Gender- and age-related differences in the regulatory influence of thyroid hormone on the contractility and myosin composition of single rat soleus muscle fibres

 The effects of 4 weeks of thyroid hormone (3,5,3′-triiodothyronine, T₃) treatment on the myosin isoform composition and maximum velocity of unloaded shortening (V ₀) of single soleus muscle fibres of young (3–6 months) and old (20–24 months) female (149 fibres) and male (200 fibres) rats were studied. Gender-related differences in the up-regulation of fast myosin heavy chain (MyHC) and myosin light chain (MyLC) isoforms were observed. In the female hyperthyroid rats, pure type I fibres and fibres co-expressing type I and type IIA MyHC (type I/IIA fibres) predominated. Some fibres expressed an α cardiac-like MyHC isoform either purely (α cardiac-like fibre type) or in co-expression with IIA MyHC (α cardiac-like/IIA fibre type). In the male hyperthyroid rats, on the other hand, all fibres were either type I/IIA or type I/IIAX. The relative quantities of fast MyLC isoforms in type I/IIA and type I/IIAX fibres was higher in female than in male hyperthyroid rats. V ₀ was similar in male and female control rats, and decreased with age in both genders (P<0.001). After T₃ treatment, the average V ₀ increased (P<0.001) in females with a concomitant up-regulation of fast MyHC and fast MyLC isoforms irrespective of age. The average V ₀ of the pooled fibres was higher (P<0.001) in female than in male hyperthyroid rats at both ages. In conclusion, gender- and age-related differences were observed in the regulatory influence of 4 weeks’ T₃ treatment on myosin isoform composition and V ₀ in soleus fibres. These differences are presumably related to an interaction of thyroid and sex hormones in the regulation of myosin gene expression. Received: 10 March 1998 / Received after revision 3 June 1998 / Accepted: 14 August 1998     

Gender-related differences in the regulatory influence of thyroid hormone on the expression of myosin isoforms in young and old rats

The effects of 4 weeks of thyroid hormone (3,5,3′-triiodothyronine, T3) treatment on the expression of myosin heavy chain (MyHC) isoforms were examined in young (3–6 months) and old (20–24 months) female rats, and compared with those in age-matched male rats (Larsson et al. 1995). In control rats, ageing was associated with a type IIA to I MyHC isoform switching in the slow-twitch soleus and a type IIB to IIX MyHC isoform switching in the fast-twitch extensor digitorum longus muscle (EDL). Gender- and muscle-specific differences were observed in the regulation of MyHC isoforms by T3. In the soleus, dramatic downregulation of the type I and upregulation of the type IIA MyHC isoform were observed in both females and males, but upregulation of the IIX MyHC isoform was observed only in male rats. In EDL, T3 treatment had no significant influence on the MyHC isoform composition in the males irrespective of the age of the animal. In the females, on the other hand, T3 treatment resulted in a significant MyHC transformation from IIA to IIB, probably via IIX myosin, in spite of the fact that type IIA mRNA has been reported to be downregulated in both females and males. It is concluded that the regulation of MyHC isoforms by thyroid hormone differs between females and males, presumably as a result of a gender-related difference in the translational or post-translational regulation of MyHC synthesis.     

Thyroid hormone regulation of myosin heavy chain isoform composition in young and old rats,with special reference to IIX myosin

The effects of 4 weeks of thyroid hormone (3,5,3′-triiodothyronine, T₃) treatment on the myosin heavy chain (MHC) composition were compared in the slow-twitch soleus and the fast-twitch extensor digitorum longus (EDL) muscles from young (3–6 months) and old (20–24 months) male albino rats. Four MHC isoforms were separated on silverstained 6% sodium dodecyl sulphate polyacrylamide gel electrophoresis. According to immunoblotting experiments with specific MHC monoclonal antibodies, the four MHCs corresponded to types I, IIB, IIX and IIA. In the soleus, the type I MHC content was higher in the old than in the young animals, and the type IIA content lower. Type IIX myosin was observed in some young control soleus, but not in old ones. After T₃ treatment, the content of type I MHC decreased substantially in both young and old animals and that of type IIA increased. After T₃ treatment, type IIX myosin was observed in both young and old animals, with a slighty higher IIX myosin content in old age, but the age-related different in the contents of types I and IIA was diminished. In EDL, the type IIX MHC content was significantly higher in the old animals, at the significantly by T₃ treatment in EDL, either in young or old animals. In conclusion, an age-related motor unit transformation is observed in both the slow-twich soleus and the fast-twitch EDL and the capacity for MHC isoform switching in response to T₃ treatment is not impaired in old age.     

