Mitochondrial Oxidative Phosphorylation Efficiency Research Articles

Meat-type chickens have a higher efficiency of mitochondrial oxidative phosphorylation than laying-type chickens

Meat-type chickens show high feed efficiency and have a very rapid growth rate compared with laying-type chickens. To clarify whether the type-specific difference in feed conversion efficiency is involved in mitochondrial bioenergetics, modular kinetic analysis was applied to oxidative phosphorylation in skeletal muscle mitochondria of both type chickens. Mitochondria from skeletal muscle of meat-type chickens showed greater substrate oxidation and phosphorylating activities, and less proton leak than those of the laying-type, resulting in a higher efficiency of oxidative phosphorylation. Gene expression and protein content of uncoupling protein (avUCP) but not adenine nucleotide translocase (avANT) gene expression were lower in skeletal muscle mitochondria of meat-type chickens than the laying-type. The current results regarding a higher efficiency of oxidative phosphorylation and UCP content may partially support the high feed efficiency of meat-type chickens.

Tissue variation of mitochondrial oxidative phosphorylation efficiency in cold-acclimated ducklings.

We investigated the oxidative phosphorylation efficiency of liver and gastrocnemius muscle mitochondria in thermoneutral and cold-acclimated ducklings. The yield of oxidative phosphorylation was lower in muscle than in liver mitochondria, a difference that was associated with a higher proton conductance in muscle mitochondria. Cold exposure did not affect oxidative phosphorylation efficiency or basal proton leak in mitochondria. We conclude that the basal proton conductance of mitochondria may regulate mitochondrial oxidative phosphorylation efficiency, but is not an important contributor to thermogenic processes in cold-acclimated ducklings.

Mitochondrial Dysfunction Contributes to Impaired Insulin Secretion in INS-1 Cells with Dominant-negative Mutations of HNF-1α and in HNF-1α-deficient Islets

Maturity Onset Diabetes of the Young-type 3 (MODY-3) has been linked to mutations in the transcription factor hepatic nuclear factor (HNF)-1alpha, resulting in deficiency in glucose-stimulated insulin secretion. In INS-1 cells overexpressing doxycycline-inducible HNF-1alpha dominant-negative (DN-) gene mutations, and islets from Hnf-1alpha knock-out mice, insulin secretion was impaired in response to glucose (15 mm) and other nutrient secretagogues. Decreased rates of insulin secretion in response to glutamine plus leucine and to methyl pyruvate, but not potassium depolarization, indicate defects specific to mitochondrial metabolism. To identify the biochemical mechanisms responsible for impaired insulin secretion, we used (31)P NMR measured mitochondrial ATP synthesis (distinct from glycolytic ATP synthesis) together with oxygen consumption measurements to determine the efficiency of mitochondrial oxidative phosphorylation. Mitochondrial uncoupling was significantly higher in DN-HNF-1alpha cells, such that rates of ATP synthesis were decreased by approximately one-half in response to the secretagogues glucose, glutamine plus leucine, or pyruvate. In addition to closure of the ATP-sensitive K(+) channels with mitochondrial ATP synthesis, mitochondrial production of second messengers through increased anaplerotic flux has been shown to be critical for coupling metabolism to insulin secretion. (13)C-Isotopomer analysis and tandem mass spectrometry measurement of Krebs cycle intermediates revealed a negative impact of DN-HNF-1alpha and Hnf-1alpha knock-out on mitochondrial second messenger production with glucose but not amino acids. Taken together, these results indicate that, in addition to reduced glycolytic flux, uncoupling of mitochondrial oxidative phosphorylation contributes to impaired nutrient-stimulated insulin secretion with either mutations or loss of HNF-1alpha.

Computational Analysis of Cardiac Energetics during Ischemia and Reperfusion in Buffer‐Perfused Rabbit Hearts

