Specific requirement for inorganic phosphate for induction of bilayer membrane conductance by the cationic uncoupler carbocyanine dye

Terada, Hiroshi; Nagamune, Hideaki; Osaki, Yumi; Yoshikawa, Kenichi

doi:10.1016/0005-2736(81)90320-5

Cited by 17 publications

(9 citation statements)

References 12 publications

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“…Then, they were energized by the addition of either 10 mM succinate (plus 0.5 lgAEmg )1 protein rotenone) or 10 mM glutamate and 10 mM malate as respiratory substrates. Rates of mitochondrial oxygen consumption at 25°C were measured by use of a Clark oxygen electrode (YSI 5331;Yellow Springs Instrument Co., Yellow Springs, OH, USA) 6 . When the inhibitory effects of DiS-C 3 (5) on the mitochondrial oxygen consumption were evaluated, the protonophoric uncoupler 3,5-di-tert-butyl-4-hydroxy-benzylidene malononitrile (SF6847) was utilized to induce maximum oxygen consumption.…”

Section: Measurement Of Mitochondrial Oxygen Consumption and Swellingmentioning

confidence: 99%

Multiple effects of DiS‐C₃(5) on mitochondrial structure and function

Yamamoto¹,

Tachikawa²,

Terauchi³

et al. 2004

European Journal of Biochemistry

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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 lM, 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 lM, 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.Keywords: cyanine dye; cytochrome c; DiS-C 3 (5); mitochondria; permeability transition.The mitochondrial inner membrane is highly impermeable even to small solutes and ions. However, under certain conditions, such as in the presence of Ca 2+ and inorganic phosphate (P i ), the inner mitochondrial membrane becomes permeable to solutes and ions up to 1500 Da. This phenomenon is referred to as the mitochondrial permeability transition (PT), and PT is believed to reflect the opening of a proteinaceous pore [1][2][3].In the field of biochemistry, cyanine dyes are often employed as an indicator dye to assess the mitochondrial membrane potential [4,5]. In our previous studies, we characterized the effects of cyanine dyes such as 2,2¢-{3-[2-(3-butyl-4-methyl-2-thiazolin-2-ylidene)ethylidene]propenylene}-bis(3-butyl-4-methyl thiazolinium iodide) [TriS-C 4 (5)] and 2,2¢-{3-[2-(3-heptyl-4-methyl-2-thiazolin-2-ylidene) ethylidene] propenylene}-bis(3-heptyl-4-methyl thiazolinium iodide) [TriS-C 7 (5)], both of which have three heterocylic groups, on mitochondrial structure and function. These cyanine dyes accelerated mitochondrial oxygen consumption only in the presence of P i in the incubation medium [6][7][8]. Furthermore, the accelerating effects of these cyanine dyes on the mitochondrial oxygen consumption were attributable mainly to the induction of the mitochondrial PT [9]. However, different from the classical PT induced by Ca , that induced by these cyanine dyes was o...

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Section: Measurement Of Mitochondrial Oxygen Consumption and Swellingmentioning

confidence: 99%

Multiple effects of DiS‐C₃(5) on mitochondrial structure and function

Yamamoto¹,

Tachikawa²,

Terauchi³

et al. 2004

European Journal of Biochemistry

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“…[3][4][5] Mitochondrial respiration was measured with a Clark oxygen electrode (Yellow Springs, model 5331) at 25°C. For this, mitochondria were first suspended in +Pi medium (200 mM sucrose and 10 mM potassium phosphate buffer, pH 7.4) or -Pi medium (200 mM sucrose, 10 mM KCl, and 10 mM Tris-Cl buffer; pH 7.4) to make a final concentration of 0.7 mg protein/ml.…”

Section: Isolation Of Mitochondria and Measurement Ofmentioning

confidence: 99%

“…Furthermore, we also examined the effect of another cyanine dye, tri-S-C 7 (5), having a saturated hydrocarbon side chain (n = 7) longer than that of tri-S-C 4 (5) (n = 4), because we reported earlier that tri-S-C 7 (5) showed similar actions toward mitochondrial functions as tri-S-C 4 (5). 3,4) For a control, the effect of CsA on PT induced by Ca 2+ was also examined. As reported previously, in the presence of Pi, cyanine dyes tri-S-C 4 (5) and tri-S-C 7 (5) caused remarkable acceleration of mitochondrial respiration that was accompanied by mitochondrial swelling.…”

