Acid proteolytic capacity in mouse cardiac and skeletal muscles after prolonged submaximal exercise

Acid proteolytic capacity in mouse cardiac muscle and in predominantly white (distal head of m. vastus lateralis) or predominantly red (proximal red heads of m. vastus lateralis, m. v. medialis, and m. v. intermedius) skeletal muscle was estimated 5 days after 3 h, 6 h or 9 h prolonged running at a speed of 13.5 m/min. The activities of acid protease and -glucuronidase together with the rate of acid autolysis considerably increased in both skeletal muscle types, especially in red muscle, but did not increase in cardiac muscle. Acid proteolytic capacity and -glucuronidase activity increased in relation to the duration of running. Protein content and oxidative capacity (the activities of citrate synthase and malate dehydrogenase) decreased in red skeletal muscle after 6 h and 9 h running. In white muscle only protein content slightly decreased after 9 h running. No corresponding changes were observed in cardiac muscle. Histopathological changes were traced in mixed skeletal muscle (m. rectus femoris). Necrotic lesions were observed in the red superficial area of m. rectus femoris after 6 h and, in particular, after 9 h running. The results show that prolonged submaximal running also produces lethal and sublethal skeletal muscle fibre injuries, as well as exhaustive exercise or temporary ischaemia as reported earlier. It is suggested that sublethal injuries precede lethal ones and that acid proteolytic capacity increases especially in the sublethally injured muscle fibres.     

Acid proteolytic capacity in mouse cardiac and skeletal muscles after prolonged submaximal exercise

Acid proteolytic capacity in mouse cardiac muscle and in predominantly white (distal head of m. vastus lateralis) or predominantly red (proximal red heads of m. vastus lateralis, m. v. medialis, and m. v. intermedius) skeletal muscle was estimated 5 days after 3 h, 6 h or 9 h prolonged running at a speed of 13.5 m/min. The activities of acid protease and β-glucuronidase together with the rate of acid autolysis considerably increased in both skeletal muscle types, especially in red muscle, but did not increase in cardiac muscle. Acid proteolytic capacity and β-glucuronidase activity increased in relation to the duration of running. Protein content and oxidative capacity (the activities of citrate synthase and malate dehydrogenase) decreased in red skeletal muscle after 6 h and 9 h running. In white muscle only protein content slightly decreased after 9 h running. No corresponding changes were observed in cardiac muscle. Histopathological changes were traced in mixed skeletal muscle (m. rectus femoris). Necrotic lesions were observed in the red superficial area of m. rectus femoris after 6 h and, in particular, after 9 h running. The results show that prolonged submaximal running also produces lethal and sublethal skeletal muscle fibre injuries, as well as exhaustive exercise or temporary ischaemia as reported earlier. It is suggested that sublethal injuries precede lethal ones and that acid proteolytic capacity increases especially in the sublethally injured muscle fibres.     

Lysosomal changes related to exercise injuries and training-induced protection in mouse skeletal muscle

Three experiments were designed to study the lysosomal changes associated with the development and maintenance of the endurance training induced resistance against exercise injuries in mouse skeletal muscles. The activities of arylsulphatase, cathepsin C, cathepsin D, and β-glucuronidase were assayed from the red part of mouse quadriceps femoris muscle 4 days after prolonged strenuous running of 4–9 h duration. Exercise injuries were characterized by necrotic fibers and focal inflammation. Strenuous running of untrained mice induced necrotic lesions and a 4–5 fold increase in the activities of lysosomal enzymes. This lysosomal response was considerably reduced already by daily training bouts on the 3 days preceding the strenuous exertion. Simultaneously exercise injuries were markedly reduced. Extending the endurance training program increased the running ability of mice and further reduced the necrotic lesions and lysosomal changes induced by the strenuous exercise. The detraining of 1 week after the termination of regular endurance training considerably increased the degree of exercise induced lysosomal response. The detraining of longer durations further increased the lysosomal response and no effect of prior endurance training existed after 1 month detraining. Our observations suggest that the severity of exercise injuries is related to the strength of the exercise stimulus and the level of preceding physical activity and can be characterized by the lysosomal changes.     

