International Congress Series 1275 (2004) 22 31

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Oxidative fuel selection: adjusting mix and flux to stay alive

Jean-Michel Weber*, Franc ois Haman
Biology Department, University of Ottawa, 30 Marie Curie, Ottawa, Ontario, Canada K1N 6N5

Abstract. To be able to match ATP supply with demand, animals must ensure adequate delivery of metabolic fuels and oxygen to tissue mitochondria. Therefore, the mixture of fuels provided and their individual flux must be tightly orchestrated to cope with changing physiological needs. In exercising mammals, metabolic rateexpressed relatively to the aerobic maximum: %VO2 maxdetermines what mixture of oxidative fuels is being used. This simple model of fuel selection accurately predicts the relative contributions of lipids and carbohydrates to total metabolism, and it applies widely across body sizes, aerobic capacities, and even to exercise in hypoxic environments. However, it is also becoming obvious that significant exceptions to this pattern exist in other vertebrates that rely more heavily on lipids (e.g., migrating birds) or proteins (e.g., migrating salmonids), or for stresses other than exercise (e.g., cold exposure in mammals). Instantaneous fuel use is determined by multiple interacting mechanisms involving fuel availability, storage location, muscle recruitment, fiber recruitment within each muscle, and metabolic pathway selection within each fiber. These various mechanisms are being characterized in more detail to try designing a general model of fuel selection applicable to a wider range of animals and physiological stresses. D 2004 Elsevier B.V. All rights reserved.
Keywords: Animal energetics; Energy metabolism; Metabolic substrate; Exercise; Shivering thermogenesis; Migration; Hibernation; Metabolic depression; Lipid; Carbohydrate; Protein

1. Introduction The ability to adjust energy expenditure to cope with changing physiological circumstances is a key feature of organismal survival, and a lot of research has focused on understanding the fundamental mechanisms involved in the upregulation
* Corresponding author. Tel.: +1 613 562 5800 6007; fax: +1 613 562 5486. E-mail address: jmweber@science.uottawa.ca (J.-M. Weber). 0531-5131/ D 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.ics.2004.09.043

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(exercise, cold exposure, lactation) or depression of metabolism (fasting, hypoxia, torpor, hibernation, estivation). However, merely changing total flux of O2 and substrates to mitochondria is not sufficient to ensure long-term survival because internal fuel sources are extremely diverse in size, chemical properties, and storage locations. Therefore, the capacity to select an adequate mixture of metabolic fuels (change in mix) and to modulate this blend (change in flux) is another essential requirement for survival. This paper examines the strategies used by animals to alter their pattern of fuel selection together with the supply rate of each individual substrate to mitochondria. The tight regulation of mix and flux is necessary to balance rates of ATP production with prevailing rates of ATP utilization. Locomotion, thermogenesis, and metabolic depression are perhaps the most striking examples of functional needs that critically depend on modulating the quality and quantity of oxidative fuel supply. 2. Oxidative fuel diversity To produce ATP for long-term activities, animals must rely on the oxidation of lipids, carbohydrates, and proteins stored within their tissues. These metabolic fuel reserves are obtained from the diet and are usually replenished during periods of rest, recovery from exercise, or seasonal preparation for prolonged fasting associated with long-distance migration or hibernation. The oxidation of each fuel presents clear advantages and disadvantages for any particular physiological situation. To illustrate the convenience and constraints afforded by such diversity, different criteria can be used for comparing the various sources of energy, and, in this context, key characteristics of the fuels available are summarized in Table 1. Lipids represent the most concentrated source of energy in living organisms for two important reasons: (i) they are the most chemically reduced of all fuels and (ii) they can be stored without water. Therefore, animals favour lipids for energy storage, and most land species could simply not afford to transport the additional weight associated with alternative fuels. Despite their enormous bweight handicapQ, carbohydrates are essential when ATP must be produced at high rates, without delay, or, possibly, when O2 availability is compromised. For many aquatic animals, weight is not an issue because fuel reserves do not have to be carried against gravity. Unlike lipids, carbohydrates and

Table 1 Comparison of the different oxidative fuels available for ATP synthesis Unit Isocaloric weight Percent total energy reserves Maximal rate of ATP production Time to reach maximal rates Energy per volume O2 g fuel MJ1 % Amol ATP g1 min1 min 1 kJ l O 2 Lipids 26 85 20 N30 19.8 Carbohydrates 239 1 30 b2 21.1 Proteins 55 14

18.7

Adapted from Refs. [14]. Lipid values are based on triacylglycerol with an average mammalian fatty acid composition. Carbohydrate values were calculated for natural glycogen with an average level of hydration. For proteins, values were calculated for ureotelic animals and only for the mobilizable fraction of total proteins.

