Rat liver mitochondria were found to swell under nonenergized conditions when suspended in media containing 30-40 mM TINO3. Respiration on succinate caused a rapid contraction of mitochondria swollen under nonenergized conditions. In the presence of thallous acetate, there was a rapid initial swelling under nonenergized conditions until a plateau was reached; respiration on succinate then caused a further swelling. Trace amounts of 204Tl (less than 100 microM) equilibrated fairly rapidly across the mitochondrial membrane. The influx of Tl+ was able to promote the decay not only of a valinomycin-induced K+-diffusion potential but also of respiration-generated fields in the inner membrane in accordance with the electrophoretic nature of Tl+ movement. Efflux of Tl+ showed a half-time of about 10 sec at 20 degrees C and was not affected appreciably by the energy state. Efflux was retarded by Mg2+ and by lowering the temperature. The data indicate that Tl+ when present at high concentrations, 30mM or more, distributes across the mitochondrial inner membrane both in response to electrical fields and to delta pH. In energized mitochondria the uptake of Tl+ would occur electrophoretically, while Tl+/H+ exchange would constitute a leak. In the presence of NO3-, the movements of Tl+ are determined by that of NO3-, indicating short-range coupling of electrical forces. At low concentrations of Tl+, 5 mM or less, there was no indication of a Tl+/H+ exchange, which appears to be induced by high concentrations of Tl+.

Nonrespiring rat-liver mitochondria swell in media containing high concentrations of thallous nitrate, indicating passive penetration of Tl'. This swelling could be further stimulated by 10 nM or more nonactin while even 1 pM valinomycin was without effect. Nonactin was also much more potent than valinomycin in stimulating swelling of respiring mitochondria in the presence of thallous acetate. It is evident that nonactin acts as a potent ionophore of T1' able to promote both the passive and energized uptake of T1' in mitochondria.The distribution of TI+, present in trace concentrations below 1 mM, was measured during energisation by respiration both in the presence and absence of ionophores. Respiration induced net uptake of T1+ only in the presence of ionophores, though T1+ as a permeant cation was expected to sense respiration-induced changes in the membrane potential. The data may be interpreted as indicating that no transmembrane potential is formed upon energisation, but localized fields, which are able to interact with the lipophilic ionophore complexes of TI +, but not with the hydrophilic cation Tl'. This interpretation is valid only if thermodynamic equilibrium has been reached.

The distribution of T1+ between rat liver mitochondria and the medium was studied; millimolar or smaller concentrations of T1+ were labeled with 204T1. The T1+ distribution responded to transient diffusion potentials in a way that indicated electrophoretic movements of T1+. The diffusion potentials were induced by efflux of K+ in response to addition of valinomycin to nonrespiring mitochondria suspended in a medium with low concentrations of K+ or by efflux of H+ induced by making the medium more alkaline in the presence of a protonophorous (proton-conducting) uncoupling agent. Changes in membrane potential induced by valinomycin were followed with the aid of safranine. T1+ brought about collapse of the diffusion potential. It is concluded that T1+ is able to penetrate the mitochondrial membrane electrophoretically.

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