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


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


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.


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.


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


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.

References

ASHMORE, C. R. and L. DOERR, 1971: Compara- tive aspects of muscle fiber types in different species. Exp. Neurol., 31: 408-418.

BEATTY, C. H., R. D. PETERSON and R. M. BOCEK, 1963: Metabolism of red and white muscle fiber groups. Am. J. Physiol., 204: 939-942.

BEATTY, C. H., G. M. BASINGER and R. BOCKER, 1967: Differentiation of red and white fibers

in muscle from fetal, neonatal and infant rhesus monkeys. J. Histochem. Cytochem., 15: 93-103.

BOCEK, R. M. and C. H. BEATTY, 1966: Glycogen synthetase and phosphorylase in red and white

muscle of rat and rhesus monkey. J. Histo- chem. Cytochem., 14: 549-559.

EDGERTON, V. R. and D. R. SIMPSON, 1969: The intermediate muscle fiber of rats and guinea

pigs. J. Histochem. Cytochem., 17: 828-838. HERBERT, Y., 1967: Neural regulation of enzymes

in muscle fibers of red and white muscle. Exp. Neurol., 19: 92-103.

ITO, R, S. INOKUCHI, T. KIMURA and M. KOzu, 1980: Myofibrous organization of M. trapezius

of crab-eating monkey. J. Anthrop. Soc. Nip-

pon., 88: 375-385 (in Japanese).[伊藤良作・猪口清一郎・木村忠直・神津正明,

1980:カニクイザル憎帽筋の筋線維構成について.

人類学雑誌,88:375-386.]

ITO, R., K. IHARA, S. ONDA, T. KIMURA, T. AJIRI, H. NAKANISHI and S. INOKUCHI, 1980:

Myofibrous organization of spinohumeral muscles of crab-eating monkey. Showa Med. J.,

40: 531-544 (in Japanese).

[伊藤良作・恩田聡・木村忠直・阿尻貞三・中西

弘・猪口清一郎,1980:カニクイザル棘腕諸筋の

筋線維構成について.昭和医学会雑誌,40:531-

544.]

JOHNSON, M. A., J. POLGAR, D. WEIGHTMAN and

D. APPLETON, 1973: Data on the distribution of fiber types in thirty-six human muscles. An

autopsy study. J. Neurol. Sci., 18: 111-129. KIKucHI, K., 1973: Histological study on the

effect of muscular training upon the medium muscle fibers in skeletal muscles. J. Physical Fitness Japan, 22: 17-25 (in Japanese).

[菊地邦雄,1973:中間筋線維に及ぼす筋トレニン

グの影響に関する組織学的研究.体力科学,22:

17-25.]

KIMURA, T., R. ITO, T. SATO and I. SATO, 1979: Myofibrous organization of M. tibialis anterior

in crab-eating monkey. Showa Med. J., 39: 381-388 (in Japanese).

[木村忠直・伊藤良作・佐藤亨・佐藤厳,1979:

カニクイザル前脛骨筋における筋線維構成につい

て.昭和医学会雑誌,39:381-388.]

KIMURA, T., 1980: Distribution pattern of muscle fiber type in M. tibialis anterior of crab- eating monkey. J. Anthrop. Soc. Nippon., 88:

183 (in Japanese).

[木村忠直,1980:カニクイザル前脛骨筋における

筋線維型の分布について.人類学雑誌,88:183.]

MORRIS, C. J., 1970: The significance of inter-

mediate fibers in reinnervated human skeletal

muscle. J. Neurol. Sci., 11: 129-136.

OGATA, T., 1958a: A histochemical study of the

red and white muscle fibers. Part I. Activity

of the succinoxydase system in muscle fibers.

Acta Med. Okayama, 12: 216-227.

OGATA, T., 1958b: A histochemical study of the

red and white muscle fibers. Part II. Activity

of the cytochrome oxidase in muscle fibers.

Acta Med. Okayama, 12: 228-232.

OXNARD, C. E., 1963: Locomotor adaptations in

the primate forelimb. Symp. Zool. Soc., London,

10: 165-182.

PETER, J. B., R. J. BARNARD, V. R. EDGERTON,

C. A. GILLESPIN and K. E. STEVIPEL, 1972:

Metabolic profiles of three fiber types of skeletal

muscle in Guinea pigs and rabbits. Biochemis-

try, 11: 2627-2633.

SICKLES, D. W. and C. A. PINKSTAFF, 1981: Com-

parative histochemical study of prosimian primate hindlimb muscles. II. Populations of fiber

types. Am. J. Anat., 160: 187-194.

STEIN, J. M. and H. A. PADYKULA, 1962: Histo-

chemical classification of individual skeletal

muscle fibers of the rat. Am. J. Anat., 110:

103-123.


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

distribution pattern of muscle fiber type in musculus

Documents