Per-Olof Eriksson

Regenerative potential of human skeletal muscle during aging

In this study, we have investigated the consequences of aging on the regenerative capacity of human skeletal muscle by evaluating two parameters: (i) variation in telomere length which was used to evaluate the in vivo turn‐over and (ii) the proportion of satellite cells calculated as compared to the total number of nuclei in a muscle fibre. Two skeletal muscles which have different types of innervation were analysed: the biceps brachii, a limb muscle, and the masseter, a masticatory muscle. The biopsies were obtained from two groups: young adults (23 ± 1.15 years old) and aged adults (74 ± 4.25 years old). Our results showed that during adult life, minimum telomere lengths and mean telomere lengths remained stable in the two muscles. The mean number of myonuclei per fibre was lower in the biceps brachii than in the masseter but no significant change was observed in either muscle with increasing age. However, the number of satellite cells, expressed as a proportion of myonuclei, decreased with age in both muscles. Therefore, normal aging of skeletal muscle in vivo is reflected by the number of satellite cells available for regeneration, but not by the mean number of myonuclei per fibre or by telomere lengths. We conclude that a decrease in regenerative capacity with age may be partially explained by a reduced availability of satellite cells.

Histochemical and morphological muscle-fibre characteristics of the human masseter, the medial pterygoid and the temporal muscles

An extensive histochemical and quantitative analysis of various portions of the human masseter, the medial pterygoid and the temporal muscles was performed in young adult males with normal intermaxillary relationships and complete dentition. There was marked and locally radical intramuscular variability in the muscle-fibre composition. Each muscle and the subunits of the muscles exhibited a characteristic fibre pattern--both the relative frequency and the diameter of the various fibre types differed significantly between the different portions. The fibre pattern was quantitatively different to that of the human lateral pterygoid muscle and both quantitatively and qualitatively dissimilar to that of the human digastric muscle and that of normal limb and trunk muscles. A large proportion of the fibres were ATPase intermediate fibres and must be regarded as being a part of the normal fibre population of the human mandibular elevator muscles. Type IIA fibres were rare. As muscle-fibre differentiation is considered to be influenced by motoneurone function, it can be assumed that the complex fibre pattern of the jaw-closing muscles is related to the unique function of the human mandibular locomotor system. A functional specialization is suggested providing optimal jaw control. Compared with the lateral pterygoid muscle (with predominantly type I fibres) and the digastric muscle (with predominantly type II fibres), the heterogeneous fibre composition of the jaw-closing muscles probably reflects their more complicated activity pattern and functional requirements. The marked difference between the type I and the type II fibre diameters, type II fibres generally being smaller, might reflect evolutionary changes in the masticatory habits, such as adaptation to refined and soft food. The individual variability in fibre composition suggests various levels of utilization and varying ability to adapt to jaw-muscle hyperactivity, to resist fatigue.

Differences in myosin composition between human oro-facial, masticatory and limb muscles: enzyme-, immunohisto-and biochemical studies

SummaryImmunohistochemistry was used to determine the myosin composition of defined fibre types of three embryologically different adult muscles, the oro-facial, masseter and limb muscles. In addition, the myosin composition in whole muscle specimens was analysed with biochemical methods. Both similarities and differences between muscles in the content of myosin heavy chains and myosin light chains were found. Nevertheless, each muscle had its own distinct identity. Our results indicated the presence of a previously undetected fast myosin heavy chain isoform in the oro-facial type II fibre population, tentatively termed ‘fast F’. The masseter contained aberrant myosin isoforms, such as foetal myosin heavy chain and α-cardiac myosin heavy chain and unique combinations of myosin heavy chain isoforms which were not found in the limb or oro-facial muscles. The type IM and IIC fibres coexpressed slow and fast A myosin heavy chains in the oro-facial and limb muscles but slow and a fast B like myosin heavy chain in the masseter. While single oro-facial and limb muscle fibres contained one or two myosin heavy chain types, single masseter fibres coexpressed up to four different myosin heavy chain isoforms. Describing the fibres according to their expression of myosin heavy chain isozymes, up to five fibre types could be distinguished in the oro-facial and limb muscles and eight in the masseter. Oro-facial and limb muscles expressed five myosin light chains, MLC1S, MLC2S, MLC1F, MLC2F and MLC3F, and the masseter four, MLC1S, MLC2S, MLC1F, and, in addition, an embryonic myosin light chain, MLCtemb, which is usually not present in normal adult skeletal muscle. These results probably reflect the way the muscles have evolved to meet the specialized functional requirements imposed upon them and are in agreement with the previously proposed concept that jaw and limb muscles belong to two distinct allotypes.

