Monitoring of the mitochondrial and plasma membrane potentials in human fibroblasts by tetraphenylphosphonium ion distribution |
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Authors: | Michela Rugolo Giorgio Lenaz |
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Affiliation: | (1) Laboratorio di Biochimica, Dipartimento di Biologia Evoluzionistica e Sperimentale, Universitá di Bologna, Italy |
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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.Abbreviations m membrane potential across thein situ mitochondria - p membrane potential across the plasma membrane - TPP+ tetraphenylphosphonium - HEPES N-2-hydroxyethylpiperazineN-2-ethanesulfonic acid - FCCP carbonyl cyanidep-trifluoromethoxyphenylhydrazone |
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Keywords: | Human fibroblasts plasma membrane potential mitochondrial membrane potential tetraphenylphosphonium ion |
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