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proteins are soluble in aqueous biological fluids and do not depend on carrier molecules like serum albumin and fatty acid-binding proteins (FABP) for circulatory and cytoplasmic transport. The complete oxidation of all fuels produces CO2 and H2O, two end-products that can usually be managed without problems. However, protein oxidation presents a unique metabolic limitation because it also yields noxious ammonia that must be eliminated or detoxified. The size of fuel reserves may be altered drastically in preparation for specific physiological challenges. As an extreme example, lipid stores can be increased to reach up to 50% of total body mass before hibernation in small mammals [5] or longdistance migration in birds [6]. To a lesser extent, fuel storage is also affected by endurance training [7,8] or large shifts in diet composition [9]. In turn, these changes in internal substrate availability can influence the pattern of fuel selection during exercise, cold exposure, or metabolic depression. In addition to inflating or decreasing the size of specific energy reserves, animals can also change the distribution of each type of fuel among storage sites. During sustained exercise, ATP production of locomotory muscles depends on the oxidation of both, intramuscular fuels (muscle glycogen and muscle triacylglycerol), and circulatory fuels brought to working muscles from remote storage sites (hepatic glucose and adipose tissue lipids) [10]. To be able to reach high rates of oxidative fuel supply to muscle mitochondria, very aerobic mammals (high VO2 max) rely relatively more on intramuscular fuels, and relatively less on circulatory fuels than sedentary species (low VO2 max) [11,12]. This strategy is necessary to circumvent significant constraints associated with the multiple trans -membrane crossings required to bring fuels from distant storage locations [1315]. Nowhere is this adaptation made more obvious than in the way intramuscular lipid reserves are organized: all muscle lipid droplets are actually in direct contact with mitochondria [16,17]. With such a spatial arrangement, lipid transport from storage to the enzymatic

Fig. 1. (A) Transmission electron micrograph of dog triceps muscle illustrating the close association between intramuscular lipid droplets loaded with triacylglycerol (li) and mitochondria (mt). Adapted from Ref. [17]. (B) Fuel selection pattern for mammalian exercise. Changes in the relative contribution of carbohydrate oxidation (CHO; open symbols) and lipid oxidation (closed symbols) to total oxygen consumption of the whole organism (VO2) as a function of exercise intensity (%VO2 max). Values are for dogs (o, ) [18], goats (q, z) [18], humans (5, n) [19] and rats (D, E) [20,21]. Results include values for trained and untrained rats acclimated to normoxia or to hypoxia.

