Muscle fiber number in biceps brachii

in bodybuilders and control subjects

J. D. MAcDOUGALL, D. G. SALE, S. E. ALWAY, AND J. R. SUTTON

Departments of Physical Education and Medicine, McMaster University,
Hamilton, Ontario L8S 4Kl Canada

MACDOUGALL, J.D.,D. G. SALE, S.E. ALWAY, AND J.R. phy of existing fibers rather than by hyperplasia. The
SUTTON. Muscle fiber number in biceps brachii in bodybuilders training mode in this study, however, was of the endur-
and control subjects. J. Appl. Physiol.: Respirat. Environ. Ex- ance low-resistance type rather than the high-resistance
ercise Physiol. 57(5): 1399-1403, 1984.-Muscle fiber numbers training used in the studies that have reported hyperpla-
were estimated in vivo in biceps brachii in 5 elite male body- sia (8, 9).
builders, 7 intermediate caliber bodybuilders, and 13 age- Competitive bodybuilders possess extreme degrees of
matched controls. Mean fiber area and collagen volume density
muscle hypertrophy and thus are excellent models for

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were calculated from needle biopsies and muscle cross-sectional
area by computerized tomographic scanning. Contralateral investigating possible hyperplasia of fibers. We have
measurements in a subsample of seven subjects indicated the recently published data suggesting that elite bodybuilders
method for estimation of fiber numbers to have adequate reli- have more muscle fibers in triceps brachii than a group
ability. There was a wide interindividual range for fiber num- of trained control subjects (14). The present study was
bers in biceps (172,085---418,884), but despite large differences prompted by our wish to further examine this phenom-
in muscle size both bodybuilder groups possessed the same enon by utilizing a technique which would permit a more
number of muscle fibers as the group of untrained controls. direct calculation of actual fiber number.
Although there was a high correlation between average cross- Since it has been demonstrated that fibers in the biceps
sectional fiber area and total muscle cross-sectional area within extend from origin to insertion of the muscle (1, 2, 5),
each group, many of the subjects with the largest muscles also
tended to have a large number of fibers. Since there were equally measurement of muscle cross-sectional area at the belly
well-trained subjects with fewer than normal fiber numbers, we of the muscle includes all muscle fibers. Thus by use of
interpret this finding to be due to genetic endowment rather computerized tomographic (CT) scanning to determine
than to training-induced hyperplasia. The proportion of muscle muscle area and by calculation of average fiber area from
comprised of connective and other noncontractile tissue was needle biopsies, the total number of fibers may be esti-
the same for all subjects (-13%), thus indicating greater abso- mated.
lute amounts of connective tissue in the trained subjects. We Since biceps is a muscle that is trained to achieve
conclude that in humans, heavy resistance training directed maximal hypertrophy in bodybuilders, it is particularly
toward achieving maximum size in skeletal muscle does not suited for investigation of possible hyperplasia. It was
result in an increase in fiber numbers. our hypothesis that if heavy-resistance training induces
an increase in fiber number, the biceps of bodybuilders
computerized tomographic scanning; fiber hyperplasia; connec- who have trained intensively for 6-10 yr should show
tive tissue evidence of fiber hyperplasia in comparison with control
subjects of the same age.
ITHAS GENERALLYBEENACCEPTED sincel897(15)that METHODS
the fiber content of mammalian skeletal muscle remains
constant after birth and that muscle growth occurs ex- Needle biopsies were taken from the biceps brachii of
clusively through enlargement of the existing fibers. Re- the dominant arm of 12 male bodybuilders-5 of whom
cently, however, several studies (8, 9, 11, 16, 19) have were considered as elite and 7 of whom we classified as
suggested that compensatory and training-induced being of intermediate caliber. The elite group not only
growth in muscle may be the result of both hypertrophy had significantly greater muscle sizes than the others but
of existing fibers and an increase in fiber numbers. had also won at least one major physique contest. In
Gollnick and colleagues (7) have suggested that meth- addition, tissue was taken from 13 age-matched control
odological errors associated with estimation of fiber num- male subjects who had no previous history of resistance
ber from histological sections may have affected the training. The biopsies were taken from the muscle at the
results of these studies. Utilizing a technique by which estimated point of greatest girth. All subjects partici-
all of the fibers in a muscle were isolated and counted, pated with their own informed consent in accordance
these authors (7) concluded that in rats muscle enlarge- with the policies of the Ethics Committee of the Mc-
ment caused by ablation of a synergist and treadmill Master University Medical Centre.
running could be completely accounted for by hypertro- The tissue was sectioned on a cryostat and stained by
0161-7567/84 $1.50 Copyright 0 1984 the American Physiological Society 1399
1400 MAcDOUGALL, SALE, ALWAY, AND SUTTON

