Adaptations in skeletal strength training

muscle following

D. L. COSTILL, E. F. COYLE, W. F. FINK, G. R. LESMES, AND F. A. WITZMANN Human Performance Laboratory, Ball State University,

Muncie,

Indiana

47306

COSTILL, D. L., E. F. COYLE, W. F. FINK, G. R. LESMES, AND F. A. WITZMANN. Adaptations in skeZetaZ muscle following strength training. J. Appl. Physiol.: Respirat. Environ. Exercise Physiol. 46(l): 96-99, 1979. -Five men were studied before and after 7 wk of isokinetic strength training to determine its effects on muscle enzyme activities and fiber composition. One of the subject’s legs was trained using 10 repeated 6-s maximal work bouts, while the other leg performed repeated 30-s maximal knee extension exercise. The total work accomplished by each leg was constant. Training 4 times/wk achieved similar gains in peak torque for both legs at the training velocity (3.14 rad/s) and at slower speeds. Fatigability of the knee extensor muscles, as measured by a 60-s exercise test, was similar in both legs after training. Biopsy specimens showed significant changes in the % of the muscle area composed of type I and IIa fibers as a result of both strength training programs. In terms of muscle enzymes, only the 30 s exercise program resulted in elevated glycolytic, ATP-CP and mitochondrial activities. Despite these changes, none of the parameters measured were found to be related to the gains in either muscle strength or fatigability during maximal isokinetic contractions. muscle enzymes; muscle histochemistry; muscle fatigue

ADAPTATIONS

IN SKELETAL MUSCLE following endurance training are well documented (6, 10) and demonstrate an enhanced capacity for oxidative metabolism. Adjustments to strength training, on the other hand, are not so well defined. The intent of the present investigation is to assess the effects of two modes of maximal isokinetic training on muscle enzyme activities and fiber composition. A comparison has been made between training programs that required 6- and 30-s bouts of maximal isokinetic contractions of the knee extensor muscles. The muscle enzymes selected for study are representative of the metabolic systems responsible for energy derivation during anaerobic (i.e., creatine phosphokinase, myokinase, phosphorylase, phosphofructokinase, and lactate dehydrogenase) and aerobic (i.e., succinate dehydrogenase and malate dehydrogenase) muscular effort.

METHODS

Five healthy males served as subjects in this investigation. The age, height and weight for the men averaged (tSE)23.6 t 1.3 yr, 177.2 t 2.6 cm, and77.3 t 4.7 kg, respectively. All subjects were fully informed of the
96

risks and stresses associated with this research before giving their written consent to participate. Measurements. Testing and training of the knee extensor muscles were conducted with the aid of an isokinetic dynamometer (Cybex II, Lumex, Inc.) as previously described by Thorstensson (17). Before and after training, each of the subject’s legs was tested for peak torque at knee extension velocities ranging from 1.05 to 5.23 rad/s. In addition, an anaerobic performance test was done with each leg. This test consisted of maximal knee flexions and extensions at 3.14 rad/s for 60 s. Work output was summed every 10 s during this test. Four biopsies were obtained from the vastus lateralis muscle of each leg before and after training. One portion of the specimen was positioned in a mounting medium and frozen in isopentane cooled to the temperature of liquid nitrogen for histochemical analysis. The remaining biopsy material was weighed, frozen in liquid nitrogen and later assayed for muscle enzyme activities. The myofibrillar ATPase method (4, 16) was used for muscle fiber classification. Tissue slices (10 pm thick) were preincubated at a pH 4.3 and 4.6, and the staining reaction carried out at a pH 9.4. Muscle fibers were subsequently classified as types I (slow twitch oxidative), IIa (fast twitch, oxidative), and IIb (fast twitch, glycolytic). Fiber areas were determined from serial cross sections of each muscle sample after staining for NADH-diaphorase (15). The mean muscle fiber area for each sample was determined by averaging the crosssectional areas of 20 fibers that had been photographically reproduced and measured by planimetry. Enzyme anaZyses. Muscle samples analyzed for succinate dehydrogenase (SDH), total (cytoplasmic plus mitochondrial) malate dehydrogenase (MDH), lactate dehydrogenase (LDH), and total phosphorylase were first homogenized at 4°C in an 0.1 M triethanolamine (TEA) buffer (pH 7.6; 0.5% bovine serum albumin (BSA) 5 mM 2-mercaptoethanol). We have noted that the use of this buffer results in SDH activity that is 0.5 times that obtained with a PO, homogenizing buffer. The proportionality of the SDH activity produced with the TEA and PO, buffers is constant and does not alter the interpretation of the findings in the present study. The reaction media and fluorometric methods for these enzyme assays have been detailed earlier (3, 11). Homogenates for phosphofructokinase (PFK) were prepared in 100 mM potassium phosphate buffer (pH
Copyright 0 1979 the American Physiological Society