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machinery of energy metabolism is reduced to its simplest form: one single membrane to cross (see Fig. 1A). 3. Exercising mammals: a robust model of fuel selection Almost all the information available on fuel selection comes from one experimental model: mammalian exercise . For this group of animals, the observed pattern is surprisingly simple. Total ATP production can be attributed exclusively to lipid and carbohydrate oxidation because the contribution from proteins is minimal [17,22]. More importantly, exercise intensity, expressed in relation to the aerobic maximum (%VO2 max), determines the relative importance of lipids and carbohydrates according to the relationship presented in Fig. 1B. The contribution of carbohydrates increases progressively, and that of lipids decreases progressively as exercise intensifies. Therefore, the oxidation of each one of these two fuels is responsible for half the metabolic rate of the whole organism at a work intensity of about 50% VO2 max (or the work intensity sometimes referred to as the bcrossover pointQ; Ref. [22]). Different approaches have been used to build this model and to ensure that it could be generalized to all mammals. Because the balance between lipids and carbohydrates originally seemed to depend on %VO2 max, the robustness of the model was assessed by exploiting various ways to manipulate aerobic capacity. Large adaptive differences in VO2 max (i.e., genetic differences) exist in nature between very sedentary and highly aerobic species. Dogs and goats of the same size were used in this context because their aerobic capacities differ by more than twofold [18]. Although highly aerobic dogs (geared for endurance exercise) were anticipated to favor the use of the ample lipid reserves available to all animals (see Table 1), results show that both species oxidize the same mixture of fuels when they exercise at the same relative intensity (Fig. 1B) [18]. Because massspecific VO2 max varies greatly with body mass, measurements were extended to smaller species (0.3 kg rats) and larger ones (70 kg humans), but without finding significant deviation from the dog-goat pattern (Fig. 1B). Finally, experiments were carried out under low oxygen availability because aerobic capacity is reduced by acclimation to hypoxia. In addition, there is a convincing theoretical reason to think that animals should favor carbohydrates when exercising in hypoxia because this fuel yields 11% more ATP per unit volume of oxygen than lipids (Table 1, but see Ref. [23] for arguments supporting an even greater difference). Again, results show that, individuals acclimated to hypoxia, running under normoxic or hypoxic conditions, follow the same pattern of fuel selection previously observed in all other mammals (Table 1) [20,24,25]. Therefore, the theoretical O2-saving advantage provided by carbohydrates seems to be outweighed by the potential danger of depleting this small, but critical energy reserve [25]. Although the relative partitioning between lipids and carbohydrates is the same for different mammals, it is important to realize that, for each relative exercise intensity, absolute rates of lipid and carbohydrate oxidation are scaled directly with VO2 max (i.e., they are more than two times higher in dogs than in goats). From these observations, we can conclude that the fuel selection model proposed for exercising mammals (Fig. 1B) is extremely robust because it is independent of aerobic capacity when tested for: adaptive variation (dog vs. goat),

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allometric variation (0.3 kg rat vs. 70 kg human), and environmental variation in O2 availability (normoxia vs. hypoxia). 4. Alternative fuel selection patterns: swimming fish, flying birds, and shivering humans Multiplying measurements in more examples of exercising mammals is unlikely to yield further useful insights. Instead, developing a theoretical framework explaining the reasons for the observed pattern appear more promising. To achieve this goal, two interrelated strategies come to mind: uncover clear exceptions to this seemingly general pattern (this section) and characterize the mechanisms available for altering fuel selection (next section). In the last few years, several examples of divergent patterns have emerged, and these exceptions could prove very useful for future research. As more information is accumulating for various animals, it is obvious that the mammalian pattern is far from universal; major differences in fuel metabolism can be found, even among vertebrates. In fish, ignoring the contribution from proteins, as it was done for mammals, could lead to enormous errors because some species are probably able to rely almost exclusively on this source of energy for sustained swimming (e.g., during the late stages of migration in sockeye salmon [3]). In addition, the three to fourfold increments in glucose flux (rate of hepatic glucose production) and fatty acid flux (rates of lypolysis and fatty acid supply) classically reported for all exercising mammals, are completely absent in rainbow trout, even during prolonged swimming [26,27]. Salmonids clearly fail to follow the mammalian model, but more research is needed to determine whether their fuel selection pattern is typical of teleosts in general. Long-distance migrant birds are another example at variance with mammals because their relative use of lipids is much higher than that predicted in Fig. 1B. They have been able to push conventional energy metabolism well beyond the limits set by the best mammalian athletes. Many bird species migrate at 1015 times their basal metabolic rate, or twice the VO2 max of same-size mammals [28]. More importantly, most of the energy used to power long-distance flights is provided by the circulation from adipose lipid reserves [29]. These characteristics are incompatible with the mammalian model, stipulating that, at intensities approaching VO2 max, over 80% of the energy comes from carbohydrates (mainly muscle glycogen) [15], and that the oxidation of circulating lipids accounts maximally for 1020% of VO2 [11,12]. Although limited quantitative information is available on the fuel metabolism of migrant birds, we can deduce from first principles that using glycogen at such high rates is impossible; glycogen reserves of the necessary magnitude do not exist in nature because their weight would prevent movement (see Table 1). During migration flights, rates of circulatory lipid oxidation are therefore at least 10, possibly 20 times higher than the maximal rates ever measured in exercising mammals. Therefore, we can conclude that the mammalian crossover curves presented in Fig. 1B must be strongly shifted to the right for long-distance migrant birds. Artificially modifying the size of energy reserves can also lead to significant adjustments in substrate use. In humans [3032] and sled dogs [8], important changes in fuel selection have been elicited by dietary manipulations (high-fat diet or glycogen loading). Therefore, bon-boardQ availability of each fuel type influences the mixture of