Effects of removal of weight-bearing function on contractility and myosin isoform composition in single human skeletal muscle cells

The purpose of this study was to investigate the effects of a 6-week period without weight bearing, achieved by bed rest, on the contractile behaviour, myosin isoform expression and myofibrillar protein content of single human muscle fibres. Percutaneous biopsied specimens of the quadriceps muscle were taken from three healthy male volunteers before and at the end of the experimental period. Maximum force normalised to cross-sectional area (specific tension), maximum velocity of unloaded shortening (V  ₀), and myosin heavy chain (MyHC) and light chain (MyLC) isoform composition were measured in single membrane-permeabilised muscle cells obtained from these specimens. At the end of the experimental period, specific tension was reduced (P < 0.001) by 40% and there was a parallel decline in myofibrillar protein content per muscle cell volume. V  ₀ did not change significantly in response to bed rest when data from all muscle cells were pooled. In two of the subjects, however, V  ₀ decreased (P < 0.01–0.001) in muscle cells expressing the β/slow (type I) MyHC isoform, but there was no change in fibres expressing type IIA or a combination of type IIA and IIB MyHCs. The slowing in type I MyHC fibres was associated with a change in the isoform composition of the regulatory MyLC. Received: 5 October 1995/Received after revision and accepted: 3 January 1996     

Cortisone-induced changes in myosin heavy chain distribution in respiratory and hindlimb muscles

In this study the effects of administration of cortisone acetate (100 mg kg^-1 body weight subcutaneously for 11 days) on distribution and cross-sectional area of different fibre types of rat skeletal muscles were investigated. Diaphragm, parasternal intercostal (PI), extensor digitorum longus (EDL) and soleus muscles were examined in cortisone treated animals (CA) in comparison with ad libitum controls (CTRL) and pair-fed (PF) controls. Four fibre types (I or slow and IIA, IIX, IIB or fast) were identified on the basis of their myosin heavy chain composition using a set of monoclonal antibodies. In CA rats the reduction of cross-sectional area was above 30% in IIX fibres of diaphragm, IIB fibres of PI and in all fast fibres of EDL. In all muscles slow fibres were spared from atrophy. Significant variations in fibre type distribution were found in the muscles of CA rats when compared to CTRL. The percentage of IIB fibres decreased in EDL, PI and diaphragm. This decrease was accompanied by an increase in the percentage of IIA fibres in the same muscles. No changes in the percentage of slow fibres and of fast IIX fibres were observed in EDL, PI and diaphragm of CA rats in comparison with CTRL. In soleus of CA rats the proportion of IIA fibres was lower than in CTRL. In EDL of PF rats atrophy of IIA fibres and changes in fibre type distribution were similar to those observed in CA rats. In diaphragm, PI and soleus of PF rats no significant decrease in fibre cross-sectional area nor significant changes in fibre distribution were found in comparison with CTRL rats.     

Fibre type composition of soleus and extensor digitorum longus muscles in normal female inbred Lewis rats

We have analysed the fibre type composition of soleus and extensor digitorum longus (EDL) muscles of normal female 4-6-month-old inbred Lewis rats. This rat strain is used in our ongoing study of the effects of thyroid hormone on myosin heavy chain (MyHC) isoform expression. On the basis of the mATPase reaction, soleus muscles contained 96.1 +/- 2.9% of type 1 fibres supplemented by 2A fibres. EDL muscles contained type 1 (5.5 +/- 1.0%), type 2A (18.8 +/- 1.7%) and type 2B (75.7 +/- 2.2%) fibres. Immunohistochemical analysis and SDS gel electrophoresis confirmed that most fibres in the soleus muscle expressed the type 1 (slow) MyHC isoform and that only a small proportion of fibres expressed the fast 2a MyHC isoform. Immunohistochemical analysis and SDS gel electrophoresis demonstrated that almost half of the 2B fibres of EDL muscles expressed the 2x/d MyHC isoform. In both muscle types, many fibres expressed more than one MyHC isoform. The content of slow fibres in the soleus muscle of female inbred Lewis rats was slightly higher than that reported for Wistar rats, but was considerably higher than that of Sprague-Dawley rats, whereas substantial differences were not found in the proportion of slow and fast fibre types in EDL muscles in these strains.     

