Distribution Pattern of Muscle Fiber Type in Musculus ...
Transcript of Distribution Pattern of Muscle Fiber Type in Musculus ...
人 類 誌 J.Anthrop. Soc.Nippon
93(3): 371-380 (1985)
Distribution Pattern of Muscle Fiber Type in
Musculus Biceps Brachii of White-Handed Gibbon
Tadanao KIMURA and Seiichiro INOKUCHI
Department of Anatomy, School of Medicine, Showa University
Abstract Histochemical examinations of muscle fiber types by Sudan black
B staining were made on m. biceps brachii of male and female white-handed
gibbon (Hylobates lar). Three types of muscle fibers could be discriminated in terms of the reaction to the pigment and the cellular diameter : red muscle
fiber (type I) with a positive reaction and a small diameter ; white muscle
fiber (type II) with a weak reaction and a large diameter ; intermediate muscle
fiber (type III) with an intermediate reaction and diameter. Of 4648 muscle
fiber cells, in average of male and female, scanned in the cross-section, red
muscle fibers accounted for 44.1%, white muscle fibers 28.3%, and inter-
mediate muscle fibers 27.6%, respectively. White muscle fibers were clearly
localized in the external layers of both heads of m. biceps brachii. In con-
trast, red muscle fibers were more evenly distributed throughout the muscle though moderately localized in the regions adjacent to the sulcus bicipitalis
medialis. Intermediate muscle fibers showed no specific localization at all.
These results suggest that m. biceps brachii of the gibbon, an acrobatic arm-
swinger, is inclined to be a red muscle that is fatigue-resisting rather than
power-generating.
Keywords Muscle fiber type, Locomotion, Gibbon, M. biceps brachii
Introduction
Bimanual suspensory locomotion is an ex-
traordinary positional behavior found exclu-
sively in primates, particularly in the anthro-
poid age. This mode of locomotion, in which the trunk of the animal is held up-
right, has been variously regarded by num-
erous investigators as related to the evolu-
tion of human bipedalism. Thus, a sub-
stantial number of studies have been made
on the musculoskeletal gross anatomy of the
upper extremities in the ape (e. g. OXNARD,
1963). Also partinent in this context, how-
ever, may be an analysis of the morpho-
physiological features of the muscle tissues. It has been well-established that there are
at least two types of fiber cells composing
the skeletal muscle : red muscle fibers (Type
I) involved in slow and tonic contraction
and white muscle fibers (Type II) involved
in fast and phasic contraction (OGATA,
Article No. 8505 Received March 27, 1985.
372 T. KIMURA and S. INOKUCHI
1958b; BEATTY et al., 1963; BOCEK &
BEATTY, 1966; BEATTY et al., 1967; HER-
BERT, 1967; ASHMORE & DAERR, 1971).
From the enzymohistochemical as well as
biophysical reaction of muscle fibers, the
presence of muscle fibers intermediate be-tween Type I and Type II has been also
revealed (EDGERTON et al., 1967; MORRIS,
1970; KIKUCHI, 1973). Using the lipoid
stain Sudan black B, by which the lipid
content of the myofibrous sarcoplasm can
be estimated, we have discriminated three
types of muscle fibers in the limb muscles
of the macaque (KIMURA et al., 1979; ITo
et al., 1979; ITo et al., 1980). As for the
apes, however, no attempt of analyzing
muscle fiber composition seems to have been
done so far. The present study was de-
signed to elucidate the myofibrous organiza-
tion of the upper limb muscles as an adapta-
tion in the ape to the bimanual suspensory
behavior. Accordingly, subject was the gib-
bon, a specialist of this acrobatic behavior,
and samples were collected from the biceps
brachii muscle, a elbow flexor that is likely to participate in the brachiation.
Materials and Methods
Materials were obtained from two adult
white-handed gibbons (Hylobates 1ar), one
male and one female (Table 1). The re-sected m. biceps brachii were fixed in Baker's
solution and washed with water. Both the
caput longum and caput breve were cut (10 mm thick) at the maximum venter. These
sections were embedded in 10% gelatin.
Lyophilized slices 20-25 * in thickness were
prepared with a Sartorius type of microsome, and immediately they were stained with Sudan black B according to Lison's method.
About tenfold magnified photographs of
Table 1. Data of the white handed gibbon.
the muscle tissue specimens were prepared.
Scanning lines were plotted along the sagit-
tal axis (Y-axis) and the transverse axis (x-
axis) along the polarity of m. biceps brachii.
The caput longum sections were divided
equally into external, intermediate, and in-
ternal parts, and the caput breve sections
into external and internal parts (Fig. 1).