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metabolic substrates oxidized. Although the exact signals relaying information about the size of energy stores are still poorly understood, major advances have been made in this area. For example, leptin levels signal the size of lipid stores [33], and, on its own, this hormone has significant effects on fuel selection [34,35]. Finally, recent experiments on the effects of cold exposure in humans suggest that the fuel selection patterns of shivering and exercise are different [36]. Detailed measurements of fuel oxidation for thermogenesis show that carbohydrates play a much more important role during shivering than exercise, when these activities are compared at the same metabolic rate [32,3638]. Therefore, the fuel selection pattern of shivering humans can probably be obtained by shifting the exercise curves of Fig. 1B to the left. The fact that the same muscles use different mixtures of fuels during shivering and exercise when they function at the same metabolic rate is very intriguing. The quest for an explanation of this fascinating difference could provide novel insights on the fundamental mechanisms of fuel selection. 5. Mechanisms for selection Fuel selection can occur by changing the supply rate or the utilization rate of the different substrates available. Most of the well-characterized mechanisms of selection operate directly at the level of fuel utilization , but a simple (and often overlooked) mechanism acting at the level of fuel supply can also play an important role. Experiments on thoroughbred horses have allowed showing that the supply of circulatory fuels to working muscles is regulated in two ways [39,40]. First, the animal can change the supply rate of all the fuels provided through the circulation by adjusting cardiac output and blood flow to target tissues (coarse control of substrate flux acting indiscriminately on all circulatory fuels). Second, the supply rate of individual fuels can be modulated separately by changing their concentration in the blood (fine control of flux acting specifically on each fuel). Together, these two mechanisms allow adjusting the flux of each blood-borne substrate, thereby setting a fuel mixture adequate for present conditions. The last part of this review deals with the best-characterized selection mechanisms that act at the level of fuel utilization in skeletal muscle. These mechanisms can be divided in three categories based on the level of organization where they exert their effects. Changing the mixture of fuels can be done by selective recruitment of: (i) different muscles, (ii) different fibers within the same muscle, or (iii) different metabolic pathways within the same fiber. The selective recruitment of different muscles was first demonstrated in fish where red muscle (made of slow fibers specialized for lipid oxidation) and white muscle (made of fast fibers specialized for carbohydrate oxidation) are spatially separated (e.g., see Refs. [41,42]). The same mechanism can also regulate fuel selection in exercising mammals and birds, although their muscles are made of mixed fibers. Using blood flow as an index of recruitment, it has been possible to show that muscles with predominantly slow fibers (specialized for fat oxidation) are already active at low work intensities, whereas muscles with predominantly fast fibers (oxidizing carbohydrates) are only recruited at high exercise intensities [43,44]. Similarly, it has been accepted for a long time that fuel metabolism can be regulated within individual muscles through the selective recruitment of different fiber populations specialized for different substrates [18]. Direct proof of this mechanism has only been

