Effect of the rate of oxygen consumption on muscle respiration

Summary An important role of myoglobin in red muscle is to facilitate the diffusion of oxygen for metabolism. We consider a model for muscle respiration in which the oxygen consumption is of a MichaelisMenten form. The resulting mathematical model is solved in two different ways with two different boundary conditions. The first uses the singular perturbation method of Murray (1974), while the second, which gives another justication of the simpler procedure, is a direct numerical computation of the full problem.The oxygen tension and saturation are often small. For realistic values of the Michaelis-Menten constant the oxygen tension, the saturation and the radius of the region in which the oxygen tension is negligibly small can be calculated using the constant consumption model of Murray (1974), with corrected boundary conditions (those for a Stefan problem), which in certain circumstances markedly affect the solution.B. A. T. would like to thank the Science Research Council of the United Kingdom for their financial support.     

Biphasic oxygen kinetics of cellular respiration and linear oxygen dependence of antimycin A inhibited oxygen consumption

Oxygen kinetics in fibroblasts was biphasic. This was quantitatively explained by a major mitochondrial hyperbolic component in the low-oxygen range and a linear increase of rotenone-and antimycin A-inhibited oxygen consumption in the high-oxygen range. This suggests an increased production of reactive oxygen species and oxidative stress at elevated, air-level oxygen concentrations. The high oxygen affinity of mitochondrial respiration provides the basis for the maintenance of a high aerobic scope at physiological low-oxygen levels, whereas further pronounced depression of oxygen pressure induces energetic stress under hypoxia.     

Annual cycle of oxygen consumption (QO 2) in a hibernating animalthe garden dormouse: reduction of the QO 2 and hypothermia following fasting]

A circannual cycle of oxygen consumption (QO2) was observed in active garden dormice maintained during 19 months under external conditions for temperature and lighting. QO2 was minimum in october and maximum in may. In the same animals deprivation of food induced hypothermia and a 23% decrease in QO2 which always preceeded hypothermia. No circannual rhythm was noted in the amplitude of this reduction.     

Lack of proportionality between gene dosage and total muscle protein content in the rat heart.

Counting of isolated cardiomyocytes has demonstrated that their number was 16.8 +/- 0.6 10(6) in both ventricles of weanling rats (28 days after birth), growing in litters of four (fast-growing). In rats growing in litters of 16 (slow-growing), the myocyte number was 11.8 +/- 0.8 10(6). In the control group (8 sucklings per litter), there were 14.2 +/- 10(6) cardiomyocytes. The fast-growing rats had more octoploid cells than slow-growing ones. Considering ploidy and cell number, the total number of myocyte genomes in fast-growing animals was 45% higher than in slow-growing ones. The total content of contractile proteins in fast-growing weanling animals was higher by 28% while sarcoplasmic proteins were 8% higher. This lack of correspondence between the number of myocyte genomes and muscle protein content was even more pronounced at the age of 110 days. The results are compared with the cytophotometric data concerning the lack of correspondence between the total protein content in a myocyte and its DNA amount and chromosome number, i.e., total dosage of the myocyte genes.     

Similar mitochondrial activation kinetics in wild-type and creatine kinase-deficient fast-twitch muscle indicate significant Pi control of respiration

