Effects of short-term submaximal training in humans on muscle metabolism in exercise

C. T. Putman1, N. L. Jones1, E. Hultman2, M. G. Hollidge-Horvat1, A. Bonen3, D. R. McConachie1, and G. J. F. Heigenhauser1

1 Department of Medicine, McMaster University Medical Centre, Hamilton, Ontario L8N 3Z5; 3 Department of Kinesiology, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada; and 2 Department of Clinical Chemistry, Huddinge University Hospital, Karolinska Institute, S-141 86 Huddinge, Sweden

    ABSTRACT
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Abstract
Introduction
Methods
Results
Discussion
References

Muscle metabolism, including the role of pyruvate dehydrogenase (PDH) in muscle lactate (Lac-) production, was examined during incremental exercise before and after 7 days of submaximal training on a cycle ergometer [2 h daily at 60% peak O2 uptake (VO2 max)]. Subjects were studied at rest and during continuous steady-state cycling at three stages (15 min each): 30, 65, and 75% of the pretraining VO2 max. Blood was sampled from brachial artery and femoral vein, and leg blood flow was measured by thermodilution. Biopsies of the vastus lateralis were obtained at rest and during steady-state exercise at the end of each stage. VO2 max, leg O2 uptake, and the maximum activities of citrate synthase and PDH were not altered by training; muscle glycogen concentration was higher. During rest and cycling at 30% VO2 max, muscle Lac- concentration ([Lac-]) and leg efflux were similar. At 65% VO2 max, muscle [Lac-] was lower (11.9 ± 3.2 vs. 20.0 ± 5.8 mmol/kg dry wt) and Lac- efflux was less [-0.22 ± 0.24 (one leg) vs. 1.42 ± 0.33 mmol/min] after training. Similarly, at 75% VO2 max, lower muscle [Lac-] (17.2 ± 4.4 vs. 45.2 ± 6.6 mmol/kg dry wt) accompanied less release (0.41 ± 0.53 vs. 1.32 ± 0.65 mmol/min) after training. PDH in its active form (PDHa) was not different between conditions. Calculated pyruvate production at 75% VO2 max fell by 33%, pyruvate reduction to lactate fell by 59%, and pyruvate oxidation fell by 24% compared with before training. Muscle contents of coenzyme A and phosphocreatine were higher during exercise after training. Lower muscle lactate production after training resulted from improved matching of glycolytic and PDHa fluxes, independently of changes in muscle O2 consumption, and was associated with greater phosphorylation potential.

lactate; oxygen uptake; pyruvate dehydrogenase; glucose transporters; glycogen; leg blood flow; free fatty acids; phosphorylation potential

    INTRODUCTION
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Abstract
Introduction
Methods
Results
Discussion
References

LONG-TERM ENDURANCE (submaximal) training carried out over periods of at least several weeks leads to a reduction in muscle glycogen utilization and lactate production, with greater use of fats as fuel during submaximal exercise (17). These changes have been ascribed to improvements in oxygen delivery through increases in muscle capillarization, greater metabolic oxidative capacity related to increases in mitochondrial content, and increased activity of enzymes taking part in the citric acid cycle and fat oxidation. Recently, short-term (6-10 days) endurance training has also been shown to shift metabolism from carbohydrates to fats, with less lactate production, before any increases in mitochondrial oxidative capacity have occurred (15, 16, 24), calling into question the importance of increasing oxygen delivery and oxidative capacity in explaining the effects of training on the choice of fuels for exercise. Muscle biopsy and whole body substrate turnover studies have shown that muscle glycogen and glucose utilization are decreased and less lactate is produced (8, 15, 16, 21, 23, 24). Intramuscular triglycerides are oxidized to a greater extent, without increases in the oxidation of plasma-derived free fatty acids (20, 21, 23, 24).

A reduction in blood lactate accumulation is uniformly found after endurance training (17), and because traditionally lactate formation has been considered to be secondary to exercise-induced tissue hypoxia, improvements in oxygen delivery and utilization have been considered to be the causal mechanisms. An alternative view of lactate formation (29) is that it results from an imbalance between the rates of pyruvate production by glycolysis and of pyruvate oxidation by the pyruvate dehydrogenase enzyme complex (PDHc) (26) and enzymes of oxidative phosphorylation (29). Because the equilibrium constant of the lactate dehydrogenase system markedly favors lactate over pyruvate, small changes in pyruvate concentration are associated with large increases in lactate formation.