Under a microscope, an ocular micrometer
of 100 * on each side was placed at the
intersection of the Y-axis and X-axis. The
muscle fibers were classified into three types
as described below. By means of the micro-
meter, the three types of muscle fibers
were scannned continuously along each line
by changing the position of a crusiately-
movable stage in microcope. In the deter-
mination, the cells outside of the lines in
the left side and in the upper side of the
micrometer were excluded from the count.
In contrast, the cells outside of the right-
hand and of the lower base lines were
counted. Thus, cells weae always counted
in the same way. A schematic presentation
of the examples is shown in Fig. 2.
Distribution Pattern of Muscle Fiber Types 373
Fig. 1. Cross-sectional area in M. Biceps brachii of white handed gibbon.
Results
Fig. 2. Scanning method of muscle fiber type by micrometer.
A-Line 11*Point : Number 11 of the sagittal axis (Y-axis) in caput longum of Fig. 1.
W: White muscle fiber. I : Intermediate muscle fiber.
R : Red muscle fiber. x : Non counting side.
Q : Counting side.
A) Historogical findings of muscular tissue
The cross-sectional features of the muscle
tissues stained with Sudan black B revealed
that muscle fibers with different diameters
reacted selectively to the pigment, enabling
a classification of the three types of muscle
fiber cells (Photo 1). Red muscle fibers
(Type I) with a small diameter particularly were found to show a highly positive reac-
tion. The sarcoplasm of the red muscle
fibers contains a large amount of lipid, and
it has been shown by electron microscopic
findings that many large mitochondria are
located in the sarcoplasm. The positive re-
action to the pigment of red muscle fibers
conceivably resulted from the synergistic
and selective reaction to the lipoid stain,
Sudan black B, of lipids in the sarcoplasm
374 T. KIMURA and S. INOKUCHI
Photo. 1. Cross-section in M. biceps branchii of the white handed gibbon
by Sudan Black B staining * 400.
S : red muscle fiber, I : Intermediate muscle fiber, W : white muscle fiber.
and phospholipids sticking to the cristae and
matrices of the mitochondria.
In contrast, white muscle fibers (Type II)
with a large diameter did not react to the
pigment. A very small amount of lipid is contained in the sarcoplasm of white muscle
fibers, while a large amount of glycogen is
contained in it. Unlike the red muscle
fibers, mitochondria in the cells are small
and few, resulting in the fact that white
muscle fibers showed the weakest reaction
to Sudan black B among the three types.
Intermediate muscle fibers (Type III) with
an intermediate diameter showed a reaction
to the pigment intermediate between red
and white muscle fibers. The type III muscle
fiber cells have been believed to have the
sarcoplasm of both Types I and II.
Longitudinal sections of m. biceps brachii
(Photo 2) also showed that each type of muscle fibers reacted differently to the pig-
ment in its own manner. In particular, the
white muscle fibers, that contain the small
amount of lipid, showed a definitely striated
structure.
Thus, three types of muscle fiber cells
with corresponding diameters, widely ob-
served in the skeletal muscle of mammals,
were clearly identified in m. biceps brachii
of the gibbon. In some cross-sectional speci-
mens, however, the diameters of muscle
fibers were not in accordance with the re-
action to the pigment. As shown in Photo
3, a highly positive reaction was observed
in some of the fibers with larger diameters.
They could be assigned to the red muscle
fibers from the standpoint of the reaction to
Sudan black B. On the other hand, the
muscle fibers reacting weakly to the pig-
ment, which therefore should have been
assigned to the white muscle fibers, had
smaller diameters. Occurrence of these
Distribution Pattern of Muscle Fiber Types 375
Photo. 2. Longitudinal section of the M. biceps brachii * 400.
For abbreviations, see Photo. 1.
Photo. 3. Three types of the muscle fiber by Sudan Black B * 400.
For abbreviations, see Photo. 1.
376 T. KIMURA and S. INOKUCHI
muscle fibers, however, was confined to
specimens located in the proximal region of
the sulcus bicipitalis medialis in m. biceps
brachii of the female gibbon. As for muscle
fibers in the other regions of the same
specimen, a general classification of the
fiber types was applicable.
B) Location and distribution of muscle
fiber types
Fig. 3 shows histograms of the cross-
sectional distribution of the muscle fiber
types. The frequency (°o) of each muscle
fiber types was plotted for each scanning
line of the tissue specimens. Since the
localization of fiber types was not observed
along the sagittal direction of the muscle,
only the transverse distribution of the types
is illustrated.