Reinnervation of denervated extensor digitorum longus of the rat by the nerve of the soleus does not induce the type I myosin synthesis directly but through a sequential transition of type II myosin isoforms.

The fast-contracting extensor digitorum longus (EDL) muscle of 1-month-old rats was denervated and reinnervated by the nerve innervating the slow-contracting soleus muscle. After variable periods of time, the myosin isoform content of the EDL was analyzed by sensitive electrophoretic techniques, which allowed to discriminate between the slow-type I and the three, IIA, (IID or IIX) and IIB, fast-type II myosin isoforms. Compared to the control EDL, which contains predominantly the IIB isoform, the operated muscles contained variable proportions of all the isoforms. Analysis of the results leads us to conclude that reinnervation of EDL induces a sequential transition of myosin isoforms: IIB----(IID or IIX)----IIA----I.     

Fibre size and metabolic properties of myosin heavy chain-based fibre types in rat skeletal muscle

Cross-sectional area (CSA), succinate dehydrogenase (SDH), and -glycerophosphate dehydrogenase (GPD) activities were measured in single fibres of adult rat medial gastrocnemius muscle (deep region) using quantitative histochemical procedures. The same fibres were identified in serial sections stained by immunohistochemistry with monoclonal antibodies specific for selected myosin heavy chain (MyHC) isoforms. The values of CSA, SDH and GPD activities formed a continuum, but significant differences in mean values were observed among fibre types of presumed homogeneous MyHC content. Type IIA fibres were the smallest, type IIB fibres were the largest, and type I and IIX fibres were intermediate. Type IIA fibres had the highest SDH activity, followed in rank order by type IIX, type I and type IIB. The average GPD activity was ranked according to fibre type such that IIB>IIX>IIA>I. Hybrid fibres co-expressing two MyHC isoforms generally showed intermediate mean CSA, SDH a nd GPD values lying between their respective pure MyHC fibre types. Across all fibres, there was an inverse relationship between SDH activity and CSA and between GPD and SDH activities, and a positive correlation between GPD and CSA. Moreover, a significant interdependence between CSA, SDH activity, GPD activity and MyHC content existed on a fibre-to-fibre basis, suggesting that the MyHC isoform expressed in a fibre is associated with differences in size, oxidative and glycolytic capabilities of muscle fibres. In fact, most of the fibres could be discriminated into discrete groups with the same MyHC content when their CSA, SDH and GPD values were considered together.     

Adaptation of rat extensor digitorum longus muscle to gamma irradiation and overload

The right extensor digitorum longus (EDL) muscle of growing male rats was overloaded by ablation of its synergist tibialis anterior (TA) muscle. Four weeks later, the overloaded muscle was heavier and contained larger type IIA, IIX and IIB fibres than either untreated contralateral muscle or control muscle from an untreated animal. The myonuclear-to-myoplasmic volume ratio was maintained in the overloaded muscle. Overloaded EDL muscle, previously subjected to a dose of irradiation sufficient to sterilise satellite cells, and EDL muscle which had been only irradiated, were significantly lighter and contained significantly smaller fibres than controls, though a significant amount of normal EDL muscle growth did occur following either treatment. The myonuclear-to-myoplasmic volume ratio of the irradiated muscles was smaller than in controls. Overloaded muscle, with or without prior irradiation, possessed a smaller proportion of fibres containing IIB myosin heavy chain (MHC) and a larger proportion of fibres containing IIA and IIX MHC; a significant percentage of these fibres coexpressed either type IIA and IIX MHC or type IIX and IIB MHC. Thus in the absence of satellite cell mitosis, muscles of young rats possess a limited capacity for normal growth but not for compensatory hypertrophy. Adaptations in MHC gene expression to chronic overload are completely independent of satellite cell activity.     

