A PVC-based electrode sensitive to DDA+ as a device for monitoring the membrane potential in biological systems

A polyvinyl chloride (PVC)-based membrane electrode sensitive to dibenzyldimethyl ammonium cation (DDA⁺) was constructed, and operational parameters such as the selectivity coefficients, the detection limit, and the response time were obtained. In comparison with the selectivity coefficients obtained with the previous liquid-membrane electrode, significant improvement was not obtained, but the response time became pronouncedly shorter. Furthermore, the electrode lifetime was remarkably prolonged. With the electrode developed, the change in the membrane potential of liposomes containing dibutyl ferrocene which separated oxidizing and reducing agent solutions was measured. The DDA⁺ uptake, U, and the membrane potential estimated from U changed in accordance with the redox potential in the medium when the concentration of internal ferricyanide was kept constant. The membrane potential collapsed when the uncoupler of oxidative phosphorylation was added. The ANS fluorescence measurement indicated that negative charges appeared on energization with oxidizing-reducing agent. The change in membrane potential of mitochondria during energization was also measured. It was found that the liposome described above is a good model for the generation of membrane potentials in mitochondria.