0161-7567/79/0000-0000$01.25

MUSCLE

ENZYME

ACTIVITIES

FOLLOWING

STRENGTH

TRAINING 300

97

8.2), containing 10 mM glutathione, 0.5 mM ATP, 5 mM MgSOd, and 30 mM NaF (1). The fluorometric determination of NADH disappearance was performed at 25°C using a reaction cocktail described by Mansour et al. (12) Creatine phosphokinase (CPK) and myokinase (MK) activities were determined fluorometrically on whole muscle homogenized in a TEA-BSA buffer (see SDH method above), containing 10 mM cysteine or 1.0 mM glutathione. The MK reaction was performed in a cocktail containing ADP (0.2 mM), MgCl, (5 mM), glucose (2 mM), NADP + (0.1 mM), 5 pg/ml hexokinase, and 1 pg/ml glucose-6-phosphate dehydrogenase. After determining the MK reaction rate, creatine phosphate (CP) was added to the mixture to measure the activity of CPK (11). Training. The subjects trained each leg 4 times/wk for a period of 7 wk. One leg (6 s) was trained using 10 repeated 6-s maximal knee extensions, while the other leg (30 s) performed repeated 30-s maximal knee extension exercise. The rest intervals between the 6- and 30-s bouts was 114 s and 20 min, respectively. The 6-s leg was exercised first during each training session, and the work bouts for the 30-s leg were repeated until the total work performed was equal to that produced by the 6-s leg. All training bouts were executed at a velocity of 3.14 rad/s (18O”/s). The rationale for selecting training bouts of 6 and 30 s was based on our efforts to stress both systems of anaerobic ATP generation (ATP-CP and glycolytic systems). In preliminary studies we noted little or no muscle lactate increase following 6 s of maximal isokinetic exercise (unpublished observations). The 30-s maximal work, however, increased muscle lactate from a resting level of 0.9 to 19.4 mmol/kg wet wt. These data suggest that the 6 s of maximal exercise was achieved principally at the expense of the ATP-CP systems, while the 30 s bout stressed both the ATP-CP and glycolytic systems. Differences between means were tested for significance with a t test for paired observations.
RESULTS

250

TIME

(sec.)

1. Mean power performance test.
FIG. TABLE 1. Muscle and after training

values

for each 10 s of maximal

anaerobic

fiber composition

be fore

6-s Leg Pre Post Pre

30-s

Leg Post

%Fiber I IIa IIb

type 44.8 +5.3 30.0 t5.1 25.2 +2.3 pm2 6,106 +541 7,370 + 834 6,766 +614 41.6 t7.1 32.8 +6.5 25.6 k3.3 0.79 0.62 1.28 5,887 -+527 7,318 t700 7,116 2650 36.6* 26.3 37.2* +1.3 26.0 +2.7 1.02* 0.71 1.43* 4,865 1~381 6,233 + 563 5,773 ?419 42.6 k4.5 32.6 t5.0 24.8 k3.0 0.77 0.58 1.31 4,959 +283 7,592 *645 5,628 t715 34.8* k4.9 44.6* k4.6 20.2 k1.5 1.28* 0.58 2.21* 41.8 k4.7 34.8 t3.3 23.4 22.4 48.2 k5.3 28.4 k4.1 23.4 k3.3 46.8 +4.2 32.0 23.5 21.2 +2.4