Section: In the Presence Of Pi Cyanine Dyes Accelerated Mitochondriamentioning

confidence: 99%

“…1,2) However, at higher concentrations than those used for monitoring the membrane potential, some of them were found to act as uncouplers of mitochondrial oxidative phosphorylation. [3][4][5][6] Unlike the weakly acidic uncouplers such as FCCP and SF6847, [7][8][9][10] the uncoupling action of cyanine dyes requires inorganic phosphate (Pi) and is accompanied by mitochondrial swelling. These features are very similar to those induced by Ca 2+ .…”

Section: Introductionmentioning

confidence: 99%

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Three-Way Effect of Cyanine Dye on the Structure and Function of Mitochondria

Yamashita

Ichikawa

Yamamoto

et al. 2003

JOURNAL OF HEALTH SCIENCE

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Previously, we found that the cationic cyanine dye tri-S-C 4 (5) uncoupled mitochondrial oxidative phosphorylation by acting as an inducer of the mitochondrial permeability transition (PT). In the present study, the actions of cyanine dyes such as tri-S-C 4 (5) and tri-S-C 7 (5) on the mitochondrial structures and functions were further characterized. In the presence of inorganic phosphate (Pi), cyanine dyes were found to accelerate mitochondrial oxygen consumption that was partially sensitive to the PT inhibitor cyclosporin A (CsA). However, not only the CsAsensitive but also CsA-insensitive acceleration of mitochondrial respiration was induced by cyanine dyes; and both of them were found to be associated with the release of mitochondrial cytochrome c. On the contrary, in the absence of Pi, moderate acceleration of respiration was induced by cyanine dyes, but this respiratory effect was not associated with the induction of swelling or the release of mitochondrial cytochrome c. Thus, cyanine dyes were concluded to have 3 different effects on the mitochondrial functions depending on the Pi status.

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“…But they affect biomembrane functions by acting as hydrophobic cations (7,12,13,16,18). Recently, we found that some cyanine dyes (triS-C4 (5) and triS-C7 (5) ) are unique uncouplers of oxidative phosphorylation in mitochondria (20)(21)(22)24).…”

Section: Introductionmentioning

confidence: 99%

The cyanine dye triS-C4(5) as a cationic uncoupler of oxidative phosphorylation: Interaction with mitochondria detected by derivative spectrophotometry.

Terada

Nagamune

Morikawa

et al. 1983

Cell Struct. Funct.

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ABSTRACT.Derivative spectrophotometry was used to study the interaction of the cationic uncoupler triS-C4 (5) with mitochondria.The uncoupling action of this dye is dependent on the presence of Pi in the incubation medium.The second derivative spectrum of the dye changed with the incubation period, becoming similar to the spectrum in chloroform; but, after a time, the spectral pattern reverted to the original spectrum. The change in the spectrum in the presence of Pi was much more rapid than in its absence. The degree of spectral change agreed with the relative amount of bound dye determined directly. Thus, the spectral change reflects the binding of dye to the mitochondria, dependent on their energy state. The greater binding without Pi does not cause uncoupling but does cause shrinkage. In contrast, the lesser binding in the presence of Pi causes uncoupling and the swelling of mitochondria.These facts indicate that the dye does not penetrate the mitochondrial membrane. This refutes the idea that uncoupling by lipophilic cations is caused by the electrophoretic transfer of the uncoupler to the mitochondrial matrix space.

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Specific requirement for inorganic phosphate for induction of bilayer membrane conductance by the cationic uncoupler carbocyanine dye

Cited by 17 publications

References 12 publications

Multiple effects of DiS‐C₃(5) on mitochondrial structure and function

Multiple effects of DiS‐C₃(5) on mitochondrial structure and function

Three-Way Effect of Cyanine Dye on the Structure and Function of Mitochondria

The cyanine dye triS-C4(5) as a cationic uncoupler of oxidative phosphorylation: Interaction with mitochondria detected by derivative spectrophotometry.

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Specific requirement for inorganic phosphate for induction of bilayer membrane conductance by the cationic uncoupler carbocyanine dye

Cited by 17 publications

References 12 publications

Multiple effects of DiS‐C3(5) on mitochondrial structure and function

Multiple effects of DiS‐C3(5) on mitochondrial structure and function

Three-Way Effect of Cyanine Dye on the Structure and Function of Mitochondria

The cyanine dye triS-C4(5) as a cationic uncoupler of oxidative phosphorylation: Interaction with mitochondria detected by derivative spectrophotometry.

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Multiple effects of DiS‐C₃(5) on mitochondrial structure and function

Multiple effects of DiS‐C₃(5) on mitochondrial structure and function