Contractility and myosin isoform compositions of skeletal muscles and muscle cells from rats treated with thyroid hormone for 0, 4 and 8 weeks

The effects of 4 and 8 weeks of thyroidhormone (3,5,3-triiodothyronine, T3) treatment on skeletalmuscles of young (3--6 months) male Wistar rats were investigatedin the present study. In the slow-twitch soleus, contraction andhalf-relaxation times of the isometric twitch were significantlyshorter in hyperthyroid rats than in the control group, andtwitch duration was shorter in rats treated with T3 for 8 weeksthan for 4 weeks. All single soleus muscle fibres fromhyperthyroid rats co-expressed types I and IIA myosin heavychains (type I/IIA fibres) or type I, IIA and IIX myosin heavychains (type I/IIAX fibres), while only type I MyHC fibres wereisolated from the controls. A significantly higher content oftype IIA myosin heavy chain and fast myosin light chain isoformswas observed in soleus fibres from the 8-week than from 4-week T3group. There was no significant difference in maximum velocity ofunloaded shortening (V0) between type I myosin heavy chain fibresfrom controls (1.12 ± 0.46 muscle lengths s–1, n = 48)and type I/IIA myosin heavy chain fibres from the 4 – (1.09± 0.36 muscle lengths s–1, n = 33) and 8-week (1.03± 0.31 muscle lengths s–, n = 31) groups, but typeI/IIAX fibres from the 8-week T3 group had significantly higherV0 (1.56 ± 0.10, n = 5) than type I from control and typeI/IIA from hyperthyroid rats. In the fast-twitch extensordigitorum longus, neither myosin isoform composition, twitchduration nor V0 was affected by 4 or 8 weeks of T3 exposure. Inconclusion, a dramatic and exposure duration-dependent change inthe contractile speed of the isometric twitch and the expressionof fast myosin isoforms was observed in soleus, but not inextensor digitorum longus, in response to T3 treatment. Long-termT3 treatment had relatively less influence, however, on V0 at thesingle cell level in spite of the dramatic increase in fastmyosin isoforms     

Laryngeal muscle fibre types

The internal laryngeal muscles have evolved to subserve the highly specialized functions of airways protection, respiration, and phonation. Their contractile properties, histochemistry, biochemical properties, myosin heavy chain (MyHC) expression and their regulation by nerves and hormones are reviewed and compared with limb muscle fibres. Cricothyroid, the vocal cord tensor, is limb-like in MyHC composition and fibre type properties, while the vocal fold abductor and adductors are allotypically different, with capacity for expressing an isoform of MyHC that is kinetically faster than the fastest limb MyHC. In rats and rabbits the faster isoform is the extraocular (EO) MyHC, while in carnivores, it is the IIB MyHC. These adaptations enable the abductor and adductor muscles to remain always faster than the cricothyroid as the latter changes in speed during evolution to match changing metabolic and respiratory rates in relation to scaling with body mass. Such phylogenetic plasticity is vital to the airways protection and respiratory functions of these muscles. The posterior cricoarythenoid, the abductor muscle, is tonically driven during expiration, and consequently has a slower fibre type profile than the principal adductor, the thyroarythenoid. The human thyroarythenoid appears not to express EO or IIB MyHC significantly, but is unique in expressing the slow-tonic MyHC. The concepts of allotype and phylogenetic plasticity help to explain differences in fibre type between limb and laryngeal muscles and between homologous laryngeal muscles in different species. Laryngeal muscle fibres exhibit physiological plasticity as do limb muscles, being subject to neural and hormonal modulation.     

Laryngeal muscle fibre types

The internal laryngeal muscles have evolved to subserve the highly specialized functions of airways protection, respiration, and phonation. Their contractile properties, histochemistry, biochemical properties, myosin heavy chain (MyHC) expression and their regulation by nerves and hormones are reviewed and compared with limb muscle fibres. Cricothyroid, the vocal cord tensor, is limb‐like in MyHC composition and fibre type properties, while the vocal fold abductor and adductors are allotypically different, with capacity for expressing an isoform of MyHC that is kinetically faster than the fastest limb MyHC. In rats and rabbits the faster isoform is the extraocular (EO) MyHC, while in carnivores, it is the IIB MyHC. These adaptations enable the abductor and adductor muscles to remain always faster than the cricothyroid as the latter changes in speed during evolution to match changing metabolic and respiratory rates in relation to scaling with body mass. Such phylogenetic plasticity is vital to the airways protection and respiratory functions of these muscles. The posterior cricoarythenoid, the abductor muscle, is tonically driven during expiration, and consequently has a slower fibre type profile than the principal adductor, the thyroarythenoid. The human thyroarythenoid appears not to express EO or IIB MyHC significantly, but is unique in expressing the slow‐tonic MyHC. The concepts of allotype and phylogenetic plasticity help to explain differences in fibre type between limb and laryngeal muscles and between homologous laryngeal muscles in different species. Laryngeal muscle fibres exhibit physiological plasticity as do limb muscles, being subject to neural and hormonal modulation.     