Fibre Composition of Human Intrinsic Tongue Muscles

The muscle fibre composition of three human intrinsic tongue muscles, the longitudinalis, verticalis and transversus, was investigated in four anterior to posterior regions of the tongue using morphological and enzyme- and immunohistochemical techniques. All three muscles typically contained type I, IIA and IM/IIC fibres. Type I fibres expressed slow myosin heavy chain (MyHC), type II fibres fast MyHC, mainly fast A MyHC, whereas type IM/IIC coexpressed slow and fast MyHCs. Type II fibres were in the majority (60%), but regional differences in proportion and diameter of fibre types were obvious. The anterior region of the tongue contained a predominance of relatively small type II fibres (71%), in contrast to the posterior region which instead showed a majority of larger type I and type IM/IIC fibres (66%). In general, the fibre diameter was larger in the posterior region. This muscle fibre composition of the tongue differs from those of limb, orofacial and masticatory muscles, probably reflecting genotypic as well as phenotypic functional specialization in oral function. The predominance of type II fibres and the regional differences in fibre composition, together with intricate muscle structure, suggest generally fast and flexible actions in positioning and shaping the tongue, during vital tasks such as mastication, swallowing, respiration and speech.

Adverse changes in fibre type composition of the human masseter versus biceps brachii muscle during aging.

The fibre composition of functionally different regions, the superficial and the deep portions, of the human masseter and the biceps brachii muscles of six elderly subjects (mean age 74 years) was studied by morphological and enzyme-histochemical methods. When compared to previous data for young adults, the masseter muscle from elderly subjects showed a significant decrease in the proportion of type I fibres and an increase of type IM and II fibre types, concomitant with muscle fibre atrophy. In the old biceps, there were no changes in the type I fibre proportion and fibre diameter, but a significant decrease in the type IIB fibre content. The present results of changes in fibre composition, in addition to previous findings of functional deterioration of the old masseter, probably reflect a combination of age related degenerative and adaptive processes within the trigeminal neuromuscular system. When compared with previous findings in young adult masseter, it can be concluded that the human masseter muscle differs from limb muscles both in fibre composition and in structural response of the muscle during aging. These differences between the masseter and the biceps brachii muscle suggest muscle specific alterations during aging probably due to differences in genetical control, functional performance and nerve and hormonal influences.

Enzyme-histochemical and morphological characteristics of muscle fibre types in the human buccinator and orbicularis oris.

Human masticatory muscles, originating from the first branchial arch and innervated by the trigeminal nerve, have a fibre composition distinct from that of limb and trunk muscles. The zygomatic muscles, originating from the second branchial arch and innervated by the facial nerve, differ in fibre composition from either the masticatory or the limb and trunk muscles. To elucidate further the structural basis for function, and the influence of embryological origin and innervation on oro-facial muscles, the buccinator and orbicularis oris muscles, which originate from the second branchial arch and are innervated by the facial nerve, were investigated. Like the masticatory and zygomatic muscles, they have a large representation in the cerebral cortex. Both muscles were composed of type I, type IIA and a few type IIC fibres of about equal diameter. However, the type I fibres had a different myofibrillar ATPase reaction from those in masticatory, zygomatic, limb and trunk muscles; this was a moderate to strong staining at pH 9.4, indicating a special isomyosin composition. Whereas the buccinator was composed of 53% type I fibres, the orbicularis oris had a 71% predominance of type II fibres. In both muscles, the mean fibre diameter and its marked intramuscular variability were similar to earlier findings in the zygomatic muscles. No muscle spindles were found. The large number of type I fibres in the buccinator implies a capacity for endurance during continuous work at relatively low levels of force. The predominance of type II fibres in the orbicularis oris indicates that it is built up of fast-twitch motor units, related to properties such as rapid acceleration and high speed during intermittent oro-facial movements. The similarities and differences in fibre-type composition between the facial, masticatory and limb muscles imply that specific functional demands are of greater importance for muscle differentiation than embryological origin and nerve supply.

