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离体心肌细胞兴奋-收缩偶联过程中线粒体钙瞬变的物种依赖性

Species dependence of mitochondrial calcium transients during excitation-contraction coupling in isolated cardiomyocytes.

作者信息

Griffiths E J

机构信息

Bristol Heart Institute, University of Bristol, United Kingdom.

出版信息

Biochem Biophys Res Commun. 1999 Sep 24;263(2):554-9. doi: 10.1006/bbrc.1999.1311.

Abstract

Whether mitochondrial Ca(2+) transport is rapid enough to respond to changes in cytosolic [Ca(2+)] (Ca(2+)) which occur during excitation-contraction coupling in the heart is controversial; different results wereobtained with different techniques and different species. In this study mitochondrial [Ca(2+)] (Ca(2+)) was measured in indo-1/AM-loaded myocytes from rat and guinea-pig hearts where the cytosolic indo-1 had been removed by extended incubation of cells at 37 degrees C ("heat treatment"). The mitochondrial origin of the remaining fluorescence was confirmed by sensitivity of the indo-1 signal to ruthenium red. In resting rat myocytes, Ca(2+) was lower than Ca(2+), whereas in guinea-pig cells Ca(2+) was higher than Ca(2+). Upon electrical stimulation of cells, no change occurred in Ca(2+) in rat myocytes. However, in guinea-pig cells mitochondrial Ca(2+) transients were clearly visible with a mean indo-1 ratio amplitude of 0.153 +/- 0.2 (n = 20), compared with 0.306 +/- 0.02 (n = 25), p < 0.001, prior to heat treatment. These observations suggest significant differences in mitochondrial Ca(2+) transport in cardiomyocytes from different species.

摘要

线粒体Ca(2+)转运是否足够迅速,以响应心脏兴奋-收缩偶联过程中发生的胞质[Ca(2+)]([Ca(2+)]c)变化,这一点存在争议;使用不同技术和不同物种获得了不同结果。在本研究中,在来自大鼠和豚鼠心脏的indo-1/AM负载的心肌细胞中测量线粒体[Ca(2+)]([Ca(2+)]m),其中通过在37℃下长时间孵育细胞(“热处理”)去除了胞质indo-1。通过indo-1信号对钌红的敏感性证实了剩余荧光的线粒体来源。在静息大鼠心肌细胞中,[Ca(2+)]m低于[Ca(2+)]c,而在豚鼠细胞中[Ca(2+)]m高于[Ca(2+)]c。对细胞进行电刺激后,大鼠心肌细胞中的[Ca(2+)]m没有变化。然而,在豚鼠细胞中,线粒体Ca(2+)瞬变清晰可见,平均indo-1比率幅度为0.153±0.2(n = 20),而热处理前为0.306±0.02(n = 25),p < 0.001。这些观察结果表明不同物种心肌细胞中线粒体Ca(2+)转运存在显著差异。

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