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provided very recently by making simultaneous measurements of fiber recruitment and fuel utilization. Clear enough electromyographic (EMG) signals cannot be obtained during exercise because of the large background noise created by limb movements. Therefore, the problem was eliminated by using an alternative experimental model: shivering muscles of humans exposed to cold. During high-intensity shivering, large differences in fuel selection between individuals (i.e., carbohydrates accounting for 33% to 78% of metabolic rate) are explained by differences in the recruitment of fast (type II) fibers, specialized for carbohydrate oxidation [36]. Interestingly, the alternative mechanism of selectionthe recruitment of different metabolic pathways within the same muscle fibersis used during low-intensity shivering. Detailed EMG analyses reveal that glycogen-depleted and glycogen-loaded individuals can have the same (low) thermogenic rate, but by using widely different fuel mixtures within the same (type I) muscle fibers [32,38]. Biochemical mechanisms of fuel selection have been investigated in mammalian muscles and they continue to be the subject of intense research [19]. Numerous extra- and intracellular signal molecules have been implicated. Since the early sixties, the bglucosefatty acid cycleQ of Randle et al. [45] has often been invoked to explain how the balance between lipids and carbohydrates can be achieved. When fatty acid availability is high, carbohydrate oxidation is reduced and lipid oxidation is stimulated. Randle et al. [45] proposed that high plasma fatty acid concentration caused these changes by suppressing the activation of the pyruvate dehydrogenase complex (PDC; through a rise in the mitochondrial acetyl-CoA/CoA ratio) and by decreasing glycolysis (through inhibition of phosphofructokinase via elevated citrate levels). Although PDC is still considered a significant element in the regulation of fuel selection [46], it has become clear that other mechanisms such as the direct inhibition of glucose transporters (GLUT-4) and of glucose phosphorylation can also play important roles [47]. Over the last few years, malonyl-CoA has also attracted some attention as a possible regulator of fuel selection [48]. The high glycolytic flux associated with intense exercise causes the accumulation of acetyl-CoA, and it has been proposed that this would increase cytosolic malonyl-CoA, thereby causing the inhibition of carnitine palmitoyltransferase I (CPT I) and limiting fatty acid entry into mitochondria. However, direct measurements of malonyl-CoA levels suggest that this probable intramuscular signal does not increase during heavy exercise (at least in rats and humans). Therefore, this mechanism remains doubtful [49]. Ongoing research suggests the involvement of free carnitine levels and intracellular pH that would both inhibit CPT I when low [19,50]. Finally, it has been suggested that fatty acid-binding proteins could play a significant role in fuel selection through their direct regulation of glycolytic enzymes [51]. 6. Conclusions An array of fuels with different properties is available for energy metabolism, and each one (or each mix) is advantageous for a particular physiological situation. Fuel selection strategies are geared to manage energy reserves in a way to avoid the complete depletion of any individual fuel to preserve the capacity to respond adequately to all life challenges. Exercising mammals follow a simple pattern of fuel utilization whereby the balance between lipids and carbohydrates is determined by relative work intensity (%VO2 max).