Past simulations of oxidative ATP metabolism in skeletal muscle have predicted that elimination of the creatine kinase (CK) reaction should result in dramatically faster oxygen consumption dynamics during transitions in ATP turnover rate. This hypothesis was investigated. Oxygen consumption of fast-twitch (FT) muscle isolated from wild-type (WT) and transgenic mice deficient in the myoplasmic (M) and mitochondrial (Mi) CK isoforms (MiM CK(-/-)) were measured at 20°C at rest and during electrical stimulation. MiM CK(-/-) muscle oxygen consumption activation kinetics during a step change in contraction rate were 30% faster than WT (time constant 53 ± 3 vs. 69 ± 4 s, respectively; mean ± SE, n = 8 and 6, respectively). MiM CK(-/-) muscle oxygen consumption deactivation kinetics were 380% faster than WT (time constant 74 ± 4 s vs. 264 ± 4 s, respectively). Next, the experiments were simulated using a computational model of the oxidative ATP metabolic network in FT muscle featuring ADP and Pi feedback control of mitochondrial respiration (J. A. L. Jeneson, J. P. Schmitz, N. A. van den Broek, N. A. van Riel, P. A. Hilbers, K. Nicolay, J. J. Prompers. Am J Physiol Endocrinol Metab 297: E774-E784, 2009) that was reparameterized for 20°C. Elimination of Pi control via clamping of the mitochondrial Pi concentration at 10 mM reproduced past simulation results of dramatically faster kinetics in CK(-/-) muscle, while inclusion of Pi control qualitatively explained the experimental observations. On this basis, it was concluded that previous studies of the CK-deficient FT muscle phenotype underestimated the contribution of Pi to mitochondrial respiratory control.     

Diffusion and consumption of oxygen in the resting frog sartorius muscle

Adaptations of the method of Takahashi et al. (1966. J. Gen. Physiol. 50:317-333) were used to test the validity of the one-dimensional diffusion equation for O2 in the resting excised frog sartorius muscle. This equation is: (formula: see text) where x is the distance perpendicular to the muscle surface. t is time, P(x, t) is the partial pressure of O2,D and alpha are the diffusion coefficient and solubility for O2 in the tissue, and Q is the rate of O2 consumption. P(O, t), the time-course of PO2 at one muscle surface, was measured by a micro-oxygen electrode. Transients in the PO2 profile of the muscle were induced by two methods: (a) after an equilibration period, one surface was sealed off by a disc in which the O2 electrode was embedded; (b) when PO2 at this surface reached a steady state, a step change was made in the PO2 at the other surface. With either method, the agreement between the measured P(O, t) and that predicted by the diffusion equation was excellent, making possible the calculation of D and Q. These two methods yielded statistically indistinguishable results, with the following pooled means (+/- SEM): (formula: see text) At each temperature, D was independent of muscle thickness (range, 0.67-1.34 mm). The activation energy (EA) for diffusion of oxygen in muscle was -3.85 kcal/mol, which closely matches the corresponding value in water. Together with absolute values of D in water taken from the literature, the present data imply that (Dmuscle/DH2O) is in the range 0.59-0.69. This value, and that of EA, are in agreement with the theory of Wang (1954, J. Am. Chem. Soc. 76:4755-4763), suggesting that with respects to the diffusion of O2, to a useful approximation, frog skeletal muscle may be considered simply as a homogeneous protein solution.     

Electromechanical coupling between skeletal and cardiac muscle. Implications for infarct repair

Skeletal myoblasts form grafts of mature muscle in injured hearts, and these grafts contract when exogenously stimulated. It is not known, however, whether cardiac muscle can form electromechanical junctions with skeletal muscle and induce its synchronous contraction. Here, we report that undifferentiated rat skeletal myoblasts expressed N-cadherin and connexin43, major adhesion and gap junction proteins of the intercalated disk, yet both proteins were markedly downregulated after differentiation into myo-tubes. Similarly, differentiated skeletal muscle grafts in injured hearts had no detectable N-cadherin or connexin43; hence, electromechanical coupling did not occur after in vivo grafting. In contrast, when neonatal or adult cardiomyocytes were cocultured with skeletal muscle, approximately 10% of the skeletal myotubes contracted in synchrony with adjacent cardiomyocytes. Isoproterenol increased myotube contraction rates by 25% in coculture without affecting myotubes in monoculture, indicating the cardiomyocytes were the pacemakers. The gap junction inhibitor heptanol aborted myotube contractions but left spontaneous contractions of individual cardiomyocytes intact, suggesting myotubes were activated via gap junctions. Confocal microscopy revealed the expression of cadherin and connexin43 at junctions between myotubes and neonatal or adult cardiomyocytes in vitro. After microinjection, myotubes transferred dye to neonatal cardiomyocytes via gap junctions. Calcium imaging revealed synchronous calcium transients in cardiomyocytes and myotubes. Thus, cardiomyocytes can form electromechanical junctions with some skeletal myotubes in coculture and induce their synchronous contraction via gap junctions. Although the mechanism remains to be determined, if similar junctions could be induced in vivo, they might be sufficient to make skeletal muscle grafts beat synchronously with host myocardium.     

