Scand J Med Sci Sports 2003: 13: 4047 Printed in Denmark All rights reserved

COPYRIGHT BLACKWELL MUNKSGAARD 2003 ISSN 0905-7188

S C A N D I N A V I A N

MEDICINE & SCIENCE IN SPORTS

J O U R N A L

O F

Muscle fibre type adaptation in the elderly human muscle

Jesper L. Andersen
Copenhagen Muscle Research Center, Department of Molecular Muscle Biology, Rigshospitalet, Copenhagen, Denmark Corresponding author: Jesper L. Andersen, Ph D, Copenhagen Muscle Research Center, Department of Molecular Muscle Biology, Rigshospitalet, Section 9312 Juliane Mariesvej 20, 1th DK-2100, Copenhagen , Denmark Accepted for publication 29 August 2002

This short review discusses changes in the fibre type distribution, myosin heavy chain isoform composition and histological appearance of the very elderly human skeletal muscle. Point of origin of the discussion comes from data that we have obtained from muscle biopsies from the vastus lateralis muscle of a group of frail very elderly subjects (age: 88 6 3 years, range 8597). Myosin heavy chain composition of muscle homogenates and single fibres, fibre type distribution, fibre size and capillary density were examined and compared with muscle biopsies from the young vastus lateralis muscle. Histological preparations of the muscle biopsies from our elderly subjects showed extended grouping (Nygaard & Sanchez, Anat Rec 1992: 202: 451459) of the fibre types as well as significant changes in the appearance and size of the individual muscle fibres.

On average, the fibre type composition of our very elderly subjects do not seem to be different to what is observed in a corresponding young group when examined with ATPase histochemistry. Likewise, the MHC composition of the muscle homogenates is comparable to what is observed in young subjects. Nevertheless, a detailed examination of the MHC composition of single fibres from the old subjects revealed that the most prominent phenotype was fibres coexpressing MHC I and MHC IIA. This is very different from what is observed in the young muscle. Detailed investigation of longitudinally cut fibres indicated that some fibres in the very old muscle, in contrast to the young muscle, switch fibre type along the length of the fibre or contain areas or nuclear domains in which the MHC expression is different from the remaining part of the fibre.

Discussion The proportions of the human skeletal muscle change, as we get older. Pronounced muscle atrophy, muscle weakening and an apparent slowing are inevitable followers of increasing age. These changes have significant implications on the individual and his or hers ability to overcome everyday tasks (Porter, Vandervoort, Lexell, 1995). The present review aim at giving an overview of the morphological and biochemical fibre type related modifications that occurs with increased ageing as they were manifested in a small population of very elderly subjects that we have investigated. We analysed needle biopsies obtained from the vastus lateralis muscle of a group (n 12) of frail very elderly men and women (age: 88 + 3 years, range 8597) (Andersen, Terzis, Kryger, 1999). The muscle biopsies were very thoroughly examined in order to establish fibre type distribution, fibre size and myosin heavy chain (MHC) composition in muscle homogenates and single fibres, as well as density of capillaries. Furthermore, the biopsies were, in a qualitative manner, examined for morphological appearance and abnormalities as they appeared in comparison with the young vastus lateralis muscle.

Muscle morphology
In a number of studies examining either whole muscle preparations or muscle biopsies of elderly subjects an apparent grouping of the muscle fibre types have been observed (Nygaard & Sanchez, 1982; Lexell, Downham, Sjostrom, 1986; Lexell & Downham, 1991). Grouping describes the phenomenon that certain fibre types seem to be distributed in clusters rather than in the random fashion commonly observed in younger muscle. The grouping phenomenon was quite pronounced in our group of frail elderly (fig. 1). Almost all individuals showed some sign of fibre type grouping, although the range was considerable, ranging from biopsies showing a normal random appearance of the fibre types, to biopsies with very pronounced clustering of the fibre types (fig. 1). The mechanisms for the obvious grouping of the fibre types in the ageing human muscle still need to be fully understood. Presently, the most evident explanation seems to be that the fibre type grouping arises from a continuous process of denervation and partial reinnervation that is claimed to accelerate with advancing age (for review see Lexell, 1995). Apart from the grouping phenomenon other morphological features separate the old from the young muscle.

