Patch clamping the outer mitochondrial membrane

Abstract

Intact giant mitochondria isolated from the liver of mice fed a diet containing cuprizone were studied using patch microelectrodes. The current-voltage curves were nonlinear, suggesting the presence of voltage-sensitive channels. In the negative range of voltage, the channels appear to close with increasing magnitude of the voltage. The dependence of the conductance on voltage is similar to that of the outer membrane channels (VDAC) studied in planar bilayers. Occasionally, over a narrow range of positive potentials, the conductance also decreases as in the bilayer studies. However, more frequently the conductance increases sharply in a completely reversible manner at potentials greater than 10 to 20 mV. The increase in conductance with voltage may be interpreted as a major rearrangement of membrane components. Qualitatively comparable results were obtained using fused outer membranes isolated fromNeurospora mitochondria. The behavior of VDAC is affected by treatment with succinic anhydride or the polyanion, polymethacrylate, maleate, styrene (1∶2∶3). We have found similar effects in the negative range of potentials in patches from giant mitochondria treated in the same fashion.