PROBING TEMPORAL CHANGES IN MITOCHONDRIAL MEMBRANE POTENTIAL WITH IMPEDANCE SPECTROSCOPY
Chakraborty Mitra, Rooplekha 1971-
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The electrical properties of mitochondria provide fundamental insights into metabolic processes in health and disease. This research studies electrical impedance spectroscopy as a non-invasive, sensitive, and relatively low cost technique to monitor biological processes, such as those involving changes in mitochondrial membrane potential. Our experimental strategy first involves treating suspensions of live mitochondria with the substrate succinate to stimulate activity of succinate dehydrogenase, or more simply Complex II. This triggers electron flux through Complex II and the remaining complexes of the electron transport chain, enabling them to pump protons across the inner membrane and build up a membrane potential. Subsequent variability is introduced by adding various concentrations of the uncoupler trifluorocarbonylcyanide phenylhydrazone (FCCP) and the neurotransmitter dopamine (DA) to mitochondrial suspensions, and measuring changes in impedance. Our results show that adding succinate decreases impedance, consistent with an increase in dielectric response and membrane potential. Overall, our investigation establishes real-time impedance spectroscopy as a non-destructive, potentially powerful method for membrane potential studies of mitochondria.