A CARBOCYANINE DYE, DIOC6(3), ACTS AS A MITOCHONDRIAL PROBE IN HUMAN-NEUTROPHILS
A CARBOCYANINE DYE, DIOC6(3), ACTS AS A MITOCHONDRIAL PROBE IN HUMAN-NEUTROPHILS
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DOI：
10.1016/s0006-291x(82)80076-4
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发表时间：
1982-01-01
影响因子：
3.1
通讯作者：
WEISSMANN, G
中科院分区：
文献类型：
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作者：
KORCHAK, HM;RICH, AM;WEISSMANN, G
使用羰花青染料DiOC 6（3）（3，3“-二己基氧羰花青碘化物）监测人嗜中性粒细胞的膜电位变化。荧光显微镜显示，碳菁染料DiOC 6（3）主要分配到亚细胞器，可能是线粒体。荧光光谱表明，激活的中性粒细胞的f-Met-Leu-Phe和佛波醇肉豆蔻酸酯乙酸酯是由一个短暂的滞后期，其中没有观察到荧光的变化，然后由一个快速的荧光损失，一个明显的去极化。荧光显微镜显示，这种荧光损失来自线粒体。电子传递抑制剂氰化物和鱼藤酮和氧化磷酸化解偶联剂CCCP [羰基氰化物-间氯苯腙]和二硝基苯酚抑制荧光损失。因此，DiOC 6（3）主要报告线粒体膜电位的变化，而不是跨质膜膜电位的变化。使用碳菁染料和亲脂性阳离子TPMP+ [三苯基甲基磷离子]获得的膜电位结果的明显不一致性可以解释为探针分布差异的基础上。
Membrane potential changes were monitored in human neutrophils using the carbocyanine dye DiOC6(3) (3,3''-dihexyloxacarbocyanine iodide). Fluorescence microscopy demonstrated that the carbocyanine dye DiOC6(3) partitions predominantly into subcellular organelles, probably mitochondria. Fluorescence spectroscopy demonstrated that activation of the neutrophil by f-Met-Leu-Phe and phorbol myristate acetate is followed by a short lag period where no change in fluorescence was observed, followed by a rapid loss of fluorescence, an apparent depolarization. Fluorescence microscopy demonstrated that this loss of fluorescence was from the mitochondria. The fluorescence loss was inhibited by the electron transport inhibitors cyanide and rotenone and by the oxidative phosphorylation uncouplers CCCP [carbonyl cyanide-m-chlorophenylhydrazone] and dinitrophenol. Thus DiOC6(3) predominantly reported changes in mitochondrial membrane potential and not transplasmalemmal membrane potential. The apparent discordance in membrane potential results obtained using carbocyanine dye and the lipophilic cation TPMP+ [triphenylmethyl phosphonium ion] may be explained on the basis of differences in probe distribution.