To determine the mechanisms by which nucleotide concentrations change in the heart during ischemia and reperfusion, data from Kroll et al. [Am. J. Physiol. 272:H2567‐2576, 1997] were analyzed using a computational model of cardiac energy metabolism and metabolite transport. In the experiments of Kroll et al., rabbit hearts were subjected to 10 minutes of perfusion at basal conditions; then 45 minutes of ischemia with 95% flow reduction; and then reperfusion. Phosphate metabolite concentrations were measured by 31P‐Magnetic Resonance Spectroscopy (31P‐MRS) and oxygen flux by arterial‐venous differences in oxygen tension during the ischemia reperfusion protocol. Data were analyzed by integrating open adenosine kinetics into a previously developed computational model [Wu et al., J. Physiol. 586:4193‐4208, 2008]. Our analyses find that during ischemia/anoxia the heart actively down‐regulates ATP hydrolysis rate in the cytoplasm, while inhibiting mitochondrial ATP consumption. Down‐regulation of mitochondria ATP consumption during anoxia may be associated with inhibition of F1Fo‐ATPase reversal or reduction of mitochondrial proton leak. During the reperfusion period, the mitochondrial ATP synthesis capacity is largely restored, while mitochondrial proton leak is significantly magnified resulting in a slightly impaired efficiency of mitochondrial oxidative phosphorylation. The loss of adenosine nucleotides during ischemia does not limit the mitochondrial oxidative capacity during reperfusion.

The Efficiency of Cellular Energy Transduction and Its Implications for Obesity

We assess the existence, mechanism, and functions of less-than-maximal coupling efficiency of mitochondrial oxidative phosphorylation and its potential as a target for future antiobesity interventions. Coupling efficiency is the proportion of oxygen consumption used to make adenosine triphosphate (ATP) and do useful work. High coupling efficiency may lead to fat deposition; low coupling efficiency to a decrease in fat stores. We review obligatory and facultative energy expenditure and the role of a futile cycle of proton pumping and proton leak across the mitochondrial inner membrane in dissipating energy. Basal proton conductance is catalyzed primarily by the adenine nucleotide translocase but can be mimicked by chemical uncouplers. Inducible proton conductance is catalyzed by specific uncoupling proteins. We discuss the opportunities and pitfalls of targeting these processes as a treatment for obesity by decreasing coupling efficiency and increasing energy expenditure, either directly or through central mechanisms of energy homeostasis.

Metformin induces oxidative stress in white adipocytes and raises uncoupling protein 2 levels

Metformin is a drug widely used to treat type 2 diabetes. It enhances insulin sensitivity by improving glucose utilization in tissues like liver or muscle. Metformin inhibits respiration, and the decrease in cellular energy activates the AMP-activated protein kinase that in turn switches on catabolic pathways. Moreover, metformin increases lipolysis and beta-oxidation in white adipose tissue, thereby reducing the triglyceride stores. The uncoupling proteins (UCPs) are transporters that lower the efficiency of mitochondrial oxidative phosphorylation. UCP2 is thought to protect against oxidative stress although, alternatively, it could play an energy dissipation role. The aim of this work was to analyse the involvement of UCP2 on the effects of metformin in white adipocytes. We studied the effect of this drug in differentiating 3T3-L1 adipocytes and found that metformin causes oxidative stress since it increases the levels of reactive oxygen species (ROS) and lowers the aconitase activity. Variations in UCP2 protein levels parallel those of ROS. Metformin also increases lipolysis in these cells although only when the levels of ROS and UCP2 have decreased. Hence, UCP2 does not appear to be needed to facilitate fatty acid oxidation. Furthermore, treatment of C57BL/6 mice with metformin also augmented the levels of UCP2 in epididymal white adipose tissue. We conclude that metformin treatment leads to the overexpression of UCP2 in adipocytes to minimize the oxidative stress that is probably due to the inhibition of respiration caused by the drug.

Genomic modulation of mitochondrial respiratory genes in the hypertrophied heart reflects adaptive changes in mitochondrial and contractile function

We hypothesized the coordinate induction of mitochondrial regulatory genes in the hypertrophied right ventricle to sustain mitochondrial respiratory capacity and contractile function in response to increased load. Wistar rats were exposed to hypobaric hypoxia (11% O(2)) or normoxia for 2 wk. Cardiac contractile and mitochondrial respiratory function were separately assessed for the right and left ventricles. Transcript levels of several mitochondrial regulators were measured. A robust hypertrophic response was observed in the right (but not left) ventricle in response to hypobaric hypoxia. Mitochondrial O(2) consumption was increased in the right ventricle, while proton leak was reduced vs. normoxic controls. Citrate synthase activity and mitochondrial DNA content were significantly increased in the hypertrophied right ventricle, suggesting higher mitochondrial number. Transcript levels of nuclear respiratory factor-1, peroxisome proliferator-activated receptor-gamma-coactivator-1alpha, cytochrome oxidase (COX) subunit II, and uncoupling protein-2 (UCP2) were coordinately induced in the hypertrophied right ventricle following hypoxia. UCP3 transcript levels were significantly reduced in the hypertrophied right ventricle vs. normoxic controls. Exposure to chronic hypobaric hypoxia had no significant effects on left ventricular mitochondrial respiration or contractile function. However, COXIV and UCP2 gene expression were increased in the left ventricle in response to chronic hypobaric hypoxia. In summary, we found coordinate induction of several genes regulating mitochondrial function and higher mitochondrial number in a model of physiological right ventricular hypertrophy, linking the efficiency of mitochondrial oxidative phosphorylation and respiratory function to sustained contractile function in response to the increased load.

