Measuring Mitochondrial Membrane Potential with a Tetraphenylphosphonium‐Selective Electrode

Moreno, António J.; Santos, Dario L.; Magalhães-Novais, Sílvia; Oliveira, Paulo J.

doi:10.1002/0471140856.tx2505s65

Cited by 13 publications

(9 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The basic physiological studies conducted several decades ago are highly relevant but not always mentioned: Historically, fluorescent dyes [ 18 ] and lipophilic cationic tracers have been developed for quantitative assay of ΔΨ m in isolated mitochondria [ 4 ], cells [ 19 ], and isolated heart preparations [ 6 ]. Electrodes sensitive to tetraphenylphosphonium (TPP) have also been developed and used to evaluate ΔΨ m in isolated mitochondrial fractions [ 9 , 20 ]. [ 14 C or 3 H]-labeled lipophilic cations were used to study the electrical properties of membranes and mitochondria long before modern imaging methods were imagined [ 3 , 21 – 23 ].…”

Section: Introductionmentioning

confidence: 99%

Quantitative in vivo mapping of myocardial mitochondrial membrane potential

et al. 2018

View full text Add to dashboard Cite

BackgroundMitochondrial membrane potential (ΔΨm) arises from normal function of the electron transport chain. Maintenance of ΔΨm within a narrow range is essential for mitochondrial function. Methods for in vivo measurement of ΔΨm do not exist. We use 18F-labeled tetraphenylphosphonium (18F-TPP+) to measure and map the total membrane potential, ΔΨT, as the sum of ΔΨm and cellular (ΔΨc) electrical potentials.MethodsEight pigs, five controls and three with a scar-like injury, were studied. Pigs were studied with a dynamic PET scanning protocol to measure 18F-TPP+ volume of distribution, VT. Fractional extracellular space (fECS) was measured in 3 pigs. We derived equations expressing ΔΨT as a function of VT and the volume-fractions of mitochondria and fECS. Seventeen segment polar maps and parametric images of ΔΨT were calculated in millivolts (mV).ResultsIn controls, mean segmental ΔΨT = -129.4±1.4 mV (SEM). In pigs with segmental tissue injury, ΔΨT was clearly separated from control segments but variable, in the range -100 to 0 mV. The quality of ΔΨT maps was excellent, with low noise and good resolution. Measurements of ΔΨT in the left ventricle of pigs agree with previous in in-vitro measurements.ConclusionsWe have analyzed the factors affecting the uptake of voltage sensing tracers and developed a minimally invasive method for mapping ΔΨT in left ventricular myocardium of pigs. ΔΨT is computed in absolute units, allowing for visual and statistical comparison of individual values with normative data. These studies demonstrate the first in vivo application of quantitative mapping of total tissue membrane potential, ΔΨT.

show abstract

Section: Introductionmentioning

confidence: 99%

Quantitative in vivo mapping of myocardial mitochondrial membrane potential

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Here, ΔΨm represents the central parameter of bioenergetics that has a crucial influence on ATP synthesis in particular, as well as the generation of reactive oxygen species and the respiratory rate [82,83], but is not exclusively responsible for the respiratory rate [84]. A partial improvement of the respiratory chain after RR administration accompanied without an enhanced ΔΨm, therefore, seems plausible.…”

Section: Discussionmentioning

confidence: 99%

“…Following the results of the ΔΨm experiments, RR had no significant influence on intracellular ATP levels, while concentrations were significantly increased by PCA ( Figure 6). Mitochondrial membrane potential is the main driving force for mitochondrial energy production, thus it appears only feasible that improved ΔΨm leads to increased ATP levels [82,83]. Energy-enhancing capacities of already named polyphenols and numerous others have been well documented previously [90,91,92].…”