Exercise myopathy: selectively enhanced proteolytic capacity in rat skeletal muscle after prolonged running

The proteolytic capacity of rat skeletal muscle was analyzed during the repair of fiber injuries after strenuous exercise. A single bout of prolonged exercise (8 hr running at a speed of 17 m X min-1) caused a slight fiber necrosis and a selective response in the proteolytic activity of rat skeletal muscle. Acid proteolytic capacity (cathepsin D and acid autolysis) was considerably increased on the 4th day after exertion and partially decreased by the 10th day. The acid hydrolytic response was more prominent in red than in white skeletal muscle. Alkaline proteolytic capacity (alkaline and myofibrillar proteases), increased in several atrophic myopathies, was not affected in exercise myopathy. The rate of neutral autolysis slightly increased after exertion. The protein content of skeletal muscle was decreased on the 4th day after exertion. We suggest that the proteolytic responses to acute injuries, as well as to chronic atrophies, are highly selective in skeletal muscle fibers.     

Exercise-induced necrotic muscle damage and enzyme release in the four days following prolonged submaximal running in rats

Male Wistar rats were made to run uphill on a treadmill 5.5° incline at 17 m min^–1 for 4 h, and killed for muscle and serum sampling 2, 4, 12, 24, 48 or 96 h after the exertion. To estimate the degree of muscle damage,-glucuronidase activity, total protein concentration, water content and morphology were examined in the red parts of quadriceps femoris (MQF) and soleus (MS) muscles, the distal white part of the rectus femoris muscle (MRF) and the superficial part of triceps brachii muscle (MTB). Simultaneous serum samples were assayed for creatine kinase (CK) activity and carbonic anhydrase III (CA III) concentration. Fibre swelling and interstitial oedema were detected in MS at 4 h and in MQF at 12 h and typical histopathological changes, including inflammation and fibre necrosis, in both muscles 12–96 h post-exertion.-Glucuronidase activity, a quantitative marker of muscle damage, was increased in MS at 4 h, in MQF at 24 h and in MRF 48 h after the running. No increase occurred in MTB. Water and protein content increased or decreased respectively, faster in MS (2 h post-exercise) than in MQF (12 h) or MRF (12 h). Water content thus contributed to muscle damage by preceding the increase in-glucuronidase activity. Serum CK activity was increased 2, 4, and 48 h after the running. Changes in serum CA III concentration were rather similar to those in CK but were not significant. The increase in serum CK was not in concert with the necrotic events in the muscle but occurred considerably earlier (2 h vs. 12–24 h post-exercise). The second peak in CK, 48 h post-exercise (during the necrotic phase), was smaller than the first one. Our results show that serum CK activity is an inaccurate estimate of exercise-induced muscle damage as regards interpretation of the degree and the time course of pathological events in the muscle.     

Increased mRNAs for procollagens and key regulating enzymes in rat skeletal muscle following downhill running

 The purpose of the study was to investigate pre-translational regulation of collagen expression after a single bout of exercise. We analysed steady-state messenger ribonucleic acid (mRNA) levels for collagen types I, III and IV, α- and β-subunits of prolyl 4-hydroxylase and lysyl oxidase (enzymes modifying procollagen chains), and enzyme activity of prolyl 4-hydroxylase from rat soleus muscle (MS) and the red parts of quadriceps femoris muscle (MQF) after 12 h and after 1, 2, 4, 7 and 14 days of downhill (–13.5°) treadmill running at a speed of 17 m·min^–1 for 130 min. Histological and biochemical assays revealed exercise-induced muscle damage in MQF but not MS. Steady-state mRNA levels for the α- and β-subunits of prolyl 4-hydroxylase in MQF, lysyl oxidase in MS and MQF were increased 12 h after running, whereas prolyl 4-hydroxylase activity did not increase until 2 days after exercise. The mRNA levels for the fibrillar collagens (I and III) and basement membrane type IV collagen significantly increased 1 day and 12 h after exertion, respectively. Peak mRNA levels were observed 2–4 days after running, the increases being more pronounced in MQF than in MS. No significant changes were observed in types I or III collagen at the protein level. Strenuous downhill running thus causes an increase in gene expression for collagen types I and III and their post-translational modifying enzymes in skeletal muscle in a co-ordinated manner. These changes, together with the increased gene expression of type IV collagen, may represent the regenerative response of muscle extracellular matrix to exercise-induced injury and an adaptive response to running exertion. Received: 20 July 1998 / Received after revision: 30 November 1998 / Accepted: 26 January 1999     

Endurance training decreases the alkaline proteolytic activity in mouse skeletal muscles