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Fibres cut transversely will in the young muscle of course have different shapes but almost all fibres (in the vastus lateralis muscle) will have a certain angular shape. In the very elderly muscle many fibres will appear flattened, crushed or even bananashaped (fig. 2). This flattening of the fibres is definitely more pronounced among the type II than the type I fibres. The reason for this change in appearance and architecture of the fibres are essentially unknown. It could be an intrinsic ageing process or even the very first sign of a programmed cell death. More activityrelated reasons could also be the cause. The fact that the type II fibres are more exposed than the type I fibres could be viewed as a reflection of age-related type II disuse. This would fit well with the fact that very elderly people are not subjected to force and power demanding muscle-work, and thus do not or very seldom fully activate specific portions of the type II fibres, ergo they slowly wither and atrophy due to lack of stimulation. If the latter is the main explanation, at least something can be done to retard and delay the onset of these changes.

Fibre size
An obvious consequence of ageing is a marked atrophy of the skeletal muscles. This atrophy is also evident in the individual muscle fibres (Larsson, Sjodin, Karlsson, 1978; Larsson, Grimby, Karlsson, 1979; Scelsi, Marchetti, Poggi, 1980; Larsson, 1982; Lexell, Taylor, Sjostrom, 1988; Klitgaard, Mantoni, Schiaffino et al., 1990a; Lexell, 1995; Borges & Essen-Gustavsson, 1989; Essen-Gustavsson & Borges, 1986). Nevertheless, the age-related fibre atrophy is very inhomogeneous when looking at the fibre pool as a whole. In muscle biopsies from very elderly subjects it

Young muscle

Old muscle

Fig. 1. Grouping of muscle fibre types in the ageing muscle. ATPase staining of a young muscle (age: 22 years) and old (87 years) muscle. Note the random distribution of the fibre types in the young muscle vs. the pronounced grouping of the fibre types in the old muscle. Dark fibres, type I fibres; White fibres, type IIA fibres; Grey fibres, type IIX (or I/IIA) fibres; Bar 100 mm. ATPase staining (pH 4.6).

Young muscle

Old muscle

Fig. 2. Fibre shapes are often different in young and very old human skeletal muscle. Muscle fibres in the young muscle most often appear angular with four to six angels or corners, whereas many fibres in the elderly muscle appear as if they have been flattened or crushed. This flattening of the fibres is much more pronounced among the type II fibres than the type I fibres. Dark fibres, type I fibres; White fibres, type IIA fibres; Grey fibres, type IIX (or I/IIA) fibres; Bar 50 mm. ATPase staining (pH 4.6).

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can often be observed that in some areas the fibres have almost normal size, and right next to these are areas in which the fibres are small, or even extremely small (fig. 3). Furthermore, the atrophy also affects the different fibre types differently. Numerous studies have demonstrated that ageing-related fibre atrophy is less hard on type I fibres than on type II fibres (for review see Porter et al., 1995). When we compared our 88 year old subjects with a matching group of 25 year old subjects, we found that the type I fibres were reduced in size, but only to 75% of the young type I fibres (2891 + 853 mm2 vs. 3822 + 755 mm2). In contrast, the type II fibres were reduced to 43% of the young fibre size (1704 + 936 mm2 vs. 3974 + 873 mm2) (fig. 4). taken into account the difference between the young and the old muscle is erased. Thus, the old muscle will contain equal amounts of capillaries per square millimetre (Coggan, Spina, King et al., 1992a, b). Using the same protocol (Qu, Andersen, Zhou, 1997) we analyzed capillary density of both a young group of subjects (Qu et al., 1997) and of our group of very elderly subjects. When calculated as capillaries/fibre we found that the young subjects had more than twice the number of capillaries per fibre compared to the elderly group, but since the younger group as mentioned earlier have larger fibres this may not be the correct way to compare the two groups. Thus, we also looked at the number of capillaries per square millimetre in the two groups. If calculated in this manner it seems that the very old subjects per area have the same, or even more, capillaries than the young group (536 vs. 474 capillaries/mm2) (Qu et al., 1997; Andersen & Kryger, unpublished).