Recently, it has been demonstrated (8) that the reduction in muscle lactate production after short-term training is associated with a reduction in glycogenolytic rate, related to posttransformational downregulation of glycogen phosphorylase activity, modulated by decreases in both PO2-4 and AMP. In all these previous studies, reductions in lactate accumulation were larger than the accompanying reductions in glycogenolysis. Recent lactate tracer studies have shown that after short-term training (22), there is in addition to a lower muscle lactate production a greater rate of lactate clearance from the blood.

Because PDHc controls the rate of acetyl-CoA formation from pyruvate, the effects of training on lactate metabolism may be explained by an increase in the active form of PDHc (PDHa). Greater conversion of pyruvate into acetyl-CoA may allow greater oxidative phosphorylation and lower pyruvate concentration, accompanied by lower lactate formation. Because previous studies of short-term training have shown no increase in mitochondrial enzymes, any increases in lactate clearance and pyruvate oxidation are likely to result from altered PDHa regulation at submaximal workloads, rather than increases in the total PDHc activity (PDHt). However, the role of increased transformation of PDHc to PDHa in attenuating net muscle lactate production after short-term training has not been established. The purpose of the present study was to examine the regulation of PDHa during exercise before and after this type of training.

    METHODS
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Abstract
Introduction
Methods
Results
Discussion
References

Subjects

Seven male subjects [age 23.4 ± 1.5 (SE) yr, height 181 ± 2 cm, weight 81.4 ± 4.1 kg] participated in this study. Approval was obtained from the ethics committees of McMaster University and the McMaster University Medical Centre. Written informed consent was obtained from all subjects after an explanation of the attendant risks associated with the study protocol.

Training Protocol

Before training, subjects completed a progressive exercise test on a cycle ergometer to determine peak oxygen uptake (VO2 max) and maximum work capacity. Measurements of O2 uptake and CO2 output were made using either a metabolic cart (Quinton Q-plex 2; Quinton Instrument, Seattle, WA) or by mass spectrometry. On a separate day, the subjects were studied at rest and during three levels of continuous steady-state exercise (30, 65, and 75% of VO2 max), each maintained for 15 min. Each subject then completed either 7 or 8 consecutive days of training on a cycle ergometer at 60% of their pretraining VO2 max for 2 h daily. On the day after completion of the training period, VO2 max was again determined, and on the next day experiments were repeated at rest and at the same absolute workloads as before training.

Experimental Protocol

On the morning of each experiment, subjects ate a standard light meal, consisting primarily of carbohydrate, and reported to the laboratory 1-2 h later. The femoral vein was catheterized percutaneously by means of the Seldinger technique, after administration of 3-4 ml of xylocaine, as described by Bernéus et al. (4). The brachial artery was catheterized percutaneously (4) with a radiopaque Teflon catheter after local anesthesia with 0.5 ml of xylocaine. Catheters were maintained patent with sterile nonheparinized saline solution. Leg blood flow was determined using the thermodilution method, as described by Andersen and Saltin (1): 10 ml of nonheparinized saline were injected, and leg blood flow was determined from the change in temperature as a function of time by use of a portable CO monitor (Spacelab, Redmond, WA). At least three measures were recorded at each time point and averaged.

Before and after training, four biopsy sites were prepared superficial to the vastus lateralis after local anesthesia. Subjects remained sitting and inactive for 20 min (preexercise), followed by cycling for 15 min each at 30, 65, and 75% of their pretraining VO2 max. Respiratory measurements were made during the last 5 min of the preexercise period and between 8 and 12 min of each exercise stage. Blood samples from the femoral vein and the brachial artery were simultaneously drawn at rest and at 9 and 13 min of each exercise stage. Leg blood flow was measured immediately after blood sampling at both 9 and 13 min. Biopsies of the vastus lateralis were obtained at rest just before the start of exercise and at the end of each exercise stage, and these were immediately frozen in liquid nitrogen and stored in liquid nitrogen until analyzed.

Blood Sampling and Analysis

Arterial and venous blood samples were drawn in heparinized plastic syringes and placed on ice. One portion of each sample was deproteinized in 6% perchloric acid (PCA) and stored at -20°C until analysis for glucose, lactate, and glycerol according to the methods of Bergmeyer (3) adapted for fluorometry. The second portion of blood was immediately centrifuged at 15,900 g for 2 min; the plasma supernatant was frozen and later analyzed for free fatty acids (Wako NEFA C test kit, Wako Chemical, Montréal, QC, Canada). A third portion of blood was analyzed for O2 content (AVL 995 Automatic Blood Gas Analyzer, Intermedico, Markham, ON, Canada). Hematocrit (Hct) was determined using heparinized microcapillary tubes centrifuged for 5 min at 15,000 g.