Summary Alkaline and myofibrillar protease activities of rectus femoris, soleus, and tibialis anterior muscles and the pooled sample of gastrocnemius and plantaris muscles were analyzed in male NMRI-mice during a running-training program of 3, 10, or 20 daily 1-h sessions. The activity of citrate synthase increased during the endurance training, reflecting the increased oxidative capacity of skeletal muscles. The activities of alkaline and myofibrillar proteases continually decreased in the course of the training program in all muscles studied. Instead, the activity of-glucuronidase (a marker of lysosomal hydrolases) increased in all muscles. The highest activities were observed at the beginning of the training program. Present results, together with our earlier observations, show that the type of training, running as opposed to swimming, modulates the training responses in alkaline protease activities. Further, diverse adaptations in the activities of alkaline proteases and a lysosomal hydrolase suggest differences in the function of different proteolytic systems.This study was supported by the Academy of Finland and the Research Council for Physical Education and Sport (Ministry of Education, Finland)     

Food deprivation decreases the exertion-induced acid hydrolase response in mouse skeletal muscle

Summary Strenuous prolonged running causes muscle fibre necrosis in skeletal muscles. The muscle injury is associated with inflammation and a strong increase in the total activities of certain acid hydrolases a few days after exertion. The activity changes of acid hydrolases quantitatively well reflect the severity of histopathological changes during the myopathy (for review see Salminen, Acta Physiol Scand [Suppl 539] 1985). In this study male NMRI-mice were exposed to a protocol of fasting and refeeding together with or without a 6 h run on a treadmill at 13.5 m · min^–1 The animals were killed 4 days after the exercise and samples from the red part of quadriceps femoris were analyzed for arylsulfatase (ASase) and -glucuronidase (GUase) activities. Starvation protocols did not affect ASase or GUase. Running caused a 3.2-fold increase in ASase and a 5.1-fold increase in GUase. If mice were exercised in the fasted condition a normal exercise response occurred in both activities, but when mice were exercised 2 days after the finish of fasting the exercise response was greatly diminished. Thus food deprivation followed by 2 days refeeding induces a protection against exercise myopathy in mice. The protection greatly resembles that induced by regular endurance training preceding strenuous prolonged exertion.     

Acid hydrolase activities in mouse cardiac and skeletal muscle following exhaustive exercise

Summary Acid hydrolase activities in skeletal and cardiac muscle were studied 5,10, and 20 days after exhaustive intermittent running by untrained and endurance-trained mice. Exhaustion increased the activities of cathepsin D, -glucuronidase and ribonuclease, but not that of p-nitrophenylphos-phatase in skeletal muscle of untrained mice. Activities were highest on the fifth day after exhaustion and decreased during the following two weeks. More intensive loading produced no changes in acid hydrolytic capacity in skeletal muscle of endurance-trained mice. Acid hydrolase activities in cardiac muscle of both untrained and trained mice were unaffected by exhaustive running. It is suggested that exhaustive running causes both lethal and sublethal hypoxic fiber injuries in the skeletal muscle of untrained mice but not in that of endurance-trained mice or in the cardiac muscle of animals of either group. These injuries manifest themselves as fiber necrosis (lethal) and as increased acid hydrolytic capacity in surviving fibers (sublethal).This study was financially supported by the Academy of Finland and the Finnish Research Council for Physical Education and Sport (Ministry of Education)     

Latencies and phosphomannosyl-enzyme receptors of lysosomal enzymes during the appearance and repair of exercise injuries in mouse skeletal muscles

Latencies and phosphomannosyl-enzyme receptors of lysosomal enzymes were studied in the skeletal muscles of NMRI mice during the appearance (0–1 days) and the repair (3–9 days) of muscle fiber injuries after a single bout of prolonged running (9 hr, 13.5 m/min). The unsedimentable, releasable, and bound activities of arylsulfatase, β-N-acetylglucosa-minidase, β-d-glucuronidase, cathepsin C, and ribonuclease as well as the content and occupancy of phosphomannosyl-enzyme receptors of lysosomal enzymes were assayed. The distribution of enzyme activities in different fractions as well as the changes after exertion greatly varied between different lysosomal enzymes. In general, the total activities and also the distribution of enzyme activities in different fractions were unaffected 1 hr after exertion, but on the day after exertion small increases were observed in the free and releasable activities. The highest enzyme activities both in the homogenate and in different fractions were recorded 3 days after exertion, after which the activities slowly decreased. The increases of enzyme activities were higher in the free and releasable fractions than in the homogenate but the changes in the proportional distributions of lysosomal enzyme activities between different fractions were minor. The present study also showed the presence of phosphomannosyl-enzyme receptors of lysosomal enzymes in the membranes of skeletal muscles. The total content of phosphomannosyl-enzyme receptors was unchanged 0–3 days after exertion but a small increase occurred 5–8 days after exertion. Instead, the occupancy of these lysosomal receptors with endogenous enzymes was significantly increased 1–5 days after exertion and decreased later to the control level.     