Capillary density
Most studies have indicated no severe age-related influence on skeletal muscle capillary density (Cartee, 1994). If measured as capillaries/fibre the ageing muscle will show a decrease in capillary density (Frontera, Meredith, O'Reilly, Evans, 1990; Cartee, 1994), but if the reduced fibre size of the elderly muscle fibres are

Fibre types
An often-debated issue is whether or not fibre type composition of the skeletal muscle change with ageing.

Young muscle

Old muscle

Fig. 3. Muscle fibre sections from a normal young muscle containing fibres of normal size and a section from a very old subject (> 85 years) in which areas with extremely atrophied fibres occurred. Bar 50 mm. ATPase staining (pH 4.6).

Type I fibre size 6000 5000 4000 m2 3000 2000 1000 0 0 20 40 60 Age (years) 80 100 6000 5000 4000 m2 3000 2000 1000 0 0 20

Type II fibre size

40 60 Age (years)

80

100

Fig. 4. Size of type I and type II fibres in a group of young (avg. age 25 years) and old subjects (avg. age 88 years). Notice the apparent lager difference between the young and the old group in size of the type II fibres in comparison to the type I fibres (Andersen & Kryger, unpublished).

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Several studies have addressed this question either directly or indirectly (Scelsi et al., 1980; Nygaard & Sanchez, 1982; Larsson, 1983; Lexell, Henriksson m, Larsen, Winblad, Sjostro 1983; Grimby, Aniansson, Zetterberg, Saltin, 1984; Aniansson, Hedberg, Henning, Grimby, 1986; Lexell et al., 1986; Klitgaard et al., 1990a; Chilibeck, Paterson, Cunningham, Taylor, Noble, 1997; Hakkinen, Newton, Gordon et al., 1998). The answer to the question is ambiguous. A few studies comparing groups of young and old subjects have reached conclusions that point in the direction of a change in fibre type composition in the aged muscle, generally more type I fibre has been found in the aged muscle (Larsson et al., 1978; Larsson, 1983; Jakobsson, Borg, Edstrom, Grimby, 1988). Nevertheless, the overwhelming number of studies has found no significant different fibre type composition in the aged muscle (for review see Porter et al., 1995 Lexell, 1995). Very few truly longitudinal studies have been conducted, but quite recently a study with a biopsy interval of 12 years were published (Frontera, Hughes, Fielding, Fiatarone, Evans, Roubenoff, 2000). In this study a group of elderly subjects were biopsied at the age of 65 years and again at the age of 77 years. At age 65, they had 60% type I fibres, but at age 77 they had only 40% type I fibres. This longitudinal study seems to contradict what has been suggested in a substantial number of earlier cross-sectional studies in the sense that it shows a decrease in the number of type I fibres with ageing. The cross-sectional studies prompting the idea of a slightly increased number of type I fibres with ageing have compared a young group of subjects with a group of elderly subjects in their late sixties or early seventies (Larsson et al., 1978; Larsson, 1983; Jakobsson et al., 1988). Cross-sectional studies examining fibre type distribution in subjects in their late eighties and nineties hardly exists, but do not indicate an increased amount of type I fibres (Andersen et al., 1999; Tomonaga, 1977; Scelsi et al., 1980). Thus, perhaps the amount of type I fibres do not increase with ageing as a general rule, but it could be that a window exists around the age of 60 or 70 years in which there is a provisional slight increase in the number of type I fibres. If we imagine that this is actually so, how do we then explain this? It is known that the number of functioning motor units decreases with age (McComas, 1996). Furthermore, this process of degeneration of the motor units seems to start accelerating around the age of 60 years. If it assume that the majority of the first motor units die are large motor units innervating type II fibres, and that the reinnervation of the fibres are only partial and relative randomly, this would create a situation favouring an increased relative amount of type I fibres. If then, later in life (in the late seventies and onwards) the motor units supplying the type I fibres also degenerate and die, then the balance between the type I and the type II fibres would be re-established in the very elderly muscle, leaving a bump on the type I curve around the age of 6070 years. At least our group of subjects in their late eighties show no indication of an increased relative amount of type I fibres, if anything they show the contrary. Truly longitudinal studies expanding over 40 or 50 years would throw new light over this problem, but will for obvious reasons probably never be conducted.