Muscle Analysis

PDHa and PDHt were analyzed as previously described (10, 26). To compensate for contamination of muscle homogenates with blood or connective tissue, PDHa and PDHt measures were corrected to the highest total creatine in a series of biopsies obtained from each subject on a given day. Muscle glycogen was determined fluorometrically using the enzymatic end-point method described by Bergmeyer (3). Neutralized PCA extracts of freeze-dried, dissected, and powdered muscle samples were analyzed for acetyl-CoA, free CoASH, total CoA, acetylcarnitine, free carnitine, and total carnitine according to the method of Cederblad et al. (7). ATP, ADP, glucose, glucose 6-phosphate (G-6-P), fructose 6-phosphate (F-6-P), glycerol 3-phosphate (G-3-P), pyruvate, lactate, and phosphocreatine (PCr) were determined by the methods of Bergmeyer adapted for fluorometry. PCA extracts were corrected to the highest total creatine concentration within a series of biopsies obtained from each subject on a given day. Muscle wet-to-dry ratios were determined by weighing the frozen muscle samples before and again after freeze-drying. Maximal citrate synthase (CS) activity was determined according to Bergmeyer. Maximal CS activity was measured on each biopsy in duplicate on two different occasions for each subject. Because no differences were found between the two sets of independent measures, between conditions, or across time within a condition, the data were pooled and presented as pre- and posttraining. Muscle GLUT-1 and GLUT-4 transporter contents were determined by Western blot analysis (25).

Leg Uptake and Release of Metabolites and O2

Uptake and release of metabolites (glucose, free fatty acids, glycerol, and lactate) and O2 were calculated from their whole blood contents in arterial and venous blood and leg blood flow. Because there were no differences in Hct over time within a condition or between matched arterial and venous samples, blood samples were not corrected for fluid shifts. Also, because subjects were in steady state, with no significant differences in blood flows or metabolite concentrations between 9 and 13 min of each exercise stage, the two values at these time points were averaged to obtain one value for each.

Calculation of Pyruvate Production and Oxidation and Lactate Production

Pyruvate production was calculated from the sum of the rates of glycogen breakdown and glucose uptake minus the sum of the rates of accumulation of muscle glucose, G-6-P, and F-6-P by use of wet weight concentration differences and with the assumption that working muscle amounted to 4.3 kg/leg. Because net lactate release across the leg was measured during the steady state of each workload, with most lactate being produced earlier in exercise, net lactate release over the complete exercise period was instead calculated from changes in arterial blood lactate concentration by assuming a distribution volume of 0.6 × body weight. Pyruvate oxidation was calculated as pyruvate production minus lactate production.

Statistical Analysis and Summary of Data

Data were analyzed using two-way ANOVA with repeated measures over time, except where otherwise stated. When a significant F ratio was found, the Newman-Keuls post hoc analysis was used to compare means over time and between conditions. The following data were analyzed using a 2-tailed paired dependent-samples Student's t-test: relative and absolute VO2 max, body weight, maximal CS activity, PDHt, GLUT-1, GLUT-4, mean total CoA, mean total carnitine, pyruvate production, pyruvate oxidation, and lactate production. Because we specifically hypothesized that respiratory exchange ratio (RER) and glycogen utilization would decrease as a result of training, these data were analyzed using a 1-tailed paired dependent-samples Student's t-test. Data are summarized as means ± SE. Differences were considered significant at P < 0.05.

    RESULTS
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Abstract
Introduction
Methods
Results
Discussion
References

VO2 max and Respiratory Gas Exchange Responses to Training and Cycling Exercise

No changes were observed in maximum aerobic capacity as a result of submaximal training, as indicated by no changes in muscle CS activity and in absolute or relative VO2 max (Table 1). Whole body VO2 (Table 1) and O2 uptake across the leg (see Table 5) were not changed by training. In contrast, ventilation (VE) was lower at 75% VO2 max and CO2 uptake (VCO2) was lower at 65% VO2 max in the posttraining condition. After training, RER (Table 2) was lower during cycling at 30 and 65% VO2 max.

