Multiple effects of DiS-C3(5) on mitochondrial structure and function.
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.
Institute for Genome Research, Faculty of Pharmaceutical Sciences, University of Tokushima, Japan.
Takenori
T
Yamamoto
Aiko
A
Tachikawa
Satsuki
S
Terauchi
Kikuji
K
Yamashita
Masatoshi
M
Kataoka
Hiroshi
H
Terada
Yasuo
Y
Shinohara
D000818:Animals; D052160:Benzothiazoles; D002118:Calcium / Q000378:metabolism; D002232:Carbocyanines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004579:Electron Transport / Q000187:drug effects / Q000502:physiology; D005456:Fluorescent Dyes / Q000378:metabolism*; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism* / Q000648:ultrastructure; D015394:Molecular Structure; D009243:NAD / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010539:Permeability; D010710:Phosphates / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D052160:Benzothiazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D010710:Phosphates; D009243:NAD; C012944:3,3'-dipropyl-2,2'-thiadicarbocyanine; D045304:Cytochromes c; D010100:Oxygen; D002118:Calcium