Co-expression of MHC isoforms

In an attempt to perform a detailed description of the fibre type composition of our group of very elderly subjects, we decided to dissect out single fibres from the muscle biopsies and subsequently determine the MHC isoform composition of the single fibres by gelelectrophoresis (Andersen et al., 1999). We dissected out 2264 single muscle fibres from the 12 biopsies. This analysis revealed a number of interesting facts. We found that only 20% of the fibres contained only MHC I. This is about less than half found in younger muscle (Klitgaard, Bergman, Betto et al., 1990b; Andersen, Klitgaard, Saltin, 1994a; Andersen, Klitgaard, Bangsbo, Saltin, 1994b; Zhou, Klitgaard, Saltin, Roy, Edgerton, Gollnick, 1995). We found that 27% of the fibres contained only MHC IIA and 22% co-expressed both MHC IIA and IIX, which is about the same number as seen in the young muscle (Andersen et al., 1994a, b; Klitgaard et al., 1990b; Zhou et al., 1995). Only very few fibres (0.3%) contained only MHC IIX, which is a low number, but not an uncommon observation in young muscle (Andersen et al., 1994a, b; Klitgaard et al., 1990b; Zhou et al., 1995; Larsson, Li, Berg, Frontera, 1996). As a novel finding we observed that a few fibres had a very unusual combination of MHC isoforms. Thus, 1.2% of the fibres contained all three isoforms (MHC I, IIA and IIX) and 0.7% of the fibres contained MHC I and MHC IIX, without any MHC IIA. Both of these phenotype has not been registered in normal young vastus lateralis before (Andersen et al., 1999). Furthermore, and also somewhat surprising we observed that the most common fibre phenotype was fibres co-expressing MHC I and MHC IIA (28.5%). This should be compared to the small numbers found in younger muscle (15%) (Klitgaard et al., 1990b; Andersen et al., 1994a, b; Zhou et al., 1995; Larsson et al., 1996; Andersen et al., 1999). Additionally, when we looked closer at these fibres we observed that the majority of those fibres were fibres that were dominated by MHC I with only a minor fraction of MHC IIA (fig. 5b). This was in contrast to the fibres co-expressing MHC IIA and MHC IIX, in which the majority of the fibres expressed equal amounts of the two isoforms (fig. 5c). When we examined our ATPase stainings, we found much less fibres expressing both MHC I and MHC IIA (these fibres are often referred to as IIC fibres in

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(a)
40 35 30 25 Percent 20 15 10 5 0 MHC I MHC I/IIA MHC IIA MHC IIA/IIX MHC IIX MHC I/IIX MHC I/IIA/IIX

(b)
80 70 60 Percent

(c)
80 70 60 Percent 50 40 30 20 10 0
MHC I/IIA (dominant I) MHC I/IIA MHC I/IIA (no dominance) (dominant IIA)

50 40 30 20 10 0

MHC IIA/IIX MHC IIA/IIX MHC IIA/IIX (dominant IIA) (no dominance) (dominant IIX)

Fig. 5. (a) MHC composition (%) of single muscle fibres from the vastus lateralis muscle of a group of very old subjects (avg. age 88 years) (Andersen et al., 1999); (b) Single fibres co-expressing MHC I and MHC IIA, subdivided into groups according to the dominant MHC isoform; (c) Single fibres co-expressing MHC IIA and MHC IIX, subdivided into groups according to the dominant MHC isoform.