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FIG. 1. Computer tomographic scan of upper arm with elbow extended illustrating biceps bra&iii (BB).

means of a modified Gomori trichrome stain for myofi- cross-sectional muscle area
brillar adenosinetriphosphatase (following preincubation (corrected for connective tissue)
at several pHs) before being photographed under the numbers of fibers =
average fiber area
light microscope. Cross-sectional area for both type I and
type II fibers (as classified by the pH 10.0 preincubation) where
was measured by a computerized digitizer for a minimum average fiber area
of 200 fibers of each type per biopsy. Percent fiber type
was estimated by counting a minimum of 550 fibers/ (% type I)(mean type I area)
biopsy. The proportion of collagen and other noncon- -t (% type II) (mean type II area)
tractile tissue was calculated in the trichrome-stained =
100
sections by means of a 168 point, point-counting tech-
nique (20). and connective tissue correction equals the total muscle
Cross-sectional area of biceps was determined from area minus the percentage of muscle that is collagen and
CT scans (Ohio Nuclear, model 20-30) of the upper arm other noncontractile tissue.
with the elbow extended (Fig. 1). Several scans were To evaluate the reliability of this method for the
taken at l-cm intervals-one at the estimated point of estimation of fiber numbers, biopsies were also taken
greatest circumference for biceps and one proximal and from both arms of seven of the control subjects and fiber
distal to that point. The CT scan negatives were printed number estimated for both right and left arms.
to 17 X 22 in. prints and biceps area measured by
computerized digitizer. The scan yielding the greatest RESULTS
area was then selected.
Muscle fiber number was calculated as follows Reliability of fiber enumeration technique. Based on the
FIBER NUMBER IN BICEPS
assumption that there is an equal number of fibers in
the left and right biceps, the reliability of the technique
for estimating fiber numbers is illustrated in Fig. 2. The
correlation coefficient for fiber number for left and right
biceps for the seven control subjects was 0.79 (P < 0.01)
with an SEE of t 11%. Stated in other terms, the mean
intrasubject difference between right and left arms with
this technique was t 8.9% with a t 3.6% SD.
PhysicaL characteristics. The physical characteristics
of the three groups of subjects are presented in Table 1.
Both groups of bodybuilders had been training inten-
sively for approximately the same period (6-8 yr). The
elite group was significantly heavier (P < 0.001) and had
significantly larger biceps areas (P < 0.005) than both
the intermediate and control groups. The intermediate
bodybuilder group also had significantly larger biceps
areas (P < 0.005) than did the untrained control group.
Four of the five elite bodybuilders admitted to the use of
anabolic steroids, as did four of seven intermediate bod-
BRACHII 1401
groups. Average fiber areas for both bodybuilder groups
were significantly greater (P c 0.02) than for the control
group. Although the average fiber area of the elite group
appeared to be -13% larger than that of the intermediate
group, the difference was not statistically significant.
There was a very wide interindividual range in esti-
mated fiber numbers in all three groups. Fiber numbers
ranged from 172,000 to 381,000 among the control sub-
jects, from 198,000 to 374,000 among the intermediate
bodybuilders and from 204,000 to 419,000 among the
elite subjects. The mean results for each group, however,
did not differ (Table 1 and Fig. 3).
Connectiue and noncontractile tissue. The proportion
of connective and other noncontractile tissue was ex-
tremely consistent between individuals and was -13% of
the tissue sample. Of this, -6% was identified as collagen
and 7% as other tissue.