Effects of the protease inhibitor leupeptin on proteolytic activities and regeneration of mouse skeletal muscles after exercise injuries.

Leupeptin, a nontoxic thiol protease inhibitor, has been proposed to have therapeutic use in hereditary muscular dystrophies. The purpose of this study was to characterize the in vivo changes in proteolytic activity of skeletal muscles induced by the repeated administration of leupeptin. Further, whether the modulation of proteolytic capacity by leupeptin affects the repair process of muscle injuries caused by heavy exercise was studied. Leupeptin was administered in mice intraperitoneally at a dose level of 15.5 mg/kg twice a day for 9 days. Leupeptin, known to be an inhibitor of cathepsin B both in vitro and after a single injection in vivo, paradoxically induced an increase of cathepsin B activity in mouse skeletal muscles after repeated administration. In addition, leupeptin administration for 9 days increased the activities of cathepsins C and D, as well as the rate of acid autolysis. The activity of beta-glucuronidase also increased, while those of arylsulfatase, ribonuclease, and alkaline protease were unaffected. No histopathologic changes were observed. At the low dosage used, leupeptin had no effect on the repair process of skeletal muscle after exercise injuries, although several proteolytic processes occur during the regeneration. It is suggested that the increase of acid protease activities in skeletal muscles is an adaptive response to the administration of the proteolytic inhibitor leupeptin and that leupeptin can be administered without prevention or delay of regenerative processes after the onset of myopathic changes.     

Effect of endurance training on the capacity of red and white skeletal muscle of mouse to oxidize carboxyl-14C-labelled palmitate.

Three groups of mice were trained for 1, 4 and 5 months according to different running programs on a motor driven treadmill and the fatty acid oxidation capacity (FAO) and the activities of some enzymes of energy metabolism (cytochrome c oxidase, malate dehydrogenase, triosephosphate dehydrogenase, and lactate dehydrogenase) were determined from m. quadriceps femoris (MQF). Endurance training increased the FAO [5-month training 4 days/week, 30 min/day 22% (p less than 0.05); 1-month training, 7 days/week, 150 min/day 37% (p less than 0.001); 4-month training, 5 days/week, 60 min/day 24% (p less than 0.05)]. The activities of cytochrome c oxidase and malate dehydrogenase increased approx. 30% (p less than 0.001) whereas triosephosphate dehydrogenase and lactate dehydrogenase activities were not prominently influenced by training. The predominantly red part of MQF of untrained animals oxidized palmitate four times faster than the predominantly white part. The activities of cytochrome c oxidase and malate dehydrogenase were two times higher showing pronounced FAO in the red part. Endurance training increased the FAO and activities of oxidative enzymes in the red and white parts and in the whole muscle relatively equally resulting in similar differences between the muscle types after training. The absolute increase in the FAO of the red muscle was, however, manyfold when compared in chemical units to the white muscle.     

Enzyme activities of energy metabolism in skeletal muscle, and in the uterus, during late pregnancy in the mouse

Three hindleg muscles (white and red quadriceps, soleus), and uterine tissue, from late pregnant NMRI mice were analyzed for selected enzyme activities of energy metabolism. In addition, skeletal muscle fibre type distribution and fibre diameters were estimated in some animals. No differences between pregnant and non-pregnant animals were found for any enzyme in any of the tissues. Neither were there any differences in muscle morphology. It was concluded, that the increased work load, and altered plasma substrate profile, during late pregnancy do not induce metabolic or morphological adaptations in skeletal muscle. Furthermore, the smooth muscle of the murine myometrium does not increase its metabolic capacity in preparation for parturition, suggesting that the base-line metabolic capacity is sufficient for the work involved. The morphological and metabolic characteristics of the three skeletal muscles were similar to those in the rat. Compared with these muscles, the uterus had a low metabolic capacity, and appeared to rely predominantly on fatty acid oxidation.     