                              
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Table 1.   Maximal oxygen uptake, body weight, maximal enzyme activities, and skeletal muscle GLUT-1 and GLUT-4 glucose transporters before and after short-term training

                              
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Table 2.   Respiratory measures before and after short-term training

Muscle Metabolism

ATP, ADP, and PCr. Muscle ATP and ADP concentrations (Table 3) were unaltered by exercise or as a result of training. Before training, PCr concentration ([PCr]) decreased with each increase in exercise intensity from 30 to 75% VO2 max. However, after training, [PCr] did not decrease further from 65 to 75% VO2 max, being higher at this level of exercise than before training (Table 3).

                              
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Table 3.   Muscle metabolite measures and muscle wet-to-dry ratios before and after training

Glycogen. After training, intramuscular glycogen was greater at rest and throughout each stage of the exercise protocol (Table 3). Glycogen utilization during cycling at 30 and 65% of VO2 max was not different between the pretraining and posttraining conditions. However, at 75% VO2 max, pretraining glycogen utilization (113.2 ± 14.6 mmol/kg dry wt) was 42.5 mmol/kg dry wt greater (P < 0.02) than in the posttraining condition (70.7 ± 13.9 mmol/kg dry wt).

Glucose, G-6-P, F-6-P, and G-3-P. Intramuscular accumulation of glucose was similar between conditions at each stage, except at 75% VO2 max, where it was lower posttraining (Table 3). Muscle G-6-P and G-3-P contents were also lower after training (Table 3). F-6-P (Table 3) was not different between the pre- and posttraining conditions.

Lactate and pyruvate. Muscle lactate concentration ([lactate]) (Fig. 1) was significantly lower after training (main effect P < 0.03). Before training, muscle [lactate] increased progressively from 4.3 ± 0.5 mmol/kg dry wt at rest to 9.0 ± 3.8 at 30%, 20.0 ± 5.8 at 65%, and 45.2 ± 6.6 mmol/kg dry wt at 75% VO2 max. In the posttraining condition, muscle [lactate] did not increase above the initial preexercise value of 4.2 ± 0.4 at 30% (4.3 ± 0.3) but then increased to 11.9 ± 3.2 and 17.2 ± 4.4 mmol/kg dry wt at 65 and 75% of VO2 max, respectively. Muscle pyruvate concentration ([pyruvate]) progressively increased to the same extent, with increasing workload in both the pre- and posttraining conditions up to 65% VO2 max (Fig. 1). At 75% VO2 max, however, posttraining muscle [pyruvate] (0.57 ± 0.07 mmol/kg dry wt) was 32% lower than the pretraining value (0.75 ± 0.06 mmol/kg dry wt) (Fig. 1).


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Fig. 1.   Muscle lactate and pyruvate concentrations before and after training. VO2 max, maximal O2 uptake. + Significantly different from pretraining at matched time points. There was a significant main effect over time for posttraining lactate data and for both pre- and posttraining pyruvate data. dw, Dry wt.

PDHa. PDHt was similar before and after training (Table 1). Before training, the preexercise PDHa level (Fig. 2) was 0.87 ± 0.11 mmol · min-1 · kg wet wt-1 and increased to 1.81 ± 0.29, 3.58 ± 0.76, and 3.89 ± 0.87 mmol · min-1 · kg wet wt-1 after cycling at 30, 65, and 75% VO2 max, respectively. After training, PDHa was not different from the pretraining condition at matched time points, being 0.84 ± 0.10 mmol · min-1 · kg wet wt-1 before cycling exercise and 1.65 ± 0.24, 2.60 ± 0.13, and 3.78 ± 0.36 mmol · min-1 · kg wet wt-1 at 30, 65, and 75% VO2 max, respectively.


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Fig. 2.   Muscle pyruvate dehydrogenase activity (PDHa) before and after training. There was a significant main effect over time for both pre- and posttraining conditions. ww, Wet wt.