Editor's Choice

Aim: To investigate the hypothesis that mitochondrial efficiency (i.e. P/O ratio) is higher in type I than in type II fibres during submaximal rates of respiration. Methods: Mitochondria were isolated from rat soleus and extensor digitorum longus (EDL) muscles, representing type I and type II fibres, respectively. Mitochondrial efficiency (P/O ratio) was determined with pyruvate (Pyr) or palmitoyl‐l ‐carnitine (PC) during submaximal (constant rate of adenosine diphosphate infusion) and maximal (V_max, state 3) rates of respiration and fitted to monoexponential functions. Results: There was no difference in V_max between PC and Pyr in soleus but in EDL V_max with PC was only 58% of that with Pyr. The activity of 3‐hydroxyacyl‐CoA dehydrogenase was threefold higher in soleus than in EDL. P/O ratio at V_max was 8–9% lower with PC [2.33 ± 0.02 (soleus) and 2.30 ± 0.02 (EDL)] than with Pyr [2.52 ± 0.03 (soleus) and 2.54 ± 0.03 (EDL)] but not different between the two muscles (P > 0.05). P/O ratio was low at low rates of respiration and increased exponentially when the rate of respiration increased. The asymptotes of the curves were similar to P/O ratio at V_max. P/O ratio at submaximal respirations was not different between soleus and EDL neither with Pyr nor with PC. Conclusion: Mitochondrial efficiency, as determined in vitro, was not significantly different in the two fibre types neither at V_max nor at submaximal rates of respiration. The low V_max for PC oxidation in EDL may relate to low activity of β‐oxidation.     

Relationship between skin blood flow and sweating rate in prepubertal boys and young men

We have examined the effect of male sexual hormones on the regeneration of skeletal muscles. Degeneration/regeneration of the left soleus and extensor digitorum longus muscles (EDL) of Wistar male rats was induced by an injection of snake venom (2 microg, Notechis scutatus scutatus). During the muscle regeneration (25 days), rats were treated with either oil (CON), nandrolone (NAN), NAN combined with exercise (NAN + EXE) or were castrated (CAS). Muscle growth and myosin heavy chain (MyHC) isoform content of regenerating muscles were studied. Castration altered the concentrations of MyHC in venom-treated EDL (P < 0.01) and soleus (P < 0.05). NAN increased the mass (P < 0.01) of regenerating soleus and decreased the relative amount of fast MyHC protein (% of total, P < 0.05). The effect of NAN + EXE on the fast MyHC proteins of venom-treated soleus was opposite (P < 0.05). NAN and NAN + EXE were without effect on the regenerating EDL (P > 0.05). In conclusion, it is possible that male sexual hormones play a role in the growth (synthesis of contractile proteins) of regenerating muscles in rat. In addition, contrary to NAN + EXE, NAN could be beneficial to soleus regeneration.     

Differences in ultrastructural and metabolic profiles within the same type of fibres in various muscles of young and adult rats

Single fibres from tibialis anterior, extensor digitorum longus, gastrocnemius and soleus muscles in young (4–week–old) and adult (35–week–old) Wistar male rats were classified into three types on the basis of their enzyme–histochemical features: slow–twitch oxidative (SO), fast–twitch oxidative and glycolytic (FOG) and fast–twitch glycolytic (FG) fibres. Ultrastructural (volume density of mitochondria: V_mt and Z line width) and metabolic (phosphofructokinase: PFK and succinate dehydrogenase: SDH activities) profiles were measured. PFK activity in all types of fibres¹ was higher in adult rats, and the difference between the two age–groups (adult/young) was largest between FG, FOG and SO fibres respectively. SDH activity and V_mt were lower in adult rats in a similar way in all fibres. A significant positive correlation was observed between the V_mt and SDH activity in both age–groups. This positive correlation was very specific in fast–twitch and slow–twitch fibres. Changes in the V_mt did not relate directly to the changes in fibre cross–sectional area. The overall pattern indicates that glycolytic capacity of fast–twitch fibres in flexor muscles (TA and EDL) is higher than in extensor muscles (GC and SOL), and that oxidative capacity of all types of fibre in extensor muscles is higher than in flexor muscles. These profiles were changed by growth, and may be related to the specific differences in pattern of activity of each skeletal muscle, and may reflect differences in the recruitment order of different muscles.     