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DISCUSSION
ybuilders.
Fiber areas and fiber number. Biceps brachii was found The results of this study show that there are large
to be comprised of -61 t 5.1% type II fibers with no differences in fiber numbers in biceps brachii in human
differences in percent fiber type between any of the subjects and that despite extremely large differences in
muscle size, a group of bodybuilders possess the same
30 number of muscle fibers as a group of untrained controls.
/ Validity of fiber number estimation method. The valid-
rz5 0 /
x / ity of our method for estimating fiber number rests on
O / several assumptions. First, one must assume that the
e25 /
/ fibers in biceps extend the total length of the muscle.
0
8 / Although it has been shown anatomically (1) and elec-
zz / tromyographically (2) that some fibers extend from ori-
/
t20 gin to insertion of the muscle, it is not known whether
O/
!!I! / all fibers do. Nevertheless we feel that it is highly likely
0 that all fibers at least pass through the belly of the
2 0
%I5 / muscle which was the point at which the measurements
r=0.79
ii /
SEE=+ll%
were taken for the present study.
/
/
Second, since estimates of fiber number are based on
s
/ measurement of fiber cross-sectional area, it must be
frwl0 /
m /
accepted that any shrinkage or swelling of tissue during
t / histological preparation would bias results. However
z / since tissue preparation methods were constant for all
IL , ,, , , , , , , , , , , , , , , , , , , l subjects, such possible artifacts would not be considered
10 15 20 25 30 to affect interindividual comparison to the same extent
NUMBER OF FIBERS, RIGHT BICEPS, x IO4 that they would affect absolute estimation of fiber num-
FIG. 2. Estimated fiber numbers in right and left biceps brachii in bers.
7 control subjects. Dotted Line indicates line of identity. Third, since the needle biopsy technique results in
7 7 7 P p-7

1. Physical description, biceps area, average fiber area, ana esttmatea numoer
, l ,

TABLE OJ Jzoers
for elite and intermediate bodybuilders and for untrained controls

% Collagen No. of Fibers x lo3

Training Ht and Other Avg Fiber
Group Age, History, 9 m Biceps Area,
Noncon- Area x 102,
Yr cm kg mm2 Uncor-
tractile Pm2 Corrected
Yr rected
Tissue

Elite bodybuilders (n = 5) 23.7 6.2 178 90.3 2487" 13.0 105.6d 213.8 290.7
Al.2 to.7 Ik2.2 t2.9 t181 t3.6 k28.2 t53.2 k72.2
Intermediate bodybuilders (n = 7) 24.2 7.8 173 81.1" 1925b*C 12.1 93.7d 190.8 259.5
rt3.1 k4.4 t3.2 t4.0 k321 t5.4 k27.3 t51.3 t69.7
Untrained controls (n = 13) 22.5 0 179 78.2 1412b 13.4 67.0 204.8 278.5
to.5 k3.1 t2.3 t291 t2.3 k19.6 t44.6 k60.7
Values are means t SD. Fiber numbers are expressed both uncorrected and corrected for change in sarcomere. Length with needle biopsy
technique. a See text for explanation. bP -=I0.005. "P e 0.005. dP e 0.02. "P e 0.001.
1402 MAcDOUGALL, SALE, ALWAY, AND SUTTON

depending on the definition of resting length. Based upon

our CT scan differences in biceps area in the elbow
0 UNTRAINED CONTROLS extended and maximally flexed positions, we have arbi-
0 INTERMEDIATE BODYBUILDERS
trarily chosen a correction factor of 36%. Thus, assuming
o ELITE BODYBUILDERS
that the fiber volume remains constant regardless of
sarcomere length, we would estimate that the actual
number of muscle fibers exceeds our estimated number
by 36%.
Finally, one must assume that the biopsy sample which
is taken is representative of the belly of the muscle with
respect to fiber type distribution and muscle fiber areas.
r=0.71 (PcO.001) On the basis of recent studies (4,10,13) one must accept
that the biological nonhomogeneity of skeletal muscle
may contribute to a sampling error when only one biopsy
is used. We do not, however, feel that the magnitude of
such sampling errors would be sufficient to have a sys-
tematic nonrandom effect on our results. This assump-
tion is supported by our contralateral measurements.
Reliability. Since one would generally expect little var-

