The use of the uncoupling protein inhibitor GDP was used to investigate UCP 1 dependent ROS production in BAT and thymus mitochondria. The effect of UCP 1 inhibition by GDP was then corroborated using UCP 1 ablated mice and their wild-type controls. It was shown that UCP 1 was capable of attenuating ROS production in isolated BAT mitochondria. ROS due to reverse electron transport was shown to be particularly sensitive to UCP 1 attenuation. We have shown for the first time that BAT mitochondria from UCP 1 knock-out mice have significantly less complex I activity and abundance, thereby reducing ROS production due to reverse electron transport, and possibly explaining previous reports of no difference in oxidative damage in BAT mitochondria from UCP 1 knock-out and wildtype mice. We also show for the first time that BAT mitochondria from UCP 1 knock-out mice, surprisingly, have significantly greater glycerol- 3-phosphate dehydrogenase activity.

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