Insulin protects against amyloid β-peptide toxicity in brain mitochondria of diabetic rats

This study compared the status of brain mitochondria isolated from 12-week streptozotocin (STZ)-diabetic rats versus STZ-diabetic animals treated with insulin during a period of 4 weeks. Brain mitochondria isolated from 12-week citrate (vehicle)-treated rats were used as control. For that purpose, several mitochondrial parameters were evaluated: respiratory indexes (respiratory control ratio (RCR) and ADP/O ratio), transmembrane potential (DeltaPsim), repolarization lag phase, repolarization level, ATP, glutathione and coenzyme Q (CoQ) contents, production of H2O2, ATPase activity, and the capacity of mitochondria to accumulate Ca2+. Furthermore, the effect of Abeta1-40 was also analyzed. We observed that STZ-induced diabetes promoted a significant decrease in mitochondrial CoQ9, ATPase activity, and a lower capacity of mitochondria to accumulate Ca2+ when compared with control and insulin-treated diabetic rats. The presence of 4 microM Abeta1-40 induced a significant decrease in RCR in the three groups of rats. However, this peptide induced a significant increase in the repolarization lag phase and a significant decrease in the repolarization level in control and diabetic animals without insulin treatment. Furthermore, this peptide exacerbated significantly the production of H2O2 in STZ-diabetic rats, this effect being avoided by insulin treatment. Our data show that although diabetes induces some alterations in brain mitochondrial activity, those alterations do not interfere significantly with mitochondria functional efficiency. Similarly, insulin does not affect basal mitochondria function. However, in the presence of amyloid beta-peptide, insulin seems to prevent the decline in mitochondrial oxidative phosphorylation efficiency and avoids an increase in oxidative stress, improving or preserving the function of neurons under adverse conditions, such as Alzheimer's disease.

Melatonin as an antioxidant: biochemical mechanisms and pathophysiological implications in humans.

This brief resume enumerates the multiple actions of melatonin as an antioxidant. This indoleamine is produced in the vertebrate pineal gland, the retina and possibly some other organs. Additionally, however, it is found in invertebrates, bacteria, unicellular organisms as well as in plants, all of which do not have a pineal gland. Melatonin's functions as an antioxidant include: a), direct free radical scavenging, b), stimulation of antioxidative enzymes, c), increasing the efficiency of mitochondrial oxidative phosphorylation and reducing electron leakage (thereby lowering free radical generation), and 3), augmenting the efficiency of other antioxidants. There may be other functions of melatonin, yet undiscovered, which enhance its ability to protect against molecular damage by oxygen and nitrogen-based toxic reactants. Numerous in vitro and in vivo studies have documented the ability of both physiological and pharmacological concentrations to melatonin to protect against free radical destruction. Furthermore, clinical tests utilizing melatonin have proven highly successful; because of the positive outcomes of these studies, melatonin's use in disease states and processes where free radical damage is involved should be increased.

Melatonin as an antioxidant: biochemical mechanisms and pathophysiological implications in humans.

This brief resume enumerates the multiple actions of melatonin as an antioxidant. This indoleamine is produced in the vertebrate pineal gland, the retina and possibly some other organs. Additionally, however, it is found in invertebrates, bacteria, unicellular organisms as well as in plants, all of which do not have a pineal gland. Melatonin's functions as an antioxidant include: a), direct free radical scavenging, b), stimulation of antioxidative enzymes, c), increasing the efficiency of mitochondrial oxidative phosphorylation and reducing electron leakage (thereby lowering free radical generation), and 3), augmenting the efficiency of other antioxidants. There may be other functions of melatonin, yet undiscovered, which enhance its ability to protect against molecular damage by oxygen and nitrogen-based toxic reactants. Numerous in vitro and in vivo studies have documented the ability of both physiological and pharmacological concentrations to melatonin to protect against free radical destruction. Furthermore, clinical tests utilizing melatonin have proven highly successful; because of the positive outcomes of these studies, melatonin's use in disease states and processes where free radical damage is involved should be increased.

