In contrast to UCP1, the primary function of UCP3 is not the dissipation of energy. Rather, several lines of evidence suggest that UCP3 is related to cellular long-chain fatty acid homeostasis. If long-chain fatty acids enter the mitochondrial matrix in their non-esterified form, they cannot be metabolized and may exert deleterious effects. To test the feasibility that UCP3 exports fatty acid anions, we systematically interfered at distinct steps in the fatty acid metabolism pathway, thereby creating conditions in which the entry of (non-esterified) fatty acids into the mitochondrial matrix is enhanced. First, reducing the cellular fatty acid binding capacity, known to increase cytosolic concentrations of non-esterified fatty acids, up-regulated UCP3 5.3-fold. Second, inhibition of mitochondrial entry of esterified long-chain fatty acids upregulated UCP3 by 1.9-fold. Third, high-fat diets, to increase mitochondrial supply of nonesterified long-chain fatty acids exceeding oxidative capacity, up-regulated UCP3 twofold. However, feeding a similar amount of medium-chain fatty acids, which can be oxidized inside the mitochondrial matrix and therefore do not need to be exported from the matrix, did not affect UCP3 protein levels. These data are compatible with a physiological function of UCP3 in facilitating outward transport of long-chain fatty acid anions, which cannot be oxidized, from the mitochondrial matrix.Key words: UCP3 • fatty acid transport • mitochondria • high-fat diet n contrast to the brown adipose tissue-specific UCP1, there is ample evidence that the physiological function of UCP3 is not in energy dissipation. For example, fasting, an energy preserving condition, rapidly up-regulates UCP3 (1). We have recently demonstrated in humans that a diet-induced up-regulation of UCP3 did not affect mitochondrial coupling, suggesting that the primary function of UCP3 is not mitochondrial uncoupling (2). Rather, several lines of evidence suggest that UCP3 is related to cellular fatty acid metabolism. Thus, I aberrations in whole-body fat oxidation were observed in humans with an exon-6 splice donor mutation in UCP3 (3), as well as in UCP3 knockout mice (4). Furthermore, in skeletal muscle UCP3 is rapidly up-regulated during fasting (1), acute exercise (5, 6), and high dietary fat-intake (7,8), all situations in which fat metabolism is affected. In addition, UCP3 protein content declines in situations in which fat oxidative capacity is improved, such as after endurance training (9, 10) and after weight reduction (11,12), and UCP3 has lowest expression in Type 1 muscle fibers, which are characterized by a high fat oxidative capacity (13,14).This pattern of regulation of UCP3 can consistently be explained by considering the balance between fatty acid delivery to mitochondria and their capacity to oxidize fatty acids. In situations characterized by a positive balance (i.e., fatty acid delivery exceeds oxidative capacity), such as fasting, high-fat intake, acute exercise, and in Type 2b fibers, UCP3 levels are...

Human and rodent uncoupling protein (UCP)3 mRNA is upregulated after acute exercise. Moreover, exercise increases plasma levels of free fatty acid (FFA), which are also known to upregulate UCP3. We investigated whether the upregulation of UCP3 after exercise is an effect of exercise per se or an effect of FFA levels or substrate oxidation. Seven healthy untrained men [age: 22.7 +/- 0.6 yr; body mass index: 23.8 +/- 1.0 kg/m(2); maximal O2 uptake (VO2 max): 3,852 +/- 211 ml/min] exercised at 50% VO2 max for 2 h and then rested for 4 h. Muscle biopsies and blood samples were taken before and immediately after 2 h of exercise and 1 and 4 h in the postexercise period. To modulate plasma FFA levels and fat/glucose oxidation, the experiment was performed two times, one time with glucose ingestion and one time while fasting. UCP3 mRNA and UCP3 protein were determined by RT-competitive PCR and Western blot. In the fasted state, plasma FFA levels significantly increased (P < 0.0001) during exercise (293 +/- 25 vs. 1,050 +/- 127 micromol/l), whereas they were unchanged after glucose ingestion (335 +/- 54 vs. 392 +/- 74 micromol/l). Also, fat oxidation was higher after fasting (P < 0.05), whereas glucose oxidation was higher after glucose ingestion (P < 0.05). In the fasted state, UCP3L mRNA expression was increased significantly (P < 0.05) 4 h after exercise (4.6 +/- 1.2 vs. 9.6 +/- 3.3 amol/microg RNA). This increase in UCP3L mRNA expression was prevented by glucose ingestion. Acute exercise had no effect on UCP3 protein levels. In conclusion, we found that acute exercise had no direct effect on UCP3 mRNA expression. Abolishing the commonly observed increase in plasma FFA levels and/or fatty acid oxidation during and after exercise prevents the upregulation of UCP3 after acute exercise. Therefore, the previously observed increase in UCP3 expression appears to be an effect of prolonged elevation of plasma FFA levels and/or increased fatty acid oxidation.