histochemical terminology) than expected from the single fiber data. This apparent divergence in results, generated from the different methods of analysis, provided a problem. One obvious difference between the two methods is that with ATPase histochemistry only very narrow part of the fibre (10 mm thick) is used in the analysis, whereas the single fibres used for the gelelectrophoretic determination of the MHC isoforms are often several millimetre long, thus the amount of muscle fibre examined is often several hundred fold longer. Traditionally, mixed or hybrid fibres are often looked upon as having a fairly even distribution of the two MHC isoforms expressed throughout the entire length of the fibre, which can to some extent be verified by the fact that a type I/IIA (IIC) fibres can be followed in serial sections of good quality biopsies (Andersen, unpublished observations). Animal experiments using chronic electrical stimulation, have shown that at least under certain conditions the MHC isoform distribution might not be so uniform along the length of the fibre (Staron & Pette, 1987). Skeletal muscle fibres are multinucleated and a large number of nuclei are distributed along the length of the individual fibres. At least for the contractile proteins, the individual nuclei have some sort of autonomy of the surrounding area, so that transcribed mRNA is translated into protein and incorporated locally in the domain. However, under normal conditions and certainly in the young muscle there seems to be a strict coordination of the expression of mRNA for the various MHC isoforms, so that all the nuclei behave alike. A condition for this type of organization is the before

mentioned strict coordination of the individual nuclei within the fibre. Consequently, the individual nucleus within the fibre transcribes the same mRNA of the MHC isoforms or the same two MHC mRNAs in the same ratio. This mechanism will ensure that a type I fibre stays a type I fibre and a type II fibre stays a type II fibre. Furthermore, when a certain stimuli are induced this mechanism ensures that a fibre type shift happens throughout the entire fibre. What happens if for some reason this coordination is lost or is reeling, if the individual nuclei looses the coordination and steps out, either alone or in clusters? If this would happen, nuclear domains or parts of fibres would probably transcribe mRNA and additional locally incorporate a MHC isoform that could be different from the rest of the fibre or different from areas close to. It is not difficult to imagine that at very high age this coordination could be disturbed and some or many nuclei could start misbehaving either for inbuilt intrinsic or other reasons. This scenario could in part explain why we found only a minor part of the fibres to be type I/IIA fibres with ATPase staining, using only a 10-mm crosssection of the fibres representing only one or two nuclei domains. In contrast, the several millimetre long fibre pieces used in the single fibre analysis representing several hundred nuclear domains. Although we have not systematically cut in the length direction of the fibres, cross-sectional cutting often have part of the biopsy cut longitudinally. Thus, we re-evaluated our ATPase stainings in an attempt to find evidence of fibres with different MHC expression along the length of the fibres. Interestingly, we found several examples

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of fibres with a different ATPase staining pattern along the length of the fibre as well as examples of smalldemarcated areas within the individual fibres showing a different staining pattern from the rest of the fibre. Latter, these small areas were in size comparable to the nuclear domain of a single nucleus. Examples of the two types of findings are shown in figs 6 and 7. This type of phenomenon is never seen in non-diseased young muscle. If this phenomenon of fibre type shifts along the length of the fiber and individual or clusters of nuclei expressing another type of MHC than the remaining part of the fibre is common in the very elderly muscle, this could in part explain the divergence between the methods of analysing. The reason why the very elderly muscle fibres apparently looses coordination control remains to be uncovered. In conclusion, the fundamental question whether human skeletal muscle fibre type/MHC isoform composition changes with age can probably not be answered with a simple yes or no. Furthermore, our data from the analysis of single muscle fibres shows that there is not so much a change in the ratio between type I and type II fibres in the classic sense but an obfuscation of the border between type I and type II fibres, so that close to one third of the fibres in the fibre pool at very old age are neither strictly type I nor strictly type II fibres but fibres co-expressing both MHC I and IIA (Andersen et al., 1999).

Fig. 6. Longitudinally cut muscle fibre originating from the muscle of a very old subject (> 85 years). The fibre apparently shifts phenotype along the length-axis of the fibre. Below is shown a theoretical sketch of how different transcription of mRNA for the MHC isoforms on each side of the shifting-zone might provide a fibre type shift along the length-axis of the fibre. For further information see text. ATPase staining (pH 4.3).

