Yamamoto Takenori, Tachikawa Aiko, Terauchi Satsuki, Yamashita Kikuji, Kataoka Masatoshi, Terada Hiroshi, Shinohara Yasuo
Institute for Genome Research, Faculty of Pharmaceutical Sciences, University of Tokushima, Japan.
Eur J Biochem. 2004 Sep;271(17):3573-9. doi: 10.1111/j.1432-1033.2004.04294.x.
3,3'-Dipropyl-2,2'-thiadicarbocyanine iodide [DiS-C(3)(5)], often used as a tracer dye to assess the mitochondrial membrane potential, was investigated in detail regarding its effects on the structure and function of isolated mitochondria. As reported previously, DiS-C(3)(5) had an inhibitory effect on NADH-driven mitochondrial electron transfer. On the contrary, in the presence of inorganic phosphate, DiS-C(3)(5) showed dose-dependent biphasic effects on mitochondria energized by succinate. At higher concentrations, such as 50 micro m, DiS-C(3)(5) accelerated mitochondrial oxygen consumption. Measurements of the permeability of DiS-C(3)(5)-treated mitochondrial membranes to poly(ethylene glycol) and analysis of mitochondrial configuration by transmission electron microscopy revealed that the accelerating effect of DiS-C(3)(5) on mitochondrial oxygen consumption reflects the induction of the mitochondrial permeability transition (PT). When the mitochondrial PT was induced by DiS-C(3)(5), release of mitochondrial cytochrome c was observed, as in the case of the PT induced by Ca(2+). On the contrary, at a low concentration such as 5 micro m, DiS-C(3)(5) showed an inhibitory effect on the latent oxygen consumption by mitochondria. This effect was shown to reflect inhibition of the PT induced by a low concentration of Ca(2+). Furthermore, in the absence of inorganic phosphate, DiS-C(3)(5) caused mitochondrial swelling. Under this condition, DiS-C(3)(5) caused changes in the membrane status of the mitochondria, but did not induce a release of mitochondrial cytochrome c.
3,3'-二丙基-2,2'-硫代二碳菁碘化物[DiS-C(3)(5)]常被用作示踪染料来评估线粒体膜电位，本文详细研究了其对分离线粒体结构和功能的影响。如先前报道，DiS-C(3)(5)对NADH驱动的线粒体电子传递有抑制作用。相反，在无机磷酸盐存在的情况下，DiS-C(3)(5)对由琥珀酸激发的线粒体呈现剂量依赖性的双相效应。在较高浓度下，如50微摩尔，DiS-C(3)(5)会加速线粒体的氧气消耗。对经DiS-C(3)(5)处理的线粒体膜对聚乙二醇的通透性测量以及通过透射电子显微镜对线粒体形态的分析表明，DiS-C(3)(5)对线粒体氧气消耗的加速作用反映了线粒体通透性转换(PT)的诱导。当DiS-C(3)(5)诱导线粒体PT时，会观察到线粒体细胞色素c的释放，这与Ca(2+)诱导的PT情况相同。相反，在低浓度如5微摩尔时，DiS-C(3)(5)对线粒体潜在的氧气消耗有抑制作用。这种作用被证明反映了对低浓度Ca(2+)诱导的PT的抑制。此外，在没有无机磷酸盐的情况下，DiS-C(3)(5)会导致线粒体肿胀。在这种情况下，DiS-C(3)(5)会引起线粒体膜状态的变化，但不会诱导线粒体细胞色素c的释放。