Uncoupling protein 3 (UCP3) is a muscle mitochondrial protein believed to uncouple the respiratory chain, producing heat and reducing aerobic ATP production. Our aim was to quantify and compare the UCP3 protein levels in type I, IIa and IIx skeletal muscle fibers of endurance-trained (Tr) and healthy untrained (UTr) individuals. UCP3 protein content was quantified using Western blot and immunofluorescence. Skeletal muscle fiber type was determined by both an enzymatic ATPase stain and immunofluorescence. UCP3 protein expression measured in skeletal muscle biopsies was 46% lower ( P=0.01) in the Tr compared to the UTr group. UCP3 protein expression in the different muscle fibers was expressed as follows; IIx>IIa>I in the fibers for both groups ( P<0.0167) but was lower in all fiber types of the Tr when compared to the UTr subjects ( P<0.001). Our results show that training status did not change the skeletal muscle fiber hierarchical UCP3 protein expression in the different fiber types. However, it affected UCP3 content more in type I and type IIa than in the type IIx muscle fibers. We suggest that this decrease may be in relation to the relative improvement in the antioxidant defense systems of the skeletal muscle fibers and that it might, as a consequence, participate in the training induced improvement in mechanical efficiency.

Background: We investigated whether substituting sitting with standing and self-perceived light walking in free-living conditions would improve cardiometabolic risk factors, mood, and cognition in overweight/obese adults.Methods: In a randomized, cross-over study, 24 (m/f: 13/11) sedentary overweight/obese participants (64 ± 7 years, BMI 29 ± 2 kg/m2) followed two activity regimens of each 4 days in free-living conditions: “Sit”: sitting 13.5 h/day, standing 1.4 h/day, self-perceived light-intensity walking 0.7 h/day; for “SitLess” these activities lasted 7.6, 4.0, and 4.3 h/day, respectively. Meals were standardized and physical activity was assessed by accelerometry (activPAL). Insulin sensitivity (expressed as Matsuda-index based on an oral glucose tolerance test), circulating lipids, blood pressure, mood (pleasantness and arousal), and cognition were assessed on the morning after the activity regimens. Quality of life and sleep were assessed on the last day of the activity regimens.Results: We observed that AUC (0–190 min) for insulin decreased by 20% after SitLess vs. Sit [10,125 (656) vs. 12,633 (818); p = 0.006]. Insulin sensitivity improved by 16% after SitLess vs. Sit [Matsuda-index, mean (SEM): 6.45 (0.25) vs. 5.58 (0.25) respectively; p = 0.007]. Fasting triglycerides, non-HDL-cholesterol, and apolipoprotein B decreased by 32, 7, and 4% respectively, whereas HDL-cholesterol increased by 7% after SitLess vs. Sit (all p < 0.01). Diastolic blood pressure was lower after SitLess vs. Sit (p < 0.05). Pleasantness (as one marker of mood status) after the oral glucose tolerance test was higher after SitLess vs. Sit (p < 0.05). There was no significant difference between regimens for cognition, quality of life and sleep.Conclusions: Reducing sitting time in free-living conditions markedly improved insulin sensitivity, circulating lipids, and diastolic blood pressure. Substituting sitting with standing and self-perceived light walking is an effective strategy to improve cardiometabolic risk factors in overweight/obese subjects.

The physiological function of human uncoupling protein-3 is still unknown. Uncoupling protein-3 is increased during fasting and high-fat feeding. In these situations the availability of fatty acids to the mitochondria exceeds the capacity to metabolize fatty acids, suggesting a role for uncoupling protein-3 in handling of non-metabolizable fatty acids. To test the hypothesis that uncoupling protein-3 acts as a mitochondrial exporter of non-metabolizable fatty acids from the mitochondrial matrix, we gave human subjects Etomoxir (which blocks mitochondrial entry of fatty acids) or placebo in a cross-over design during a 36-h stay in a respiration chamber. Etomoxir inhibited 24-h fat oxidation and fat oxidation during exercise by ~14-19%. Surprisingly, uncoupling protein-3 content in human vastus lateralis muscle was markedly upregulated within 36 h of Etomoxir administration. Up-regulation of uncoupling protein-3 was accompanied by lowered fasting blood glucose and increased translocation of glucose transporter-4. These data support the hypothesis that the physiological function of uncoupling protein-3 is to facilitate the outward transport of non-metabolizable fatty acids from the mitochondrial matrix and thus prevents mitochondria from the potential deleterious effects of high fatty acid levels. In addition our data show that up-regulation of uncoupling protein-3 can be beneficial in the treatment of type 2 diabetes.Key words: fat oxidation • human • GLUT4 • translocation T he human uncoupling protein-3 (UCP3) is a recently identified member of the uncoupling protein family, which is expressed primarily in skeletal muscle. The physiological function of UCP3 is still unresolved (1, 2). Due to its homology with the brown adipose tissuespecific uncoupling protein-1 (UCP1), UCP3 was suggested to be involved in human energy turnover and obesity (3,4). However, mice lacking UCP3 have a normal metabolic rate and body weight (5, 6), and fasting, an energy preserving condition, rapidly up-regulates the expression of UCP3 (7), contradicting a role for UCP3 in the regulation of energy turnover. Rather, UCP3 protein content is consistently up-regulated in situations in which fatty acid delivery to skeletal muscle exceeds the muscle's fat oxidative capacity, such as fasting, acute exercise, and high-fat feeding (7-9). On the other hand, a reduction of UCP3 protein content is observed in situations in which fat oxidative capacity is improved, such as after endurance training (10) and after weight reduction (11). Finally, we showed that UCP3 protein content is highest in type 2b muscle fibers, which are characterized by a low capacity to oxidize fatty acids and therefore are unable to metabolize all cytosolic fatty acids (12). Together with the finding that UCP3 can transport fatty acid anions (13), these results suggest that UCP3 might be involved in the mitochondrial transport of non-metabolizable fatty acids (14).The majority of the fatty acids in the cytoplasm are converted to their esterified form (fatty acylCoA) ...