CoA, carnitine, and acetylated forms. Total muscle CoA content increased as a result of short-term training (Table 4). When averaged across all four biopsies within each condition, this amounted to a 25% increase (P < 0.03) in the mean total CoA content posttraining (pre- vs. posttraining: 75.4 ± 5.4 vs. 94.6 ± 11.2 mmol/kg dry wt). Acetyl-CoA increased to the same extent with increasing exercise intensity in both conditions (Table 4). Free CoASH behaved in a reciprocal fashion, decreasing as a function of increasing exercise intensity (Table 4). Total muscle carnitine content was not altered by cycling exercise or by training (Table 4), the averages of four biopsies taken before and after training being 20.3 ± 1.6 and 20.3 ± 2.0 mmol/kg dry wt, respectively. Acetylcarnitine followed a similar pattern as acetyl-CoA, increasing as a function of increasing exercise intensity, with the exception that acetylcarnitine accumulation was attenuated at 65 and 75% VO2 max posttraining (Table 4). Free carnitine decreased in a reciprocal manner and did not differ between conditions, except at 65% VO2 max posttraining, when it was greater.

                              
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Table 4.   Muscle CoA, carnitine, and their acetylated forms before and after training

Metabolite Exchange Across the Leg

Leg blood flow and O2 uptake. Leg blood flow progressively increased from preexercise (i.e., rest) to 65% VO2 max to the same extent in the pre- and posttraining conditions (Table 5). However, at 75% VO2 max, leg blood flow was 0.84 l/min greater after training. O2 uptake across the leg was not altered by training (Table 5).

                              
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Table 5.   Arterial concentrations of blood-borne substrates, leg blood flow, and leg O2 uptake before and after training

Blood lactate. At rest and during cycling at 30% VO2 max, arterial [lactate] (Table 5) did not differ within or between conditions, but at 65 and 75% VO2 max, there was a progressive increase in both conditions. However, after training, arterial [lactate] values were lower at 65 and 75% VO2 max (Table 5).

During rest and cycling at 30% VO2 max, there were no differences in the net release of lactate between the pre- and posttraining conditions (Fig. 3). At 65 and 75% VO2 max, net lactate release was less at posttraining (Fig. 3).


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Fig. 3.   Glucose uptake and lactate release across the leg before and after training. + Significantly different from pretraining at matched time points. There was a significant main effect over time for both pre- and posttraining glucose data and posttraining lactate data.

Blood glucose and muscle glucose transporters. Arterial glucose concentration ([glucose]) was similar at all power outputs in both pre- and posttraining conditions (Table 5). Leg glucose uptake (Fig. 3) did not differ between conditions. Training resulted in a 43% increase in GLUT-4 glucose transporter content but no change in GLUT-1 content (Table 1).

Blood free fatty acids and glycerol. Arterial free fatty acid (FFA) concentrations were not altered by training or as a result of the exercise protocol (Table 5). No changes were observed between conditions in the net uptake or release of FFA before and during cycling exercise (Fig. 4).


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Fig. 4.   Free fatty acid uptake and glycerol release across the leg before and after training.

Arterial glycerol concentration increased with increasing exercise intensity, but there were no differences between the pre- and posttraining conditions (Table 5). Similarly, there were no significant differences in glycerol release between conditions (Fig. 4).

Pyruvate production, oxidation, and conversion to lactate. During cycling exercise at 75% VO2 max, pyruvate production was reduced by 33% posttraining, whereas pyruvate oxidation decreased by 24%; lactate production was reduced by 59% (Table 6).

                              
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Table 6.   Pyruvate metabolism at 75% VO2 max before and after training

    DISCUSSION
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Abstract
Introduction
Methods
Results
Discussion
References

The present study examined the effects of short-term training on some biochemical responses to 45 min of continuous exercise, consisting of 15 min each at 30, 65, and 75% VO2 max. Although there were no significant differences in glycogen utilization and lactate production at 30% VO2 max, lactate accumulation in muscle and blood was lower at 65 and 75% VO2 max after training. The most marked changes associated with training were observed at the highest load; there was a 33% decrease in pyruvate production, a 59% decrease in pyruvate conversion to lactate, and a 24% decrease in pyruvate oxidation (Table 6). The decrease in pyruvate production at this load was explained by a 30% decrease in glycogen degradation and a small, nonsignificant reduction in glucose utilization. Pulmonary and leg O2 uptake and the activity of mitochondrial CS were unchanged. The total activity of the PDHc and the proportion in its active form (PDHa) were unchanged. An increase in the total muscle content of CoA was observed, and [PCr] was also higher after training.