The reliability of histochemical fibre typing of human necropsy muscles

SummaryThe reliability of muscle fibre typing of post mortem specimens was investigated with special reference to the influence of time and temperature. In specimens stored at +4° C, muscle fibre typing could be reliably performed up to at least ten and fifteen days post mortem for the masseter and biceps brachii muscles respectively. The corresponding figures for storage at room temperature were three and six days. The difference in the preservation of enzyme activity between masticatory and limb muscles might be related to the demonstrated difference in the fibre type composition and thus the enzyme content and energy sources.

Opposite changes in myosin heavy chain composition of human masseter and biceps brachii muscles during aging

The myosin heavy chain (MyHC) content in functionally different parts of the human masseter muscle of six elderly and five young adult subjects (mean age 74 and 22 years, respectively) was determined, using gel electrophoresis. The MyHC composition of the old masseter was also studied by enzyme- and immunohistochemical methods and compared with previous data for young adults. For comparison, the biceps brachii muscle of the same subjects was also analysed. The old masseter contained smaller amounts of slow and larger amounts of fast and fetal MyHCs. These differences were region-dependent and were more pronounced in the superficial portion. There was also a larger proportion of “hybrid” fibres, containing two to four MyHC isoforms (42%), compared with the young adult masseter (23%). No such differences were observed between old and young biceps. In contrast to the masseter, the old biceps contained more slow MyHC and less fast MyHC. This investigation demonstrates that the aging process in human skeletal muscle is accompanied by a modification in the muscle phenotype which is both muscle and region specific; a transformation towards a fast and fetal phenotype concomitant with an increased number of fibres with a mixture of different MyHC isoforms in the masseter; and an opposite shift towards a slower phenotype in the biceps brachii. The results might reflect differences between jaw and limb muscles in genetic programs and adaptive responses to changed functional demands following aging.

Fiber Content and Myosin Heavy Chain Composition of Muscle Spindles in Aged Human Biceps Brachii

The present study investigated potential age-related changes in human muscle spindles with respect to the intrafusal fiber-type content and myosin heavy chain (MyHC) composition in biceps brachii muscle. The total number of intrafusal fibers per spindle decreased significantly with aging, due to a significant reduction in the number of nuclear chain fibers. Nuclear chain fibers in old spindles were short and some showed novel expression of MyHC α-cardiac. The expression of MyHC α-cardiac in bag1 and bag2 fibers was greatly decreased in the A region. The expression of slow MyHC was increased in nuclear bag1 fibers and that of fetal MyHC decreased in bag2 fibers whereas the patterns of distribution of the remaining MyHC isoforms were generally not affected by aging. We conclude that aging appears to have an important impact on muscle spindle composition. These changes in muscle spindle phenotype may reflect an age-related deterioration in sensory and motor innervation and are likely to have an impact in motor control in the elderly.

Expression of alpha-cardiac myosin heavy chain in mammalian skeletal muscle

We have investigated the reactivity of different human, rat and cat muscles to a monoclonal antibody directed against human α-cardiac myosin heavy chain. We have found that special fiber subpopulations of human massetr and extraocular muscles, as well as the bag fibers of human, rat and cat muscle spindles, were reactive to this antibody, indicating that these fibers expressed α-cardiac myosin heavy chain or a closely related isoform. This isomyosin was present in the spindle bag fibers at early fetal stages, whereas its expression in masseter and extraocular muscle fibers was not detected during the first 22 weeks of gestation. Our results add to the list of muscle proteins which are expressed in locations or at developmental stages other than those initially described, suggesting that a revision of the present nomenclature of the subgroups of myosin heavy chains might be considered in the future.