Muscle fibre number is a possible determinant of muscle fibre composition in rats

The purpose of the present study was to investigate whether the muscle fibre composition is related to the number of muscle fibres. To resolve this issue, we developed fast-twitch fibre dominant rats (FFDR) by selective breeding and compared the findings to those of control rats (CR) obtained by random breeding. Percentage of type I fibres of the deep portion of gastrocnemius (DG), soleus (SOL), vastus intermedius (VI), adductor longus (AL), and biceps brachii (BB) muscles in FFDR were lower than CR. Percentage of type IIB fibres in DG, VI and AL and percentage of type IIA fibres of SOL in FFDR were higher than CR. However, fibre composition of plantaris (PLAN), extensor digitorum longus (EDL), rectus abdominis (RA), diaphragm (DIA), and palmaris longus (PL) muscles in FFDR were identical with CR. Total fibre numbers on the cross-sectional area in SOL, PLAN, EDL, AL and PL were counted. Numbers of type I fibres of all those muscles in FFDR were not different from CR. Numbers of type IIA fibres of SOL and AL and of type IIB fibres of AL in FFDR were greater than CR, but there were no significant differences in the number of type IIA or type IIB fibres of PLAN, EDL or PL between the two groups. Based on these observations, it is suggested that there are pleiotropic and muscle-specific effects on muscle fibre composition. In addition, the number of type II fibres is a possible determinant of muscle fibre composition.     

'Fast' and 'slow' muscle fibres in hindlimb muscles of adult rats regenerate from intrinsically different satellite cells

Myosin heavy chain (MyHC) expression was examined in regenerating fast extensor digitorum longus (EDL) and slow soleus (SOL) muscles of adult rats. Myotoxic bupivacaine was injected into SOL and EDL and the muscles were either denervated or neuromuscularly blocked by tetrodotoxin (TTX) on the sciatic nerve. Three to 10 or 30 days later, denervated SOL or EDL, or innervated but neuromuscularly blocked EDL received a slow 20 Hz stimulus pattern through electrodes implanted on the muscles or along the fibular nerve to EDL below the TTX block. In addition, denervated SOL and EDL received a fast 100 Hz stimulus pattern. Denervated EDL and SOL stimulated with the same slow stimulus pattern expressed different amounts of type 1 MyHC protein (8% versus 35% at 10 days, 13% versus 87% at 30 days). Stimulated denervated and stimulated innervated (TTX blocked) EDL expressed the same amounts of type 1, 2A, 2X and 2B MyHC proteins. Cross-sections treated for in situ hybridization and immunocytochemistry showed expression of type 1 MyHC in all SOL fibres but only in some scattered single or smaller groups of fibres in EDL. The results suggest that muscle fibres regenerate from intrinsically different satellite cells in EDL and SOL and within EDL. However, induction by different extrinsic factors arising in extracellular matrix or from muscle position and usage in the limb has not been excluded. No evidence for nerve-derived trophic influences was obtained.     

Differential effects of a six-day immobilization on newborn rat soleus muscles at two developmental stages

Our objective was to determine the effects of a six-day immobilization on the musculoskeletal system of the rat during postnatal development at two key periods when the states of innervation are known to be different. This work was undertaken on the soleus muscle since it is well known that postural slow muscles show marked changes after a period of disuse. Thus, the soleus muscle was immobilized in a shortened position either when the innervation was polyneuronal or monosynaptic, respectively from 6 to 12 and from 17 to 23 days. The muscle modifications were followed by ATPase staining and myosin heavy chain (MyHC) isoform identification using monoclonal antibodies and SDS–PAGE. The functional properties of skinned fibre bundles were established by calcium/strontium (Ca/Sr) activation characteristics. In control muscles the maturation was characterized by a progressive increase of adult MyHCs (I and IIA) concomitant with a decrease in both the MyHC neo and the Ca affinit y. Between 6 to 12 days, immobilization of the limb induced an increase in histochemical type IIC fibres. Using antibodies we identified new fibre types, classified as a function of their MyHC isoform co-expression. We observed an increase in expression of both MyHC neo and Ca affinity. From 17 to 23 days, the immobilization induced an increase in Ca affinity and marked changes in the MyHC isoform composition: disappearance of MyHC neo and expression of the fast MyHC IIB isoform, which in normal conditions is never expressed in the soleus muscle. We conclude that an immobilization imposed during polyneuronal innervation delays the postnatal maturation of the soleus muscle, whereas when the immobilization is performed under monosynaptic innervation the muscle evolves towards a fast phenotype using a default pathway for MyHC expression.