Previous studies of short- and long-term training have suggested that the decrease in muscle and blood lactate concentrations is the result of both a decrease in lactate production from glycolysis and an increase in its metabolic clearance from the circulation. Isotope tracer studies in rats (11, 12) and humans (19, 22) have demonstrated that reductions in blood lactate with training are associated with increases in clearance of lactate from the plasma pool rather than reductions in its appearance into plasma from muscle. In the present study, after training, lower blood and muscle [lactate] values at 75% VO2 max during steady state were associated with a lower rate of glycolysis, and efflux of lactate from muscle (appearance) was associated with a smaller increase in pyruvate oxidation (clearance). The lower activation of glycogenolysis and glycolysis posttraining may be related to the higher [PCr], itself an indication of higher phosphorylation potential of the high-energy phosphate pool (ATP, ADP, AMP, and PCr), concomitant with a lower PO2-4 concentration (8, 15, 22, 24). PO2-4 is known to act as a substrate for phosphorylase a, having a relatively high Michaelis-Menten constant that is decreased by increases in AMP concentration (27). The phosphorylation potential will thus have a direct effect on the rate of glycogen degradation, as well as a direct influence on the activity of phosphofructokinase via the ATP/ADP ratio.

An increase in phosphorylation potential has been observed, both after short-term training (8, 15, 22, 24) and when the availability of FFA for muscle metabolism is increased by infusion of fat emulsion (13) and caffeine ingestion (28). In all these situations, increased [PCr] and decreased calculated concentrations of free ADP and AMP have been accompanied by glycogen sparing during exercise. Similar findings were reported by From et al. (14) in studies of heart muscle. Thus, when FFA availability is increased, oxidative phosphorylation is enhanced, glycogen utilization and lactate formation decrease, and there is a lower PCr degradation.

A reduction in glycogenolytic rate during exercise after training was associated with reductions in pyruvate oxidation. As this reduction was unaccompanied by changes in PDHt (Table 1) or PDHa (Fig 2), we infer that it is due to a reduction in pyruvate availability secondary to a lower rate of glycogenolysis. The decrease in pyruvate oxidation may be compensated for by an increase in FFA oxidation. This argument is supported by calculations of the amount of FFA (palmitate) that would be needed to account for this difference in oxidative ATP production, showing that only 4.5 mmol of palmitate would be required for each leg during 15 min of exercise at 75% VO2 max posttraining. Small increases in fat oxidation may not have a significant effect on measured RER (seen at 30 and 65% VO2 max but not at 75% VO2 max), because this measurement is too insensitive to reveal such small increases in fat oxidation. Increases in fat oxidation during exercise posttraining have been observed in many studies (17, 20), including those employing short periods of training (8, 9, 21, 23, 24). The 25% increase in total CoA observed in the present study may have an impact on fatty acid utilization, because both the acylation and the transport of fatty acids are dependent on CoA availability.

The results of isotope studies carried out pre- and posttraining (19, 22) were interpreted to indicate an increase in pyruvate oxidation posttraining. Because PDHa catalyzes the flux-generating step of pyruvate oxidation by the TCA cycle (2), and in view of the isotope studies, we hypothesized that lower net lactate production after training resulted from an increase in PDHa. Although we did not find changes in PDHa to account for reductions in lactate accumulation posttraining, our measurements were made after 15 min of cycling at a given power; thus it is still possible that PDHa or the allosteric regulators of PDHc transformation were altered during the transition to a steady state, from rest to exercise or from one load to a higher one. Recently, Timmons and co-workers (30, 31) employed the isolated dog gracilis model to study the effects of increases in the rate of PDH activation by dichloroacetate at the onset of muscle contraction. They found that PDH activation, with concomitant increases in acetylcarnitine concentration before muscle stimulation, resulted in less lactate production during ischemic contractions. However, in the present study after training, acetylcarnitine accumulation was less just before the onset of the highest exercise intensity (75% VO2 max), in which the reduction in lactate production was greatest. Thus the association between increases in fat utilization and reductions in lactate production after short-term training observed in the present study may be mediated through different mechanisms from those in the animal model used by Timmons et al. (31).

Isotopic studies of the effects of training suggested that training was associated with an increase in lactate clearance from the plasma pool, with no change in the appearance rate of lactate into the pool. This finding is in contrast to the results of the present study, which utilized measurements of intramuscular concentration of metabolites with arteriovenous lactate concentrations and flow. These measurements suggested that there was both a reduction in muscle lactate production and a lower efflux of lactate from muscle. A narrowing of the concentration gradient for lactate between arterial blood and muscle is presumably accounted for by an increase in the content of the sarcolemmal monocarboxylate transporter (MCT1) found in the present subjects and reported elsewhere (5). This increase in MCT1 would increase bidirectional exchange of lactate between the plasma and intracellular compartments and also account for the increases in lactate clearance from the plasma pool that were observed in isotope studies.

