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Fig. 1. Typical output from a TPP+-sensitive electrode during
calibration and the measurement of membrane potential of liver mitochondria
isolated from a summer active ground squirrel. Mitochondria (1 mg protein)
were incubated with rotenone (5 µmoll–1), nigericin (80 ng
ml–1) and oligomycin (1 µg ml–1) after
which TPP+ electrodes were calibrated by five additions of
TPP+, each of which raised [TPP+] by 1
µmoll–1. Addition of succinate initiated
nonphosphorylating respiration, which was subsequently inhibited with five
additions of malonate (0.5 µmoll–1). Oxygen consumption
was monitored simultaneously. FCCP was added at the conclusion of the assay to
uncouple respiration completely, releasing unbound TPP+ from the
mitochondrial matrix and allowing for correction of electrode drift.
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