Fiber areas, I IIa IIb %Fiber area

As a result of training, the subjects showed a significant (P < 0.05) increase in peak torque for both legs (6 and 30 s) at the velocity used during training (3.14 rad/ s) and at slower speeds. Improvements in peak torque ranged from a high of 14% (0 rad/s) to a low of 4% at 3.14 rad/s. No differences, however, were found between the strength gains of the 6-s and 30-s legs. Figure 1 illustrates that during the 60-s maximal exercise test, both the 6-s and 30-s legs increased (P < 0.05) their mean power output as a result of training. This apparent training advantage was noted only during the first 30 s of the test. Thereafter, the slope of this “fatigue” curve and the mean power values were the same in both legs before and after training. Mean (t SE) muscle fiber characteristics measured before and after training are presented in Table 1. Statistically, the percentage of type I, Ha, and IIb fibers did not change with either form of training. When computed as the nercentage of the muscle’s cross-sec-

IIa IIb %Fiber area ratio IIa/I IIb/I IIa/IIb

Values are means + SE for legs exercised with repeated bouts of 6- and 30-s maximal isokinetic contractions. %Fiber area represents the fraction of the total area of the muscle section that was composed of each fiber type. * Significant (P < 0.05) differences between means of pre- and posttraining.

tional area represented by each fiber type, however, the type I fibers showed a decrease (P < 0.05) of 5.0 and 7.8% in the 6-s and 30-s legs, respectively. At the same time, the type IIa fibers in both legs increased significantly (P < 0.05). The type IIb fibers remained unchanged with training. Neither the 6-s nor 30-s training

98
TABLE 2. Muscle enzyme activities before and after training

COSTILL

ET AL.

Phosphorylase Phosphofructokinase Lactate dehydrogenase Creatine phosphokinase Myokinase Malate dehydrogenase Succinate dehydrogenase

18.3 k2.0 64.5 +3.2 321 +32 611 +44 309 +18 44.0 +2.3 4.0 kO.7

17.4 +1.6 69.1* +2.8 329 +28 603 +47 308 *14 46.0 22.5 4.1 +0.6
l

18.1 22.0 61.2 +2.4 295 +26 609 k46 309 +14 47.1 k1.4 4.5 +0.6

19.6* a.9 74.6* k2.8 318 +22 702* AZ53 350* +5 53.7* 22.6 5.0* +0.6

Values are means + SE, in pmollg min, for legs exercised with repeated 6-and 30-s maximal isokinetic contractions. * Significant (P < 0.05) difference between means of pre- and posttraining.

programs had any effect on the cross sectional areas of any of the fiber types (I, IIa, IIb). Muscles from the 6-s-trained leg showed little change in enzyme activities (Table 2). Only muscle PFK activ(P < 0.05). The 30 s exercise ity increased significantly program, on the other hand, resulted in elevated (P < 0.05) glycolytic, ATP-CP, and Krebs cycle enzyme activities (Table 2). Of the muscle enzymes measured, only LDH activity failed to increase with the 30-s training program.
DISCUSSION

There are 3 major findings of this research. The first is that two 30-s maximal exercise bouts/day (4 days/wk) are sufficient stimulus to increase the activities of muscle phosphorylase, PFK, CPK, MK, MDH, and SDH. When the same quantity of work, however, was divided into repeated 6-s exercise bouts, only PFK activity increased. Thus, with the exception of PFK activity, the stimulus responsible for increasing muscle enzyme activities seems to be related to the duration of each maximal exercise bout rather than the quantity of work performed by the muscle. The second major observation in this investigation was that, while the muscle enzyme activities increased in the 30-s leg, the muscle’s resistance to fatigue was the same as the 6-s-trained leg which showed no increase in muscle enzymes (Fig. 1 and Table 2). Although an increase in glycolytic and ATP-CP enzymes is generally interpreted to reflect an enhanced anaerobic potential, the present data demonstrate that fatigue during repeated maximal contractions is independent of the muscle’s capacity to generate ATP via anaerobic metabolism. Despite the greater strength exhibited in the early seconds of the 60 s anaerobic performance test, mean power declined so rapidly that the trained legs exhibited the same mean power as the untrained legs after 30-40 s of effort (Fig. 1). One possible explanation for this greater rate of decline in the trained muscles is a more rapid depletion of muscle ATP and CP as a