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This model is very robust because it is independent of aerobic capacity across adaptive, allometric, and environmental variation. In contrast, swimming fish, long-distance migrant birds, and shivering humans follow different patterns of fuel selection whose study will provide important novel insights on fundamental aspects of energy metabolism. The mechanisms responsible for the regulation of fuel selection in working muscles include the selective recruitment of different muscles, of different fibers within the same muscle, and of different metabolic pathways within the same fiber. Reconciling detailed mechanistic information with the fuel selection patterns observed in the whole organism remains a major challenge for future research. Acknowledgements This research program is supported by NSERC grants (Canada) to J.-M. Weber. References
[1] J.M. Berg, J.L. Tymoczko, L. Stryer, Biochemistry, 5th ed., W.H. Freeman, New York, 2002. [2] R.W. Hill, G.A. Wyse, M. Anderson, Animal Physiology, Sinauer Associates, Sunderland, MA, 2004. [3] P.W. Hochachka, G.N. Somero, Biochemical Adaptation. Mechanism and Process in Physiological Evolution, Oxford University Press, New York, 2002. [4] K. Schmidt-Nielsen, Animal Physiology, 4th ed., Cambridge Univerity Press, New York, 1990. [5] M.M. Humphries, D.W. Thomas, D.L. Kramer, The role of energy availability in mammalian hibernation: a costbenefit approach, Physiol. Biochem. Zool. 76 (2) (2003) 165 179. [6] C.R. Blem, Avian energy storage, Curr. Ornithol. 7 (1990) 59 113. [7] B.F. Hurley, et al., Muscle triglyceride utilization during exercise: effect of training, J. Appl. Physiol. 60 (1986) 562 567. [8] A.J. Reynolds, et al., Effect of diet and training on muscle glycogen storage and utilization in sled dogs, J. Appl. Physiol. 79 (5) (1995) 1601 1607. [9] H.W. Goforth, et al., Effects of depletion exercise and light training on muscle glycogen supercompensation in men, Am. J. Physiol. 285 (6) (2003) E1304 E1311. [10] J.-M. Weber, Pathways for oxidative fuel provision to working muscles: ecological consequences of maximal supply limitations, Experientia 48 (6) (1992) 557 564. [11] J.-M. Weber, et al., Design of the oxygen and substrate pathways: III. Partitioning energy provision from carbohydrates, J. Exp. Biol. 199 (1996) 1659 1666. [12] J.-M. Weber, et al., Design of the oxygen and substrate pathways: IV. Partitioning energy provision from fatty acids, J. Exp. Biol. 199 (1996) 1667 1674. [13] J.-M. Weber, Oxidative metabolism in muscle cells, in: R.G. Crystal, J.B. West, E.R. Weibel, P.J. Barnes (Eds.), The Lung: Scientific Foundations, 2nd ed., Lippincott-Raven, Philadelphia, 1997, pp. 1883 1888, Chap. 139. [14] J.-M. Weber, Adjusting maximal fuel delivery for differences in demand, in: E.R. Weibel, C.R. Taylor, L.C. Bolis (Eds.), Principles of Animal Design: The Optimization and Symmorphosis Debate, Cambridge University Press, Cambridge, UK, 1998, pp. 249 254, Chap. 9.3. [15] E.R. Weibel, et al., Design of the oxygen and substrate pathways: VII. Different structural limits for O2 and substrate supply to muscle mitochondria, J. Exp. Biol. 199 (1996) 1699 1709. [16] R. Vock, et al., Design of the oxygen and substrate pathways: VI. Structural basis of intracellular substrate supply to mitochondria in muscle cells, J. Exp. Biol. 199 (1996) 1689 1697. [17] C.R. Taylor, et al., Design of the oxygen and substrate pathways: I. Model and strategy to test symmorphosis in a network structure, J. Exp. Biol. 199 (1996) 1643 1649. [18] T.J. Roberts, et al., Design of the oxygen and substrate pathways: II. Defining the upper limits of carbohydrate and fat oxidation, J. Exp. Biol. 199 (1996) 1650 1658.