= Muscle nucleus = MHC IIA mRNA = MHC I mRNA

Fig. 7. Longitudinally cut muscle fibre originating from the muscle of a very old subject (> 85 years). Notice how the staining pattern changes in a small area within the fibre. Below is shown a theoretical sketch of how different transcription of mRNA for the MHC isoforms in a small area corresponding to the domain of a single nucleus could provide a very local area expressing a different MHC isoform than the remaining part of the fibre. For further information see text. ATPase staining (pH 4.3).

= Muscle nucleus = MHC IIA mRNA = MHC I mRNA

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Perspectives Ageing causes muscle fibre loss and fibre atrophy, progressively leading towards a state at which an elderly person will be unable to conduct most every-day tasks. Thus, it is of the outmost importance that studies examining the most effective ways of strengthening the elderly muscle as well as preventing fibre atrophy or even introducing hypertrophy are initiated. I am convinced that the most effective and simple advice to give to the elderly population, in an attempt to counteract agerelated muscle degeneration, is to prescribe regular resistance training. Our finding of extremely high relative amount of fibres co-expressing MHC I and MHC IIA in the very elderly muscle are currently difficult to link to any clinical significance at the whole muscle level. Nevertheless, it will be interesting to pursue our hypothesis of local shift in MHC expression along the length of the individual muscle fibres, in an attempt to establish if this apparently age related phenomenon can be liked to changes in the expression of various factors controlling fibre type specificity. Key words: ageing; capillary density; co-expression; fibre size; MHC isoforms; myosin; nuclear domain. Acknowledgement
The author would like to acknowledge the skilful work on the single fibre project done by Greasimos Terzis. Likewise, Ann Kryger worked hard on obtaining the muscle biopsies from this very unique group of subjects. The Danish National Research Foundation supported this work, grant number 50414.

