beta cells sense glucose through its metabolism and the resulting increase in ATP, which subsequently stimulates insulin secretion. Uncoupling protein-2 (UCP2) mediates mitochondrial proton leak, decreasing ATP production. In the present study, we assessed UCP2's role in regulating insulin secretion. UCP2-deficient mice had higher islet ATP levels and increased glucose-stimulated insulin secretion, establishing that UCP2 negatively regulates insulin secretion. Of pathophysiologic significance, UCP2 was markedly upregulated in islets of ob/ob mice, a model of obesity-induced diabetes. Importantly, ob/ob mice lacking UCP2 had restored first-phase insulin secretion, increased serum insulin levels, and greatly decreased levels of glycemia. These results establish UCP2 as a key component of beta cell glucose sensing, and as a critical link between obesity, beta cell dysfunction, and type 2 diabetes.

Uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily. Based upon its high homology with UCP1 and its restricted tissue distribution to skeletal muscle and brown adipose tissue, UCP3 has been suggested to play important roles in regulating energy expenditure, body weight, and thermoregulation. Other postulated roles for UCP3 include regulation of fatty acid metabolism, adaptive responses to acute exercise and starvation, and prevention of reactive oxygen species (ROS) formation. To address these questions, we have generated mice lacking UCP3 (UCP3 knockout (KO) mice). Here, we provide evidence that skeletal muscle mitochondria lacking UCP3 are more coupled (i.e. increased state 3/state 4 ratio), indicating that UCP3 has uncoupling activity. In addition, production of ROS is increased in mitochondria lacking UCP3. This study demonstrates that UCP3 has uncoupling activity and that its absence may lead to increased production of ROS. Despite these effects on mitochondrial function, UCP3 does not seem to be required for body weight regulation, exercise tolerance, fatty acid oxidation, or cold-induced thermogenesis. The absence of such phenotypes in UCP3 KO mice could not be attributed to up-regulation of other UCP mRNAs. However, alternative compensatory mechanisms cannot be excluded. The consequence of increased mitochondrial coupling in UCP3 KO mice on metabolism and the possible role of yet unidentified compensatory mechanisms, remains to be determined. Uncoupling protein 3 (UCP3)1 (1-3) is a member of the mitochondrial anion carrier superfamily with high homology (57%) to UCP1, a well characterized uncoupling protein (4, 5). UCP3 together with UCP1, UCP2 (6, 7), and possibly BMCP1 (brain mitochondrial carrier protein) (8) and UCP4 (9), form a family of uncoupling proteins located in the inner mitochondrial membrane. The evidence supporting the uncoupling activity of these proteins comes from studies where UCPs have been heterologously expressed in yeast or reconstituted into proteoliposomes. The expression of UCP2 and -3 decreases the mitochondrial membrane potential, as assessed by uptake of fluorescent membrane potential-sensitive dyes in whole yeast. They also increase state 4 respiration in isolated mitochondria, which serves as an indicator of inner membrane proton leak (3, 6, 10). More recently, reconstitution of UCPs into liposomes has shown that UCP2 and UCP3, like UCP1, mediate proton transport across bilipid layers (11). It is well established that UCP1 is exclusively expressed in brown fat, where it plays a key role in facultative thermogenesis in rodents. Although there is controversy about the molecular mechanisms involved (12-16), it is clear that activated UCP1 catalyzes a proton leak across the mitochondrial inner membrane leading to thermogenesis. The activity of UCP1 is highly regulated, facilitated by fatty acids and inhibited by purine ribose di-and trinucleotides (ATP, ADP, GTP, GDP) (17). UCP1 is also highly regulated at the transcriptional level (18) by cat...

Cells exposed to low oxygen concentrations respond by initiating defense mechanisms, including the stabilization of hypoxia-inducible factor (HIF) 1alpha, a transcription factor that upregulates genes such as those involved in glycolysis and angiogenesis. Nitric oxide and other inhibitors of mitochondrial respiration prevent the stabilization of HIF1alpha during hypoxia. In studies of cultured cells, we show that this effect is a result of an increase in prolyl hydroxylase-dependent degradation of HIF1alpha. With the use of Renilla luciferase to detect intracellular oxygen concentrations, we also demonstrate that, upon inhibition of mitochondrial respiration in hypoxia, oxygen is redistributed toward nonrespiratory oxygen-dependent targets such as prolyl hydroxylases so that they do not register hypoxia. Thus, the signaling consequences of hypoxia may be profoundly modified by nitric oxide.