Abstract
The lipophilic cation tetraphenylphosphonium (TPP+) is accumulated by human skin fibroblasts across both the plasma and mitochondrial membranes. We show here that TPP+ uptake is indeed greatly decreased under conditions leading to de-energization of mitochondria. The TPP+ accumulation in the presence of the proton ionophore FCCP has been used for determination of the plasma membrane potential across the plasma membrane, after correction for potential-independent binding of TPP+ to cellular components. Following this procedure, a value of 75 mV has been obtained. Through the amount of TPP+ released by FCCP treatment, an estimate of thein situ mitochondrial membrane potential has been made. Furthermore, we report that the mitochondrial component of TPP+ accumulation decreases with aging of fibroblast cultures.
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Abbreviations
- ΔΨm :
-
membrane potential across thein situ mitochondria
- ΔΨp :
-
membrane potential across the plasma membrane
- TPP+ :
-
tetraphenylphosphonium
- HEPES:
-
N-2-hydroxyethylpiperazineN′-2-ethanesulfonic acid
- FCCP:
-
carbonyl cyanidep-trifluoromethoxyphenylhydrazone
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Rugolo, M., Lenaz, G. Monitoring of the mitochondrial and plasma membrane potentials in human fibroblasts by tetraphenylphosphonium ion distribution. J Bioenerg Biomembr 19, 705–718 (1987). https://doi.org/10.1007/BF00762304
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DOI: https://doi.org/10.1007/BF00762304