References
Andersen JL, Klitgaard H, Bangsbo J, Saltin B. Myosin heavy chain isoforms in single fibres from m. vastus lateralis of soccer players: effects of strength-training. Acta Physiol Scand 1994b: 150: 2126. Andersen JL, Klitgaard H, Saltin B. Myosin heavy chain isoforms in single fibres from m. vastus lateralis of sprinters: influence of training. Acta Physiol Scand 1994a: 151: 135142. Andersen JL, Terzis G, Kryger A. Increase in the degree of co-expression of myosin heavy chain isoforms in skeletal muscle fibres of the very old. Muscle Nerve 1999: 22: 449454. Aniansson A, Hedberg M, Henning GB, Grimby G. Muscle morphology, enzymatic activity, and muscle strength in elderly men: a follow-up study. Muscle Nerve 1986: 9: 585591. Borges O, Essen-Gustavsson B. Enzyme activities in type I and type II fibres of human skeletal muscle in relation to age and torque development. Acta Physiol Scand 1989: 136: 2936. Cartee GD. Ageing skeletal muscle: Response to exercise. Exe Sport Sci Rev 1994: 22: 91120. Chilibeck PD, Paterson DH, Cunningham DA, Taylor AW, Noble EG. Muscle capillarization O2 diffusion distance, and VO2 kinetics in old and young individuals. J Appl Physiol 1997: 82: 6369. Coggan AR, Spina RJ, King DS, et al. Histochemical and enzymatic comparison of the gastrocnemius muscle of young and elderly men and women. J Gerontol 1992a: 47: B71B76. Coggan AR, Spina RJ, King DS, et al. Skeletal muscle adaptations to endurance training in 60 to 70 yr old men and women. J Appl Physiol 1992b: 72: 17801786. Essen-Gustavsson B, Borges O. Histochemical and metabolic characteristics of human skeletal muscle in relation to age. Acta Physiol Scand 1986: 126: 107114. Frontera WR, Hughes VA, Fielding RA, Fiatarone MA, Evans WJ, Roubenoff R. Aging of skeletal muscle: a 12 yr longitudinal study. J Appl Physiol 2000: 88: 13211326. Frontera WR, Meredith CN, O'Reilly KP, Evans WJ. Strength training and determinants of Vo2max on older men. J Appl Physiol 1990: 68: 329333. Grimby G, Aniansson A, Zetterberg C, Saltin B. Is there a change in relative muscle fibre composition with age? Clin Physiol 1984: 4: 189194. Hakkinen K, Newton RU, Gordon SE, et al. Changes in muscle morphology, electromyographic activity, and force production characteristics during progressive strength training in young and older men. J Gerontol 1998: 53: B415B423. Jakobsson F, Borg K, Edstrom L, Grimby L. Use of motor units in relation to muscle fibre type and size in man. Muscle Nerve 1988: 11: 12111218. Klitgaard H, Bergman O, Betto R, et al. Co-Existence of myosin heavy chain I and IIa isoforms in human skeletal muscle fibres with endurance training. Pflugers Arch 1990b: 416: 470472. Klitgaard H, Mantoni M, Schiaffino S, et al. Function, morphology and protein expression of ageing skeletal muscle: a cross-sectional study of elderly men with different training backgrounds. Acta Physiol Scand 1990a: 140: 4154. Larsson L, Grimby G, Karlsson J. Muscle strength and speed of movement in relation to age and muscle morphology. J Appl Physiol 1979: 46: 451456. Larsson L. Histochemical characteristics of human skeletal muscle during aging. Acta Physiol Scand 1983: 117: 469471. Larsson L, Li X, Berg HE, Frontera WR. Effects of removal of weight-bearing function on contractibility and myosin isoform composition in single human skeletal muscle cells. Pflugers Arch 1996: 432: 320328. Larsson L. Physical training effects on muscle morphology in sedentary males at different ages. Med Sci Sports Exerc 1982: 14: 203206. Larsson L, Sjodin B, Karlsson J. Histochemical and biochemical change in human skeletal muscle with age in sedentary males, age 2265 years. Acta Physiol Scand 1978: 103: 3139. Lexell J, Downham D, Sjostrom M. Distribution of different fibre types in human skeletal muscles. Fibre type arrangement in m. vastus lateralis from three groups of healthy men between 15 and 83 years. J Neurol Sci 1986: 72: 211222. Lexell J, Downham DY. The occurrence of fibre-type grouping in healthy human muscle: a quantitative study of cross-sections of whole vastus lateralis from men between 15 and 83 years. Acta Neuropathol 1991: 81: 377381. Lexell J, Henriksson-Larsen, Winblad B, Sjostrom M. Distribution of different fiber types in human skeletal muscles: effects of aging studied in whole muscle cross sections. Muscle Nerve 1983: 6: 588595. Lexell J. Human aging, muscle mass, and fibre type composition. J Gerontol 1995: 50: 1116. Lexell J, Taylor CC, Sjostrom M. What is the cause of ageing atrophy? Total number, size and proportion of different

46

Muscle fibre type adaptation

fibre types studied in whole vastus lateralis muscle from 15 to 83 year old men. J Neurol Sci 1988: 84: 275294. McComas AJ. Skeletal Muscle: Form and Function. Chap. 22. 1996, Champaign, IL: Human Kinetics. Nygaard E, Sanchez J. Intramuscular variation of fibre types in the brachial biceps and the lateral vastus muscle of elderly men: how representative is a small biopsy sample? Anat Rec 1982: 203: 451459. Porter MM, Vandervoort AA, Lexell J. Aging of human muscle: structure, function and adaptability. Med Sci Sports Exerc 1995: 5: 129142. Qu Z, Andersen JL, Zhou S. Visualisation of capillaries in human skeletal muscle. Histochem Cell Biol 1997: 107: 169174. Scelsi R, Marchetti C, Poggi P. Histochemical and ultrastructural aspects of m. vastus lateralis in sedentary old people (age 6589 years). Acta Neuropathol 1980: 51: 99105. Staron RS, Pette D. Nonuniform myosin expression along single fibres of chronically stimulated and contralateral rabbit tibialis anterior muscles. Pflugers Arch 1987: 409: 6773. Tomonaga M. Histochemical and ultrastructural changes in senile human skeletal muscle. J Am Geriatr Soc 1977: 25: 125131. Zhou M, Klitgaard H, Saltin B, Roy RR, Edgerton VR, Gollnick PD. Myosin heavy chain isoforms of human muscle after short-term spaceflight. J Appl Physiol 1995: 78: 17401744.

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