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[19] L.J.C. van Loon, et al., The effects of increasing exercise intensity on muscle fuel utilisation in humans, J. Physiol. 536 (1) (2001) 295 304. [20] G.B. McClelland, et al., High-altitude acclimation increases the triacylglycerol/fatty acid cycle at rest and during exercise, Am. J. Physiol. 281 (2001) E537 E544. [21] G.A. Brooks, C.M. Donovan, Effect of endurance training on glucose kinetics during exercise, Am. J. Physiol. 244 (1983) E505 E512. [22] G.A. Brooks, J. Mercier, Balance of carbohydrate and lipid utilization during exercise: the bcrossoverQ concept, J. Appl. Physiol. 76 (6) (1994) 2253 2261. [23] J.F. Hutter, H.M. Piper, P.G. Spieckermann, Effect of fatty acid oxidation on efficiency of energy production in rat heart, Am. J. Physiol. 249 (1985) H723 H728. [24] G.B. McClelland, P.W. Hochachka, J.-M. Weber, Effect of high-altitude acclimation on NEFA turnover and lipid utilization during exercise in rats, Am. J. Physiol. 277 (1999) E1095 E1102. [25] G.B. McClelland, P.W. Hochachka, J.-M. Weber, Carbohydrate utilization during exercise after high-altitude acclimation: a new perspective, Proc. Natl. Acad. Sci. U. S. A. 95 (1998) 10288 10293. [26] S.F. Bernard, et al., Glycerol and fatty acid kinetics in rainbow trout: effects of endurance swimming, J. Exp. Biol. 202 (1999) 279 288. [27] D.S. Shanghavi, J.-M. Weber, Effects of sustained swimming on hepatic glucose production of rainbow trout, J. Exp. Biol. 202 (1999) 2161 2166. [28] P.J. Butler, A.J. Woakes, Fat storage and fat metabolism in relation to migration, in: E. Gwinner (Ed.), Bird Migration: Physiology and Ecophysiology, Springer-Verlag, New York, 1990, pp. 300 318. [29] L. Jenni, S. Jenni-Eiermann, Fuel supply and metabolic constraints in migrating birds, J. Avian Biol. 29 (1998) 521 528. [30] J.A. Hawley, F. Brouns, A. Jeukendrup, Strategies to enhance fat utilisation during exercise, Sports Med. 25 (4) (1998) 241 257. [31] S.D. Phinney, et al., The human metabolic response to chronic ketosis without caloric restriction: preservation of submaximal exercise capability with reduced carbohydrate oxidation, Metabolism 32 (8) (1983) 769 776. [32] F. Haman, et al., Effects of carbohydrate availability on sustained shivering: I. Oxidation of plasma glucose, muscle glycogen and proteins, J. Appl. Physiol. 96 (1) (2004) 32 40. [33] R.S. Ahima, J.S. Flier, Leptin, Annu. Rev. Physiol. 62 (2000) 413 437. [34] S.P. Reidy, J.-M. Weber, Accelerated substrate cycling: a new energy-wasting role for leptin in vivo, Am. J. Physiol. 282 (2002) E312 E317. [35] S. Reidy, J.-M. Weber, Leptin: an essential regulator of lipid metabolism, Comp. Biochem. Physiol., A 125 (3) (2000) 285 297. [36] F. Haman, S.R. Legault, J.-M. Weber, Fuel selection during intense shivering in humans: EMG pattern reflects carbohydrate oxidation, J. Physiol. 556 (1) (2004) 305 313. [37] F. Haman, et al., Effect of cold exposure on fuel utilization in humans: plasma glucose, muscle glycogen and lipids, J. Appl. Physiol. 93 (2002) 77 84. [38] F. Haman, et al., Effects of carbohydrate availability on sustained shivering: II. Relating muscle recruitment to fuel selection, J. Appl. Physiol. 96 (1) (2004) 41 49. [39] J.-M. Weber, et al., Cardiac output and oxygen consumption in exercising thoroughbred horses, Am. J. Physiol. 253 (1987) R890 R895. [40] J.-M. Weber, et al., Lactate kinetics in exercising thoroughbred horses: regulation of turnover rate in plasma, Am. J. Physiol. 253 (1987) R896 R903. [41] B.C. Jane, G.V. Lauder, How swimming fish use slow and fast muscle fibers: implications for models of vertebrate muscle recruitment, J. Comp. Physiol., A 175 (1) (1994) 123 131. [42] L.C. Rome, P.T. Loughna, G. Goldspink, Muscle fiber activity in carp as a function of swimming speed and muscle temperature, Am. J. Physiol. 247 (16) (1984) R272 R279. [43] M.H. Laughlin, R.B. Armstrong, Muscular blood flow distribution patterns as a function of running speed in rats, Am. J. Physiol. 243 (1982) H296 H306. [44] R.L. Marsh, et al., Partitioning the energetics of walking and running: swinging the limbs is expensive, Science 303 (2004 (2 January)) 80 83. [45] P.J. Randle, et al., The glucosefatty acid cycle: its role in insulin sensitivity and the metabolic disturbances of diabetes mellitus, Lancet 1 (1963) 785 789.

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[46] M.C. Sugden, K. Bulmer, M.J. Holness, Fuel-sensing mechanisms integrating lipid and carbohydrate utilization, Biochem. Soc. Trans. 29 (2) (2001) 272 278. [47] M. Roden, How free fatty acids inhibit glucose utilization in human skeletal muscle, News Physiol. Sci. 19 (2004) 92 96. [48] N.B. Ruderman, et al., Malonyl-CoA, fuel-sensing, and insulin resistance, Am. J. Physiol. 276 (39) (1999) E1 E18. [49] D. Dean, et al., Exercise diminishes the activity of acetyl-CoA carboxylase in human muscle, Diabetes 49 (2000) 1295 1300. [50] A.E. Jeukendrup, Regulation of fat metabolism in skeletal muscle, Ann. N.Y. Acad. Sci. 967 (2002) 217 235. [51] J.M. Stewart, Fatty acid binding proteins as metabolic regulators, in: A.K. Duttaroy, F. Spener (Eds.), Cellular Proteins and Their Fatty Acids in Health and Disease, Wiley-VCHm, Weinheim, 2003, pp. 383 395.