Correlation between decreased expression of mitochondrial F0F1-ATP synthase and low regenerating capability of the liver after partial hepatectomy in hypothyroid rats.

In hypothyroid rats, partial hepatectomy does not induce liver regeneration until 120 h after surgical operation. when, instead, in normal rats a complete recovery of the liver mass, in this interval, is observed. In normal rats, a good efficiency of mitochondrial oxidative phosphorylation is needed as an energy source for liver regeneration (Guerrieri, F. et al., 1995); in hypothyroid rats the efficiency of mitochondrial oxidative phosphorylation is low in the 0-120 h interval after partial hepatectomy. This low efficiency of oxidative phosphorylation appears to be related to a low mitochondrial content of F0F1-ATP synthase, in liver of hypothyroid rats, which does not recover after partial hepatectomy. In the liver of hypothyroid rats, low levels of the nuclear-encoded mitochondrial catalytic betaF1 subunit and of its transcript are observed and they do not increase, as occurs in normal rats, after partial hepatectomy.

EGb 761 protects liver mitochondria against injury induced by in vitro anoxia/reoxygenation

The present study investigated the protective effects of Ginkgo biloba extract (EGb 761) on rat liver mitochondrial damage induced by in vitro anoxia/reoxygenation. Anoxia/reoxygenation was known to impair respiratory activities and mitochondrial oxidative phosphorylation efficiency. ADP/O (2.57 ± 0.11) decreased after anoxia/reoxygenation (1.75 ± 0.09, p < .01), as well as state 3 and uncoupled respiration (−20%, p < .01), but state 4 respiration increased ( p < .01). EGb 761 (50–200 μg/ml) had no effect on mitochondrial functions before anoxia, but had a specific dose-dependent protective effect after anoxia/reoxygenation. When mitochondria were incubated with 200 μg/ml EGb 761, they showed an increase in ADP/O (2.09 ± 0.14, p < .05) and a decrease in state 4 respiration (−22%) after anoxia/reoxygenation. In EPR spin-trapping measurement, EGb 761 decreased the EPR signal of superoxide anion produced during reoxygenation. In conclusion, EGb 761 specially protects mitochondrial ATP synthesis against anoxia/reoxygenation injury by scavenging the superoxide anion generated by mitochondria.

Efficiency of oxidative work performance of skeletal muscle in patients with cystic fibrosis.

Exercise intolerance in patients with cystic fibrosis is commonly attributed to reduced pulmonary and nutritional status. The possible role of diminished efficiency of mitochondrial oxidative phosphorylation in relation to skeletal muscle performance was investigated in patients with cystic fibrosis. In vivo synthesis of ATP in skeletal muscle during submaximal exercise was studied in eight patients with cystic fibrosis aged 12-17 years, and in 19 healthy control subjects aged 8-36 years. The intracellular pH and concentrations of phosphate compounds were calculated at four steady states from phosphorus-31 labelled nuclear magnetic resonance spectroscopy measurements in the forearm muscle during bulb squeezing in an exercise protocol. Normalised power output, expressed as percentage maximal voluntary contraction (Y, in %MVC), was related to the energy force of ATP hydrolysis (X = ln [ATP]/[ADP][Pi]). This relationship provides an in vivo measure of efficiency of oxidative work performance of skeletal muscle. During all workloads (but not at rest) intracellular pH was higher in the patients with cystic fibrosis than in the controls. The linear least square fit for Y = a-bX showed high correlations in both groups; the slope b was 19% lower in the patients than in the controls (11.8% v 14.5% MVC/ln M; 95% confidence interval for difference 0.3 to 5.0). In patients with cystic fibrosis oxidative work performance of skeletal muscle is reduced. This may be related to secondary pathophysiological changes in skeletal muscle in cystic fibrosis.