In the present study, leg blood flow during exercise was increased by training (Table 5). This finding is consistent with increased capillary density associated with type IIa fibers recently reported (16), which provides a morphological basis for the physiological change observed in our subjects (Table 5). Thus it is possible that the uptake of exogenous blood-borne substrates and/or endogenous fat utilization may be enhanced during the non-steady-state period at the onset of cycling exercise after training.

Previous studies using short-term training to examine the early adaptations of muscle glucose metabolism during submaximal aerobic cycling have reported greater GLUT-4 content (25) and maximal hexokinase activity (9, 24). Although such changes should serve to increase plasma glucose uptake and utilization, tracer studies (9, 21, 23) have paradoxically reported a reduction in whole body plasma glucose turnover during cycling exercise after training. Lower rates of glucose appearance from hepatic gluconeogenesis and disappearance from the plasma pool were thought to result from lower hormonally induced hepatic glucose production and lower glucose utilization by trained muscle (9, 21, 23), respectively. However, the use of labeled glucose tracers to determine glucose turnover may underestimate glucose utilization, because labeled glucose can be converted to metabolites that are not oxidized (18). Furthermore, short-term training may increase this conversion, creating the impression of lower glucose utilization posttraining. In the present study, there were no differences in the preexercise rates of glucose uptake (Fig. 3), arterial glucose concentrations (Table 5), or muscle glucose content (Table 3), but muscle glycogen levels (Table 3) were considerably greater posttraining. Whereas there were no changes in the content of the GLUT-1 glucose transporter (Table 1), GLUT-4 content (Table 1) increased by 43% after training. Thus greater preexercise glycogen levels posttraining can be attributed to enhanced glycogen synthesis mediated by greater insulin-stimulated glucose uptake by GLUT-4 and greater maximal activities of hexokinase and glycogen synthase during the recovery periods between each training session. This suggestion is supported by a previous report demonstrating similar changes after 7 consecutive days of chronic low-frequency electrical stimulation (6).

Summary and Conclusions

This study examined mechanisms of altered muscle lactate production during cycling exercise in humans after 7-8 days of training on a cycle ergometer at 60% VO2 max. Training resulted in a significant reduction in net muscle lactate production during steady-state cycling exercise at 65 and 75% VO2 max, without change in muscle oxygen consumption during exercise or maximum muscular oxidative potential. Lower muscle lactate production after training resulted from the attenuation of glycogenolysis without a change in PDHa, leading to improved matching of glycolytic and PDHa fluxes. Changes leading to lower muscle lactate production coincided with improved maintenance of cell phosphorylation potential and greater use of fat as a fuel.

    ACKNOWLEDGEMENTS

We thank T. Bragg, R. Davidson, C. French, Dr. M. Ganagaragah, and G. Obminski for excellent technical assistance with these experiments.

    FOOTNOTES

This study was supported by the Canadian Medical Research Council (MRC), the Heart and Stroke Foundation of Ontario, the Natural Sciences and Engineering Research Council of Canada, and the Canadian Diabetes Association. C. T. Putman and M. G. Hollidge-Horvat were supported by Studentships from the MRC. G. J. F. Heigenhauser is a Career Investigator of the Heart and Stroke Foundation of Ontario.

Present address of C. T. Putman: Faculty of Biology, University of Konstanz, Box M641, D-78457 Konstanz, Germany.

Address for reprint requests: G. J. F. Heigenhauser, Dept. of Medicine, McMaster Univ. Medical Centre, 1200 Main St. West, Hamilton, ON, Canada L8N 3Z5.

Received 15 April 1997; accepted in final form 8 April 1998.

    REFERENCES
Top
Abstract
Introduction
Methods
Results
Discussion
References

1.   Andersen, P., and B. Saltin. Maximal perfusion of skeletal muscle in man. J. Physiol. (Lond.) 366: 233-249, 1985[Abstract].

2.   Behal, R. H., D. B. Buxton, J. G. Robertson, and M. S. Olson. Regulation of the pyruvate dehydrogenase multienzyme complex. Annu. Rev. Nutr. 13: 497-520, 1993[Medline].

3.   Bergmeyer, H. U. Methods of Enzymatic Analysis. New York: Academic, 1983.

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