1) Frequency of white muscle fibers
For this type of muscle fiber, both sexes showed a high frequency in the external
part ; 31.7% (male) and 33.5% (female), on the average, for the caput longum, and
29.6% (male) and 35.3°o (female) for the
caput breve. Thus, the frequency was
slightly higher in the female than in the
male. The frequency in the intermediate
part of the caput longum was slightly lower than those in the external parts, without significant difference between the male
(30.8%) and the female (28.3%). Frequencies in the internal parts of both caputs were
lower by about 10% than those in the other
parts ; 20.4°0 (male) and 25.2% (female) for the caput longum, and 21.1% (male) and
24.7% (female) for the caput breve.
Since the proportion of the superficial layer
is largest in the external parts and least in
Fig. 3. Distribution pattern of muscle fiber type in M. biceps brachii
of the white handed gibbon.
Distribution Pattern of Muscle Fiber Types 377
the internal parts of m. biceps branchii, the
above results suggest that the frequency of
white muscle fibers in this muscle is highest
in the superficial layer and lowest in portions
adjacent to the sulcus bicipitalis medialis.
2) Frequency of intermediate muscle fibers
Although the frequency of intermediate
muscle fibers was somewhat low in the most
external portions of both caputs, no signi-
ficant difference in the frequency was indi-
cated between sites. The mean frequencies
for the caput longum were 28.7% (male) and
23.2% (female) in the external part, 28.5%
(male) and 25.4% (female) in the intermediate
part, and 25.8% (male) and 28.6% (female) in the internal part. Those for the caput
breve were 28.6% (male) and 26.0% (female)
in the external part and 29.4% (male) and
32.2% (female) in the internal part. In
general, the distribution frequencies were uniform, about 28%, among the sites, sug-
gesting no specific localization of this type of fibers.
3) Frequency of red muscle fibers
Among the three types of muscle fibers,
red muscle fiber shows the highest frequency.
For the caput longum, the mean frequencies
were 39.6% (male) and 43.4% (female) in
the external part, and 40.7% (male) and
46.4% (female) in the intermediate part, and
53.8% (male) and 46.2% (female) in the
internal part. Those for the caput breve
were 41.8% (male) and 38.6% (female) in
the external part and 49.6% (male) and 43.1%
(female) in the internal part. The frequency was thus higher by more than 15% than
that of white muscle fibers in each site ex-
cluding the most external portions where
the frequency of red muscle fibers was
slightly lower than that of white muscle
fibers. The frequency exceeded 50% parti-
cularly in the internal part of both caputs
of the male. Thus, localization of red
muscle fibers in both caputs increased in the
internal region adjacent to the sulcus bici-
pitalis medialis of m. biceps brachii, showing a distribution frequency that contrasted with
that of white muscle fibers.
C) Numbers of the three types of muscle
fiber
By means of a square ocular micrometer
of 100 *on each side, muscle fibers in the
cross-sectional area of m. biceps brachii
(mean area : 531.7 mm2) were scanned at intervals of 0.5 mm and 1 mm along the Y-
axis and the X-axis, respectively. As shown
in Table 2, the mean total number of the
three types of muscle fiber was 4648 cells.
The number of red muscle fibers (Type I)
was largest, 2049 about 44.1% of the total
number. Although the number of inter-
mediate muscle fibers (Type III) was slightly
larger than that of white muscle fibers (Type
II) in the male, and vice versa in the female,
there was no significant difference in the
mean frequency between Type II and III.
The above distribution frequencies suggests
that m. biceps brachii of the white-handed
gibbon is highly likely to be of the red muscle type concerned with the tonic con-
traction.
Table 2. Number and percentage of muscle
fiber type in M. biceps brachii of the
white banded gibbon.
378 T. KIMURA and S. INOKucHI
Discussion
There have been many reports on the dif-
ference in contraction mechanism and histo-
chemical specificity of the three types of
muscle fibers, whose contens have also been
clarified. The function of individual skeletal
muscles, terefore, can be determined histo-
chemically by correctly quantifying the loca-
tion and distribution of each type of muscle
fibers. It has already been reported by
JOHNSON (1973) and SICKLES et al. (1981) that white muscle fibers and red muscle
fibers have a tendency to be localized in the
superficial layer and deep layer, respectively,
of a muscle.
Enzymohistochemical classification, which
is very frequently used in recent years for
identification of muscle fiber types at the
cellular level, has a disadvantage : the
method cannot be applied to muscle tissues
which have been fixed in formalin for a
long time. In the present study, we adopted
the histochemical method employing the
lipoid stain, Sudan black B, which is com-
patible with the fixation of muscle tissue with formalin. This procedure is based on
the fact that each type of muscle fiber cell
is selectively stained with the pigment ac-
cording to the difference in the lipid content
of its sarcoplasm. The red, white, and
intermediate muscle fibers classified by the
present method with Sudan black B corre-spond to the slow-twitch-oxidative, f ast-
twitch-oxidative-glycolytic and fast-twitch-
glycolytic fibers, respectively, as suggested by PETER et al. (1972). OGATA (1958a) and
STEIN et al. (1962) recognized that the
muscle fiber type classification by Sudan
black B staining corresponds to that by the
activity of succinic dehydrogenase, an oxida-
tive enzyme. This phenomenon seems to be
the result of a different quantity of mito-
chondria contained in each type of the
muscle fiber, as mentioned above. Sudan
black B staining method thus seems adequate
for classification of the muscle fiber types.