Adenylate control of respiration in plants: the contribution of rotenone-insensitive electron transport to ADP-limited oxygen consumption by soybean mitochondria

The aim was to test the hypothesis that rotenone-insensive electron transport (bypass of complex I) may underlie rapid state 4 (ADP-limited) mitochondrial respiration. A comparison of mitochondria from soybean ( Glycine max L. cv. Bragg) cotyledons and nodules showed that ADP-sufficient (state 3) malate plus pyruvate oxidation by mitochondria from 7-day-old cotyledons was inhibited 50% by rotenone and state 4 rates were rapid, whereas nodule mitochondria were 80% inhibited by rotenone and had slower state 4 rates of malate plus pyruvate oxidation. Respiration of malate alone (pH 7.6) by cotyledon mitochondria was slow, especially in the absence of ADP; subsequent addition of pyruvate dramatically increased state 4 oxygen uptake concomitant with a rapid rise in mitochondrial NADH (determined by fluorimetry). Rotenone had no effect on this increased rate of state 4 respiration. The rate of malate oxidation by nodule mitochondria was relatively rapid compared with cotyledon mitochondria. The addition of pyruvate in state 4 caused a slow increase in matrix NADH and only a slight stimulation of oxygen uptake. Rotenone inhibited state 4 malate plus pyruvate oxidation by 50% in these mitochondria. From a large number of cotyledon and nodule mitochondrial preparations, a close correlation was found between the rate of state 4 oxygen uptake and rotenone-resistance. During cotyledon development increased rotenone-resistance was associated with an increase in the alternative oxidase. Addition of pyruvate to cotyledon mitochondria, during state 4 oxidation of malate in the presence of antimycin A, significantly stimulated O₂ uptake and also almost eliminated respiratory control. Such combined operation of the rotenone-insensitive bypass and the alternative oxidase in vivo will significantly affect the extent to which adenylates control the rate of electron transport.     

Relationship between pulmonary O2 uptake kinetics and muscle deoxygenation during moderate-intensity exercise.

The temporal relationship between the kinetics of phase 2 pulmonary O2 uptake (Vo -->Vo2p) and deoxygenation of the vastus lateralis muscle was examined during moderate-intensity leg-cycling exercise. Young adults (5 men, 6 women; 23 +/- 3 yr; mean +/- SD) performed repeated transitions on 3 separate days from 20 W to a constant work rate corresponding to 80% of lactate threshold. Breath-by-breath Vo2p was measured by mass spectrometer and volume turbine. Deoxyhemoglobin (HHb), oxyhemoglobin, and total hemoglobin and myoglobin were sampled each second by near-infrared spectroscopy (Hamamatsu NIRO-300). Vo2p data were filtered, interpolated to 1 s, and averaged to 5-s bins; HHb data were averaged to 5-s bins. Phase 2 Vo2p data were fit with a monoexponential model. For HHb, a time delay (TDHHb) from exercise onset to an increase in HHb was determined, and thereafter data were fit with a monoexponential model. The time constant for Vo2p (30 +/- 8 s) was slower (P < 0.01) than that for HHb (10 +/- 3 s). The TDHHb before an increase in HHb was 13 +/- 2 s. The possible mechanisms of the TDHHb are discussed with reference to metabolic activation and matching of local muscle O2 delivery and O2 utilization. After this initial TDHHb, the kinetics of local muscle deoxygenation were faster than those of phase 2 Vo2p (and presumably muscle O2 consumption), reflecting increased O2 extraction and a mismatch between local muscle O2 consumption and perfusion.     