Collagen metabolism of mouse skeletal muscle during the repair of exercise injuries

The activities of prolyl 4-hydroxylase and -glucuronidase, the concentration of hydroxyproline as well as reticulin and collagen type III, IV and V stainings were followed in skeletal muscle during a 20-day period after a 9-h treadmill running in untrained and trained male mice, aged 4–6 months. The prolonged 9-h running of untrained mice temporarily increased prolyl 4-hydroxylase activity 2, 5 and 10 days after exercise, more prominently in the red than in the white part of quadriceps femoris-muscle, and in analogical manner as -glucuronidase activity in tibialis anterior-muscle. Twenty days after exercise these enzymatic activities were back to the control level. The hydroxyproline content of red muscle was increased for 10 and that of white muscle for 20 days after the exertion. Training for 45 days did not affect hydroxyproline content and prolyl 4-hydroxylase activity was at the control level after the training. A 9-h exercise increased prolyl 4-hydroxylase activity much less in trained muscle than in the untrained muscle and did not affect muscle collagen content.Histological observations showed fiber necrosis 2 days and signs of fiber regeneration 5 days after the exertion in untrained mice. Twenty days afterwards the regeneration was nearly completed. Reticulin staining was increased in injured muscle areas 10–20 days after the exertion. In immunohistochemical staining, antibodies to all studied collagen types (type III, IV and V) showed increased staining 5–20 days after the exertion in the areas of muscle injuries and regeneration. It is concluded that collagen metabolism is stimulated during the regeneration of muscle fibers and that preceeding endurance training is able to alleviate exrcise induced injuries.     

Disparity in regional and systemic circulatory capacities: do they affect the regulation of the circulation?

In this review we integrate ideas about regional and systemic circulatory capacities and the balance between skeletal muscle blood flow and cardiac output during heavy exercise in humans. In the first part of the review we discuss issues related to the pumping capacity of the heart and the vasodilator capacity of skeletal muscle. The issue is that skeletal muscle has a vast capacity to vasodilate during exercise [∼300 mL (100 g)⁻¹ min⁻¹], but the pumping capacity of the human heart is limited to 20–25 L min⁻¹ in untrained subjects and ∼35 L min⁻¹ in elite endurance athletes. This means that when more than 7–10 kg of muscle is active during heavy exercise, perfusion of the contracting muscles must be limited or mean arterial pressure will fall. In the second part of the review we emphasize that there is an interplay between sympathetic vasoconstriction and metabolic vasodilation that limits blood flow to contracting muscles to maintain mean arterial pressure. Vasoconstriction in larger vessels continues while constriction in smaller vessels is blunted permitting total muscle blood flow to be limited but distributed more optimally. This interplay between sympathetic constriction and metabolic dilation during heavy whole-body exercise is likely responsible for the very high levels of oxygen extraction seen in contracting skeletal muscle. It also explains why infusing vasodilators in the contracting muscles does not increase oxygen uptake in the muscle. Finally, when ∼80% of cardiac output is directed towards contracting skeletal muscle modest vasoconstriction in the active muscles can evoke marked changes in arterial pressure.     

Effects of endurance training on alkaline protease activities in rat skeletal muscles

This study aimed at comparing the effects of running and swimming training protocols and the termination of training on the activities of two proteases with alkaline pH-optima (alkaline protease and myofibrillar protease) in the tibialis anterior, soleus, and gastrocnemius muscles of male rats. The training on treadmill decreased the activities of alkaline and myofibrillar proteases by approx. 10-20% in the muscles studied. The activities of both proteases were unchanged in swimming-trained rats. Two weeks after the termination of running training the activity of alkaline protease was increased in gastrocnemius muscle but not in the other muscles. Swimming training increased the activity of citrate synthase in all muscles studied but training by running only in the soleus muscle. The running protocol increased the activity of beta-glucuronidase in the tibialis anterior muscle and decreased the activity in the gastrocnemius muscle. The swimming program did not affect beta-glucuronidase activities. These results show diverse effects of running and swimming training on alkaline proteolytic activities as well as on mitochondrial and lysosomal marker enzymes.     