imposed bY result of the incre ased en .ergy requirements the hi .gher power output in the trained muscles duri % the first half of the performance test. Recent studies (13), however, have demonstrated an increase in muscle ATP and CP concentrations with strength training, which could compensate for the greater energy demands at these higher power outputs. Another possible explanation for the more rapid decline in me-an power by the trained muscles might be attributed to the influence of pH on maximal muscular force. Hill (9) in 1940 demonstrated that the formation of lactic acid in response to stimulation stopped when muscle pH dropped to roughly 6.3. More recently Hermansen and Osnes (8) have measured the pH in exercising human skeletal muscle and have reported values as low as 6.35 at exhaustion. Because lowering cellular pH is known to slow the rate of glycolysis, it might be argued that the decrease in muscle pH which accompanied the anaerobic performance test could have limited ATP synthesis by reducing the effectiveness of the glycolytic enzymes. If this were the case, then one might expect to see a lesser rate of fatigue in the 30-strained leg, which shows the greatest enzymatic potential for glycolysis. The fact that both the 30- and 6-s trained legs showed similar rates of fatigue suggests that the factor responsible for muscular exhaustion may be independent of ATP resynthesis. Fuchs et al. (5) have suggested that increased hydrogen ion concentration in muscle reduces the binding of the fibrillar capacity for Ca2+ through an inactivation studies by Herprotein, troponin. Recent unpublished mansen (personal communication) have demonstrated that in skinned rabbit muscle fibers, a decrease in muscle pH results in a marked loss of contractile tension that persists despite the availability of ATP. Thus, the loss of mean power illustrated in Fig. 1 may be due, in Pad 9to the effects of increasing hydrogen ion concentration on the muscle’s contractile mechanism, rather than on the glycolytic ATP resynthesis, per se. It is interesting to note that during the final 30 s of and the anaerobic performance test bo th the trained untrained muscles produced the same mean power values. If we adhere to the concept that a decrease in pH was responsible for the decline in mean power, then we might speculate that its influence was greatest in the pH sensitive fast twitch (type II) muscle fibers (2). Thus, the rapid fatigue noted in the first 30 s of the test after training may be due to a decline in tension generated by the type II fibers. The mean power produced in the final 30 s of the test both before and after training may simply reflect the tension and lesser fatigability of the slow twitch (type I) fibers. This concept is supported by fatigue and muscle glycogen depletion studies conducted during repeated fast voluntary contractions in man (14, 19). These studies demonstrated that the decline in peak torque which accompanied repeated maximal contractions was positively correlated (r = 0.86) with the percentage of type II fibers in the contracting muscle. The third principal finding of this research was that the biopsy specimens showed a significant (P < 0.05) change in the fiber composition of the vastus lateralis