Energetic behaviour of mitochondria isolated from rat livers perfused with a perfluorodecalin + N, N-perfluorodiethylcyclohexylamine (emulsion)

Rat livers have been perfused with a saline control medium or with a perfluorocarbon emulsion containing perfluorodecalin and N,N-perfluorodiethylcyclohexylamine, and the respiratory rates and transmembrane electrical potentials of mitochondria isolated following perfusion have been evaluated. The results indicate that the perfluorocarbon emulsion used, by providing a good oxygen supply to the perfused liver, allowed to preserve the efficiency of mitochondrial oxidative phosphorylation.

The energy equivalents of ATP and the energy values of food proteins and fats.

Heats of combustion and energy equivalents of cytoplasmic ATP have been estimated for glucose, 101 food proteins and 116 food fats based on amino acid and fatty acid composition data from food composition tables and the heats of combustion and energy equivalents of cytoplasmic ATP of each individual amino acid, fatty acid, glycerol and glucose. The isodynamic equivalents of carbohydrate, fat and protein at the biochemical level have been investigated. Heats of combustion of food proteins and fats derived from compositional data were within 1% of published values obtained by calorimetry. Cytoplasmic ATP equivalents for glucose, fat and protein range from 9.0 to 14.7, 8.6 to 14.6 and 6.4 to 13.2 mol cytoplasmic ATP/MJ of metabolizable energy respectively, depending on the choice of mitochondrial proton stoichiometries for these estimations. The range is extended further when considering the level and type of mitochondrial 'uncoupling'. Isobioenergetic relationships between the efficiencies of glucose (G) and fat (F) (F = 1.05 G - 0.9) and glucose and protein (P) (P = G(1.02 - 0.19 f) - (1.8 + 0.5 f] energy conversions (where f is the fraction of protein oxidized via gluconeogenesis) were obtained and were essentially independent of the choice of mitochondrial proton stoichiometry and the level and type of uncoupling of oxidative phosphorylation. Potential errors in previous estimates of ATP yield from protein are shown to be as much as -17.6 to greater than 118%; accounting for the efficiency of mitochondrial oxidative phosphorylation narrows this to between -7.9 and 17.4% and accounting for the fraction of protein oxidized via gluconeogenesis limits this further to between -7.9 and 11.1%. Remaining uncertainty is attributed mostly to lack of knowledge about the energy cost of substrate absorption from the gut and transport across cell membranes. Coefficients of variation (cv) in the cytoplasmic ATP yield/g protein and /g protein nitrogen for the 101 food proteins were large (0.033 and 0.058 respectively). This is attributed mostly to variation in the metabolizable heats of combustion (cv 0.033 and 0.053 respectively) and to a much smaller extent in the efficiency with which cytoplasmic ATP equivalents are generated/MJ of metabolizable energy (cv 0.01). It is concluded that the current understanding of biochemical energy transduction is sufficient to permit only a crude estimate of the energy equivalents of cytoplasmic ATP but that these equivalents vary by less than 5% between both different food proteins and different food fats.(ABSTRACT TRUNCATED AT 400 WORDS)

Tissue temperatures and whole-animal oxygen consumption after exercise.

Tissue temperatures and whole-animal oxygen consumption after exercise.

Oxidative phosphorylation and adenosine triphosphatase activity in the liver mitochondria of rats administered with phenylhydrazine and hydrolyzed glucose cycloacetoacetate.

The effect of phenylhydrazine and hydroiyzed product of glucose cycloacetoacetate (GCAh) administration on the activity of adenosine triphosphatase (ATPase) in rat-liver mitochondria has been investigated. The results are discussed in relation to the efficiency of mitochondrial oxidative phosphorylation. Phenylhydrazine was found to increase the ATPase activity both in vitro and in vivo. However, preincubation or treatment with hydrolyzed glucose cycloacetoacetate resulted in an appreciable depression of this phenylhydrazine-enhanced enzyme activity. The liver mitochondria from phenylhydrazine-administered rats showed very little difference in milligrams of total protein, but the homogenates had a high protein content as compared to the preparations from normal rats and rats administered with hydrolyzed glucose cycloacetoacetate. With citrate as the substrate, normal rat-liver mitochondria exhibited a P/O ratio of 3.0. With the same substrate, the liver mitochondria from phenylhydrazine-administered rats lowered the oxygen uptake and ATP formation, thereby resulting in a decreased P/O ratio of 2.4, whereas administration of hydrolyzed glucose cycloacetoacetate prior to phenylhydrazine resulted in a partial restoration in oxygen uptake and ATP formation, and thus yielded a P/O ratio of 2.8.