For the examination of the location and
distribution of the muscle fiber types, it is
most important to scan and quantify an ap-
propriate part according to the shape and anatomical positional relations of the skeletal
muscle. KIMURA (1980) scanned the caput,
venter, and cauda of the cross-sectional
areas of m. tibialis anterior in the crabeating
macaque (Macaca f ascicularis) along the
polarity. The muscle fibers in these sites showed a similar tendency without any signi-
ficant difference in the distribution frequency
among the sites. On the basis of this ob-
servation, the present authors scanned the
cross-sectional tissues at the maximum ven-
ter of m. biceps brachii of the white-handed
gibbon, and found that red muscle fibers is prevailing in this muscle. As far as the authors are aware, this is the first analysis
of the muscle fiber types in the anthropoid
ape.
The significance of the above observation
lies in the fact that m. biceps brachii of the
gibbon, despite its positive role assumed in the arm-swinging, tends to be a red, rather
than a white, muscle. The observation sug-
gests that a high power is not required for the elbow flexors in the hylobatid brachia-
tion, and that a fatigue-resisting rather than
power-generating property of these muscles is critical for the bimanual suspensory be-
havior. Red muscle fibers larger in diameter
than white muscle fibers, shown to exist in
the present materials, seem adaptive in the
above context. Whether or not, however,
such a feature is a morpho-physiological
adaptation of the muscle to the bimanual
Distribution Pattern of Muscle Fiber Types 379
suspensory behavior is not clear at this time,
since muscle fibers with 'inversed' histo-
chemical attributes was localized to limited
regions of the muscle.
This study was supported by Grant-in-Aid
for Cooperative Research No. 57340051
(Senior Researcher : Morihiko Okada) from The Ministry of Education, Science and
Culture.
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D. APPLETON, 1973: Data on the distribution of fiber types in thirty-six human muscles. An
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380 T. KIMURA and S. INOKUCHI
和 文 抄 録
シ ロテテナ ガザルの上腕二頭筋 におけ る筋 線維型の分布について
木 村 忠 直 ・猪 口 清 一 郎
昭和大学医学部解剖学教室
類人猿の懸垂行動への適応形態に関する分析
は,筋 骨格系における肉眼解剖学的な比較研究
に よるものが多 くなされているが,筋 収縮の最
小単位である筋線維細胞の組織化学的な観点か
らの接近は,今 だ試みられていない。そこで本
研究では,シ ロテテナガザル (Hylobates lar)
雌雄各 一頭における上腕二頭筋の極性にそった
筋腹横断面の筋線維細胞を Sudan Black B の
染色法によって,機 能性を異にする三タイプの
筋線維細胞を分類 し,そ の分布頻度や局在性よ
り,シ ロテテナガザルにおける上腕二頭筋の機
能形態的な内容を追求 した。 Scanning 法によ
って算出した筋線維数は4648細 胞(雌 雄の平均
値)で,そ のうち持続的な緊張性収縮を示すタ
イプIの 赤筋線維が全体の41.1%で 最も分布
頻度が高かった。特に内側の上腕二頭筋溝近位
に移行するに伴って比率が高 くなる傾向を示し
た。これに対 し,瞬 発力に富み相動性収縮を示
すタイプIIの 白筋線維は28.3%で,皮 膚側の
浅層部において分布頻度が高かった。次いで,
わずかの差でタイプIとIIの 両形質を兼そなえ
ているタイプIIIの中間筋線維は26.7%と な り,
筋の極性にそって均等な分布頻度を示し局在性
は認められなかった。以上の筋線維構成の結果
より,シ ロテテナガザルにおける上腕二頭筋は,
持続的な緊張性収縮 (tonic contraction)を 示
す赤筋型の傾向を有 している骨格筋であ り,懸
垂行動への機能形態的な適応性が示唆された。
木 村 忠 直 昭和大学医学部解剖学教室
〒142 東京都品川区旗の台1-5-8
Tadanao KIMURA Department of Anatomy, School of Medicine, Showa University
1-5-8, Hatanodai, Shinagawa-ku, Tokyo 142, Japan