Skeletal muscle and heart antioxidant defences in response to sprint training

Although endurance training enhances the antioxidant defence of different tissues, information on the effect of sprint training is scanty. We examined the effect of sprint training on rat skeletal muscle and heart antioxidant defences. Male Wistar rats, 16–17 weeks old, were sprint trained on a treadmill for 6 weeks. Total glutathione levels and activities of glutathione peroxidase, glutathione reductase, glutathione S-transferase and superoxide dismutase in heart and various skeletal muscles were compared in trained and control sedentary animals. Lactate dehydrogenase and citrate synthase enzyme activities were measured in muscle to test the effects of training on glycolytic and oxidative metabolism. Sprint training significantly increased lactate dehydrogenase activity in predominantly fast glycolytic muscles and enhanced total glutathione contents of the superficial white quadriceps femoris, mixed gastrocnemius and fast-glycolytic extensor digitorum longus muscles. Oxidative metabolic capacity increased in plantaris muscle only. Compared with the control group, glutathione peroxidase activities in gastrocnemius, extensor digitorum longus muscles and heart also increased in sprint trained rats. Glutathione reductase activities increased significantly in the extensor digitorum longus muscle and heart. Glutathione S-transferase activity was also higher in the sprint trained extensor digitorum longus muscle. Sprint training did not influence glutathione levels or glutathione-related enzymes in the soleus muscle. Superoxide dismutase activity remained unchanged in skeletal muscle and heart. Sprint training selectively enhanced tissue antioxidant defences by increasing skeletal muscle glutathione content and upregulating glutathione redox cycle enzyme activities in fast and mixed fibre leg muscles and heart.     

Acute exhaustive exercise changes the metabolic profiles in slow and fast muscles of rat

The effects of acute endurance running on the metabolic profiles of rat skeletal muscle were studied. Male Wistar strain rats were continuously run on a treadmill for 1 h (speed, 35 m/min; grade, 0 degrees). Soleus (SOL) and extensor digitorum longus (EDL) were removed after 30-min running, and a 0, 1, 6, 24, 48, and 72 h post-exercise, and enzymes activity (CK, LDH, PFK, PK, SDH, and MDH) and substrates contents (glycogen and pyruvate) were measured biochemically. The time course of the enzyme activities showed two distinct patterns: CK, LDH, SDH, and MDH showed two peaks, at 0 and 24 h post-exercise, while PFK and PK showed one peak at 0 h post-exercise. The activities of glycolytic enzymes and CK in EDL and oxidative enzymes in SOL showed marked changes after exercise. The glycogen level was lowest at 0 h post-exercise in both muscles and recovered to resting level by 24 h post-exercise. Pyruvate increased with running and showed the highest value at 1 h post-exercise. Increased oxidative capacity of skeletal muscle in response to the acute endurance exercise dropped gradually to the resting level by 48 h post-exercise. An endurance exercise may induce a flexible adaptation on the oxidative capacity within skeletal muscle. We conclude that the respective time course of the enzyme activities must be considered when discussing metabolic changes that occur with acute endurance exercise.     

Differences in leg muscle activity during running and cycling in humans

Delta (Δ) efficiency is defined as the ratio of an increment in the external mechanical power output to the increase in metabolic power required to produce it. The purpose of the present study was to investigate whether differences in leg muscle activity between running and cycling can explain the observed difference in Δ efficiency between the two activities. A group of 11 subjects performed incremental submaximal running and cycling tests on successive days. The Δ efficiencies during running and cycling were based on five exercise stages. Electromyograph (EMG) measurements were made of three leg muscles (gastrocnemius, vastus lateralis and biceps femoris). Kendall's correlation coefficients between the mean EMG activity and the load applied were calculated for each muscle, for both running and cycling. As expected, the mean Δ efficiency during running (42%) was significantly greater than that during cycling (25%). For cycling, all muscles showed a significant correlation between mean EMG activity and the load applied. For running, however, only the gastrocnemius muscle showed a significant, but low correlation (r=0.33). The correlation coefficients of the vastus lateralis and biceps femoris muscles were not significantly different from 0. The results were interpreted as follows. In contrast to cycling, which includes only concentric contractions, during running up inclines eccentric muscle actions play an important role. With steeper inclines, more concentric contractions must be produced to overcome the external force, whereas the amount of eccentric muscle actions decreases. This change in the relative contribution of concentric and eccentric muscle actions, in combination with the fact that eccentric muscle actions require much less metabolic energy than concentric contractions, can explain the difference between the running and cycling Δ efficiency. Electronic Publication