Reappraisal of diffusion, solubility, and consumption of oxygen in frog skeletal muscle, with applications to muscle energy balance

Previously we tested the validity of the one-dimensional diffusion equation for O2 in the excised frog sartorius muscle and used it to measure the diffusion coefficient (D) for O2 in this muscle and the time course of its rate of O2 consumption (Qo2) after a tetanus (Mahler, 1978, 1979, J. Gen. Physiol., 71:533-557, 559-580, 73:159-174). A transverse section of the frog sartorius is in fact well fit by a hemi-ellipse with width divided by maximum thickness averaging 5.1 +/- 0.2. Using the previous techniques with the two-dimensional diffusion equation and this hemi-elliptical boundary yields a value for D that is 30% smaller than reported previously; the revised values at 0, 10, and 22.8 degrees C are 6.2, 7.9, and 10.8 X 10(-6) cm2/s, respectively. After a tetanus at 20 degrees C, Qo2 rose quickly to a peak and then declined exponentially, with a time constant (tau) approximately 15% faster than that reported previously; tau averaged 2.1 min in Rana temporaria and 2.6 min in Rana pipiens. A technique was devised to measure the solubility (alpha) of O2 in intact, respiring muscles, and yielded alpha (muscle)/alpha (H2O) = 1.26 +/- 0.04. With these modifications, the values for O2 consumption obtained with the diffusion method were in agreement with those measured by the direct method of Kushmerick and Paul (1976, J. Physiol. [Lond.]., 254:693-709). Using results from both methods, at 20 degrees C the ratio of phosphorylcreatine split during a tetanus to O2 consumption during recovery ranged from 5.2 to 6.2 mumol/mumol, and postcontractile ATP hydrolysis was estimated to be 13.6 +/- 4.1 (n = 3) nmol/mumol total creatine.     

Determination of the respiration kinetics for mycelial pellets of Phanerochaete chrysosporium.

In mycelial pellet cultures of the white rot basidiomycete Phanerochaete chrysosporium, low oxygen concentration negatively affects the production of the extracellular lignin peroxidases and manganese peroxidases which are key components of the lignin-degrading system of this organism. To test the hypothesis that oxygen limitation in the pellets is responsible for this effect, oxygen microelectrodes were used to determine oxygen concentration gradients within the mycelial pellets of P. chrysosporium. Pellets were removed from oxygenated cultures, allowed to equilibrate with air, and probed with oxygen microelectrodes. The oxygen profiles were modelled assuming that O2 uptake follows a Michaelis-Menten relationship. The Vmax and Km values for oxygen uptake were 0.76 +/- 0.10 g/m3 of pellet per s and 0.5 +/- 0.3 g/m3, respectively. These kinetic values were used to predict respiration rates in air-flushed cultures, oxygen-flushed cultures, and cultures with large pellets (diameter greater than 6 mm). The predicted respiration rates were independently validated by experimentally measuring the evolution of carbon dioxide from whole cultures.     

Determination of the respiration kinetics for mycelial pellets of Phanerochaete chrysosporium.

In mycelial pellet cultures of the white rot basidiomycete Phanerochaete chrysosporium, low oxygen concentration negatively affects the production of the extracellular lignin peroxidases and manganese peroxidases which are key components of the lignin-degrading system of this organism. To test the hypothesis that oxygen limitation in the pellets is responsible for this effect, oxygen microelectrodes were used to determine oxygen concentration gradients within the mycelial pellets of P. chrysosporium. Pellets were removed from oxygenated cultures, allowed to equilibrate with air, and probed with oxygen microelectrodes. The oxygen profiles were modelled assuming that O2 uptake follows a Michaelis-Menten relationship. The Vmax and Km values for oxygen uptake were 0.76 +/- 0.10 g/m3 of pellet per s and 0.5 +/- 0.3 g/m3, respectively. These kinetic values were used to predict respiration rates in air-flushed cultures, oxygen-flushed cultures, and cultures with large pellets (diameter greater than 6 mm). The predicted respiration rates were independently validated by experimentally measuring the evolution of carbon dioxide from whole cultures.     

The relationship between energy-dependent phagocytosis and the rate of oxygen consumption in Tetrahymena.