MUSCLE

ENZYME

ACTIVITIES

FOLLOWING

STRENGTH

TRAINING

99

muscle after 7 wk of strength training (Table 1). There was an increase in the IIa to I and IIa to IIb fiber area ratios of both the 30- and 6-s-trained muscles. Since neither the percentage nor cross sectional areas of the type I, IIa, and IIb fibers show statistical changes with training, caution should be used in speculating in the significance of the changes in fiber area ratios. It should, however, be noted that Thorstensson (18) also observed a selective hypertrophy of the fast twitch (type II) fibers after 8 wk of maximal isokinetic training. The changes seen in our muscle fiber composition (fiber area ratios) with strength training may be due to an increase in type IIa fibers, a shift in type I and IIb fibers to type IIa fibers, or merely chance error of the measurement. It is, however, interesting to note a recent study by Gonyea et al. (7) which clearly demonstrated muscle fiber splitting in the forelimb of the cat after strength training.
REFERENCES 1. BALDWIN, K. M., W. W. WINDER, R. L. TERJUNG, AND J. 0. HOLLOSZY. Glycolytic enzymes in different types of skeletal muscle: adaptations to exercise. Am. J. Physiol. 225: 962-966, 1973. 2. BURKE, R. E., AND V. R. EDGERTON. Motor unit properties and selective involvement in movement. In: Exercise and Sports Science Reviews, edited by J. H. Wilmore. New York: Academic, 1975, vol. 3, p. 31-81. 3. COSTILL, D. L., W. J. FINK, AND M. L. POLLOCK. Muscle fiber composition and enzyme activities of elite distance runners. Med. Sci. Sports. 8: 96-100, 1976. 4. DUBOWITZ, V., AND M. H. BROOKE. MuscZe Biopsy: A Modern Approach. Philadelphia, PA: Saunders, 1973, p. 51-61. 5. FUCHS, F., Y. REDDY, AND F. N. BRIGGS. The interaction of cations with the calcium-binding site of troponin. Biochim. Biophys. Acta. 221: 407-409, 1970. 6. GOLLNICK, P. D., AND L. HERMANSEN. Biochemical adaptations to exercise: anaerobic metabolism. In: Exercise and Sports Sciences Reviews, edited by J. H. Wilmore. New York: Academic 1973, vol. 1, p. l-43. 7. GONYEA, W., G. C. ERICSON, AND F. BONDE-PETERSEN. Skeletal muscle fiber splitting induced by weight-lifting exercise in cats. Acta Physiol. &and. 99: 105-109, 1977. 8. HERMANSEN, L., AND J. B. OSNES. Blood and muscle pH after maximal exercise in man. J. AppZ. PhysioZ. 32: 304-308, 1972. 9. HILL, D. K. Anaerobic recovery of heat. J. Physiol London. 98: 460-467, 1940. 10. HOLLOSZY, J. 0. Biochemical adaptations to exercise: aerobic

These studies of strength training demonstrate significant enzymatic changes and modifications in the composition (fiber area ratios) of skeletal muscle. Despite these adaptations, none of the parameters measured was found to be related to either muscle strength or fatigability during maximal isokinetic contractions. These data suggest that fatigue in maximal muscular effort is not dependent on the muscle’s anaerobic potential as measured by glycolytic and ATP-CP enzyme activities.
This research was supported by National Institutes of Health Grant HL-20408; Lumex Inc.; and the Ball State University Research Committee Grant Res. 79-1976. Current address: E. F. Coyle, Dept. of Physical Education, University of Arizona, Tucson, AZ 85724; G. R. Lesmes, Dept. of Physical Education, Northeastern Illinois State University, Chicago, IL 60625 Received 4 April 1978; accepted in final form 8 August 1978.

11 12

13

14 15

16 17. 18. 19.

metabolism. In: Exercise and Sports Sciences Reviews, edited by J. H. Wilmore. New York: Academic, 1973, vol. 1, p. 45-71. LOWRY, 0. H., AND J. V. PASSONNEAU. A FZexibZe System of’ Enzymatic Analysis. New York: Academic, 1972. MANSOUR, T. E., N. WAKID, AND H. M. SPROUSE. Studies on heart phosphofructokinase: purification, crystallization and properties of sheep heart phosphofructokinase. J. BioZ. Chem. 241: 1512-1521, 1966. MCDOUGALL, G. R. WARD, D. G. SALE, AND J. R. SUTTON. Biochemical adaptation of human skeletal muscle to heavy resistance training and immobilization. J. AppZ. PhysioZ.: Respirat. Environ. Exercise Physiol. 43: 700-703, 1977. NILSSON, J., P. TESCH, AND A. THORSTENSSON. Fatigue and EMG of repeated fast voluntary contractions in man. Acta PhysioZ. &and. 101: 194-198, 1977. NOVIKOFF, A. B., W. SHIN, AND J. DRUCKER. Mitochondrial localization of oxidation enzymes: staining results with two tetrazolium salts. J. Biophys. Biochem Cytol. 9: 47-61, 1961. PADYKULA, H. A., AND E. HERMAN. The specificity of the histochemical method for adenosine triphosphatase. J. Histothem. Cytochem. 3: 170-195, 1955. THORSTENSSON, A. Muscle strength, fiber types and enzyme activities in man. Acta Physiol. Stand. SuppZ. 443: 45, 1976. THORSTENSSON, A. Observations on strength training and detraining. Acta Physiol. Stand. 100: 491-493, 1977. and fiber THORSTENSSON, A., AND J. KARLSSON. Fatigability composition of human skeletal muscle. Acta Physiol. Stand. 98: 318-322. 1976.