无外部Mg2+)中的肌肉纤维中测量的值。讨论了这一发现的可能解释。无论是否存在(0.5 mM)外部Mg2+,我们的[Mg2+]i值明显低于其他人先前报道的值,他们使用相同类型的镁离子选择性微电极,但仅根据关于钾离子和钠离子对镁离子选择性微电极响应的实际干扰水平的假设进行校准。(摘要截断于400字)

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