The induction of high rates of food vacuole formation in Tetrahymena pyriformis increased the rate of respiration in exponentially growing cells by 17% and in starving cells by 47.5%. The increased rate of oxygen uptake was caused by phagocytosis itself, as shown by comparing the rates of respiration of a Tetrahymena mutant exposed to particles at the permissive or restrictive temperatures for food vacuole formation. During cell division, heat-synchronized cells in rich, particle-supplemented medium showed a significant decrease in the rate of respiration. Furthermore, dimethyl sulphoxide, in concentrations sufficient to block food vacuole formation, suppressed the rate of respiration to a level similar to that of starved cells. Cytochalasin B, fowever, did not reduce the rate of oxygen uptake despite the inability of the cells to complete the formation of food vacuoles during treatment; a possible explanation for this finding is discussed. There was a strong correlation between formation of food vacuoles and a high metabolic rate in Tetrahymena.     

Variation in the link between oxygen consumption and ATP production,and its relevance for animal performance

It is often assumed that an animal''s metabolic rate can be estimated through measuring the whole-organism oxygen consumption rate. However, oxygen consumption alone is unlikely to be a sufficient marker of energy metabolism in many situations. This is due to the inherent variability in the link between oxidation and phosphorylation; that is, the amount of adenosine triphosphate (ATP) generated per molecule of oxygen consumed by mitochondria (P/O ratio). In this article, we describe how the P/O ratio can vary within and among individuals, and in response to a number of environmental parameters, including diet and temperature. As the P/O ratio affects the efficiency of cellular energy production, its variability may have significant consequences for animal performance, such as growth rate and reproductive output. We explore the adaptive significance of such variability and hypothesize that while a reduction in the P/O ratio is energetically costly, it may be associated with advantages in terms of somatic maintenance through reduced production of reactive oxygen species. Finally, we discuss how considering variation in mitochondrial efficiency, together with whole-organism oxygen consumption, can permit a better understanding of the relationship between energy metabolism and life history for studies in evolutionary ecology.     

Evidence for an inverse relationship between the ventilatory response to exercise and the maximum whole body oxygen consumption value

Ventilatory responses to submaximal exercise loads indicate that in a population of 895 physically active and sedentary male and female subjects, exercise ventilation is inversely related to predicted VO2max. The correlation coefficients for males and females in this relationship are 0.61 (P less than 0.0001) and 0.26 (P less than 0.0001) respectively. The slopes of regression lines for VE/VO2 and VO2max in female and male subjects are -2.59 and -0.91 respectively. This is associated with changes in composition of the expired air in that PCO2 increases and PO2 decreases with greater VO2max. The difference between the mean oxygen and carbon dioxide partial pressures in expired air of individuals in the highest and lowest VO2max ranges are 1.2 kPa (9 mm Hg) and 0.8 kPa (6 mm Hg) respectively.     

The regulation index: a new method for assessing the relationship between oxygen consumption and environmental oxygen

Critical oxygen pressure (P(C)) is used in respiratory physiology to measure the response to hypoxia. P(C) defines the partial pressure of oxygen (Po(2)) at which an oxygen regulator switches to a conformer. However, not all animals show such clear patterns in oxygen consumption rate (Mo2), and there are many methods for determining P(C). This study assesses two methods that determine regulatory ability and four that calculate P(C). A new method, the regulation index (RI), assigns to an animal a relative measure of regulatory ability by calculating the area under the Mo2 versus Po(2) curve that is greater than a linear trend. The six methods are applied to developmental Mo2 data of two amphibians, Pseudophryne bibronii and Crinia georgiana. The four methods used to determine P(C) produced similar results but failed to identify the increase in regulation on hatching in C. georgiana or the greater regulation in larval C. georgiana compared with P. bibronii. Of the two methods that evaluated regulation, only the RI satisfactorily represented the entire range of Po(2). The RI is advantageous because it has clearly defined limits and does not constrain data to fit any single pattern. The RI can be used in concert with P(C), which can be easily calculated during the RI analysis, to provide a clearer definition of the Mo2 response to environmental Po(2).