Section: Discussionmentioning

confidence: 99%

Polyphenols and Metabolites Enhance Survival in Rodents and Nematodes—Impact of Mitochondria

et al. 2019

View full text Add to dashboard Cite

(1) Background: Polyphenols (PP) play an important role in the prevention of non-communicable diseases and may contribute to healthy aging. To investigate the molecular and cellular aspects of PP metabolites on longevity with a focus on mitochondrial function, we applied a pre-fermented mixture of polyphenols (Rechtsregulat®, RR) to rodents and nematodes. (2) Methods: The lifespans of Navar Medical Research Institute (NMRI) mice and C. elegans were recorded. The heat-stress resistance (37 °C) of C. elegans N2 was measured using nucleic staining. Respiration and membrane potential (ΔΨm) were measured in isolated mitochondria. The energetic metabolites adenosine triphosphate (ATP), lactate, and pyruvate were determined in lysates. Expression levels of longevity related genes were determined using quantitative real time polymerase chain reaction (qRT-PCR). Phenolic compounds were identified using ultra high performance liquid chromatography-diode array detection-Iontrap-multiple stage mass spectrometry (UHPLC-DAD-Iontrap-MSn). (3) Results: Several phenolic metabolites including protocatechuic acid (PCA) were identified in RR. Feeding of mice with RR resulted in a significantly increased lifespan. Heat-stress resistance (RR *** p = 0.0006; PCA **** p < 0.0001), median lifespan (NMRI: RR ** p = 0.0035; C. elegans RR * p = 0.0279; PCA **** p < 0.0001), and activity of mitochondrial respiratory chain complexes (RR *−** p = 0.0237 − 0.0052; PCA * p = 0.019 − 0.0208) of C. elegans were significantly increased after incubation with RR (10%) or PCA (780 µM). PCA significantly improved nematodes ΔΨm (* p = 0.02058) and ATP levels (* p = 0.029). RR significantly up-regulated lactate levels, indicating enhanced glycolysis. The expression levels of longevity related genes daf-16, sir-2.1, and skn-1 were significantly upregulated after PCA, and partially after RR administration. (4) Conclusion: Phenolic metabolites such as PCA have the potential to enhance health and lifespan and mitochondrial function, and thus may contribute to healthy aging.

show abstract

“…Tetraphenylphosphonium (TPP + ), a lipophilic cation is widely used as a probe for various cells as well as an artificial membrane potential measurements 20,23–26 . The ability of the aromatic groups to delocalize and shield the electric charge and to increase the lipid solubility facilitates their translocation across the hydrophobic barrier 27,28 .…”

Section: Discussionmentioning

confidence: 99%

“…A custom-build mini-potentiostat with a special high sensitivity electronic circuit, protected against static electrical interference, was used for the measurements 19 . The electrode with the selective membrane was fabricated according to the protocol found in the literature 20 . During all measurements of the changes of the lipophilic tetraphenylphosphonium (TPP + ) ion concentrations, the TPP + chloride (Acros Organics, Belgium) was used.…”

Section: Methodsmentioning

confidence: 99%

The link between yeast cell wall porosity and plasma membrane permeability after PEF treatment

Stirkė

Celiešiūtė-Germanienė

Zimkus³

et al. 2019

Sci Rep

View full text Add to dashboard Cite

An investigation of the yeast cell resealing process was performed by studying the absorption of the tetraphenylphosphonium (TPP+) ion by the yeast Saccharomyces cerevisiae. It was shown that the main barrier for the uptake of such TPP+ ions is the cell wall. An increased rate of TPP+ absorption after treatment of such cells with a pulsed electric field (PEF) was observed only in intact cells, but not in spheroplasts. The investigation of the uptake of TPP+ in PEF treated cells exposed to TPP+ for different time intervals also showed the dependence of the absorption rate on the PEF strength. The modelling of the TPP+ uptake recovery has also shown that the characteristic decay time of the non-equilibrium (PEF induced) pores was approximately a few tens of seconds and this did not depend on the PEF strength. A further investigation of such cell membrane recovery process using a florescent SYTOX Green nucleic acid stain dye also showed that such membrane resealing takes place over a time that is like that occurring in the cell wall. It was thus concluded that the similar characteristic lifetimes of the non-equilibrium pores in the cell wall and membrane after exposure to PEF indicate a strong coupling between these parts of the cell.

show abstract

Measuring Mitochondrial Membrane Potential with a Tetraphenylphosphonium‐Selective Electrode

Cited by 13 publications

References 31 publications

Quantitative in vivo mapping of myocardial mitochondrial membrane potential

Quantitative in vivo mapping of myocardial mitochondrial membrane potential

Polyphenols and Metabolites Enhance Survival in Rodents and Nematodes—Impact of Mitochondria

The link between yeast cell wall porosity and plasma membrane permeability after PEF treatment

Contact Info

Product

Resources

About