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iation in fiber number between left and right arms (6, 7),
we conclude that our method of estimating fiber numbers
is quite reliable (Fig. 2). We would also interpret the
slight differences which were found to be due to sampling
error rather than actual anatomical differences.
Muscle size and fiber numbers. When corrected for
sarcomere length, fiber numbers ranged from 172,085 to
418,884 fibers/muscle. These values are in agreement
with a previous autopsy study on human male subjects
(5). The relationship between muscle area and muscle
fiber number is shown in Fig. 3B. It can be seen that
r=0.35 (non significant)
although some of the elite bodybuilders possess a higher
-
than average number of fibers, there are a number of
untrained control subjects with just as many fibers.
I /I 1 I I I , II I1 I1 I 11 Our data indicate a high correlation between average
/5 1Q 15 20 25 30 35 40 cross-sectional fiber area and total muscle cross-sectional
AREA OF BICEPS BRACHII (mm2 x 102) area (Fig. 341) and confirm reports on human thigh
FIG. 3. A: correlation between cross-sectional area of biceps brachii muscle (17), and elbow flexor and extensor muscles (18)
and average fiber area (n = 25). B: correlation between cross-sectional in controls and bodybuilders. It is interesting to note
area of biceps brachii and estimated fiber number (n = 25). Although that although the muscle area of the elite body builders
statistically nonsignificant, it can be seen that larger muscle sizes are
partially result of greater muscle fiber numbers as well as greater fiber
is significantly larger than that of the intermediate
areas. group, fiber areas do not differ significantly between the
two groups. The massive arm size of the elite group is
fully contracted tissue, fiber areas were measured with thus partially the result of larger muscle fibers as well as
sarcomeres in the contracted state. Conversely, muscle being partially due to superior fiber numbers. Since the
area was measured by CT scanning with sarcomeres at intermediate group was equally trained, we do not inter-
resting length. It is thus apparent that the individual pret such differences to be due to training-induced hy-
fiber areas have been overestimated while the total num- perplasia but rather to genetic endowment. It is apparent
ber of fibers has been underestimated. Again, however, from the present data that wide variations occur in fiber
we conclude that this would be a constant error and numbers in biceps among the general population Al-
would not affect relative differences in fiber numbers though it is the size of the individual fibers that is the
between subjects. An alternative would be to obtain the main determinant of muscle size, it is reasonable to
CT scans with the elbow maximally flexed so that both expect the bodybuilder who has inherited a large number
measurements would be in the contracted state. This of fibers to have a greater potential than the bodybuilder
was also done in a number of subjects but we found that who inherits fewer fibers than normal. For this reason it
when the arm was in this position it was difficult to is probable that within a population of bodybuilders
separate the CT scan of biceps from brachialis muscle in those destined to achieve the greatest muscle size would
all subjects. In those in whom this was possible the biceps also tend to exhibit a large number of fibers. We consider
area was found to be -36 t 2% greater in the flexed than this to be a plausible explanation for our previous results
in the extended position. Estimates of the extent to (14) which suggested bodybuilders with extremely large
which a sarcomere shortens when fully contracted vary triceps have more fibers than a group of trained control
from 50-60% (3) to 30-40% of its resting length (12) subjects.
FIBER NUMBER IN BICEPS BRACHII 1403
Connective tissue. Our finding of a constant proportion
of colla .gen and other noncontractile tissue, regardless of In summary, this study indicates that large interindi-
muscle size or state of trainin , indicates that the abso- vidual differences in muscle fiber number occur in human
lute amount of connective tissue is considerably greater biceps and that highly trained bodybuilders do not pos-
in the bodybuilders than in the controls. This suggests sess more fibers than do untrained control subjects. We
that training-induced hypertrophy of muscle fibers is therefore conclude that heavy-resistance training di-
accompanied by a proportional increase in connective rected towards achieving maximum size in skeletal mus-
tissue. cles does not result in an increase in fiber numbers.
Anabolic steroids. In the present stu
dismiss the possibility that the use of anabolic steroids This study was supported by grants from the Muscular Dystrophy
by the bodybuilders may have affected fiber size. Our Association and the Natural Sciences and Engineering Research Coun-
findings, however, indicate that the use of steroids had cil of Canada.
no apparent effect on fiber number since both the elite We are grateful to Pauline McCullagh for technical assistance and
to Laura Diskin for secretarial assistance.
and intermediate groups of bodybuilders ha
roids and yet had the same number of fibers as the Received 6 July 1983; accepted in final form 13 June 1984.

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