Fiber Myosin Heavy Chain Research Articles

Unloaded shortening velocity and myosin heavy chain variations in human laryngeal muscle fibers.

Myosin description in human laryngeal muscles is incomplete, but evidence suggests the presence of type I, IIA, IIX, and tonic myosin heavy chain (MHC) fibers. This study describes the unloaded shortening velocity (V0) of chemically skinned laryngeal muscle fibers measured by the slack test method in relation to MHC content. Skeletal fibers from human laryngeal and limb muscle biopsy specimens were obtained for determination of V0, and subsequently, glycerol-sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the MHC isoform content. The fibers from human limb muscle had shortening speeds similar to those in previous reports on human skeletal fibers. Type I, IIA, and IIX fibers of laryngeal muscle had shortening speeds similar to those of fibers from limb muscle, but laryngeal fibers with heterogeneous MHC expression had a wide range of shortening speeds, some being nearly twice as fast as limb fibers. In addition, MHC isoform bands from human extraocular muscle comigrated with some bands from laryngeal muscle--a finding suggesting that extraocular myosin may also be expressed.

Skeletal muscle adaptations following spinal cord contusion injury in rat and the relationship to locomotor function: a time course study.

Experimental spinal cord injury (SCI) via contusion of moderate severity results in residual locomotor deficits, including a lack of coordination and trunk stability. Given that muscle contractile properties and fiber composition adapt to reduced neural input and/or weight bearing, contusion-induced locomotor deficits may reflect changes in hindlimb skeletal muscle. Therefore, we examined muscle adaptations during early (1 week), intermediate (3 week), and late (10 week) stages of motor recovery after moderate SCI. Forty-two Sprague Dawley rats underwent SCI via 1.1mm cord displacement with the OSU impact device or served as age and weight-matched or laminectomy controls. Subsets of rats had soleus (SOL) in vitro physiological testing or SOL and extensor digitorum longus (EDL) myosin heavy chain (MHC) fiber type analysis. At 1 week post-SCI during paralysis/paresis, a significant decrease in wet weight occurred in the plantaris, medial/lateral gastrocnemius (MG/LG), tibialis anterior, and SOL. Changes in contractile properties of the SOL did not accompany muscle wet weight changes. By 3 weeks, the loss of weight-bearing activity early after SCI induced significant decreases in SOL peak twitch and peak tetanic tension as well as significantly greater IIx MHC expression in the EDL. By 10 weeks post-SCI, after several weeks of weight supported stepping, muscle wet weight, contractile properties and MHC composition returned to baseline levels except for MG/LG atrophy. Thus, muscle plasticity appears to be extremely sensitive to locomotor deficits and their resolution after moderate spinal cord contusion.

Functional properties of slow and fast gastrocnemius muscle fibers after a 17-day spaceflight

The purpose of this investigation was to study the effects of a 17-day spaceflight on the contractile properties of individual fast- and slow-twitch fibers isolated from biopsies of the fast-twitch gastrocnemius muscle of four male astronauts. Single chemically skinned fibers were studied during maximal Ca2+-activated contractions with fiber myosin heavy chain (MHC) isoform expression subsequently determined by SDS gel electrophoresis. Spaceflight had no significant effect on the mean diameter or specific force of single fibers expressing type I, IIa, or IIa/IIx MHC, although a small reduction in average absolute force (P(o)) was observed for the type I fibers (0.68 +/- 0.02 vs. 0.64 +/- 0.02 mN, P < 0.05). Subject-by-flight interactions indicated significant intersubject variation in response to the flight, as postflight fiber diameter and P(o) where significantly reduced for the type I and IIa fibers obtained from one astronaut and for the type IIa fibers from another astronaut. Average unloaded shortening velocity [V(o), in fiber lengths (FL)/s] was greater after the flight for both type I (0.60 +/- 0.03 vs. 0.76 +/- 0.02 FL/s) and IIa fibers (2.33 +/- 0.25 vs. 3.10 +/- 0.16 FL/s). Postflight peak power of the type I and IIa fibers was significantly reduced only for the astronaut experiencing the greatest fiber atrophy and loss of P(o). These results demonstrate that 1) slow and fast gastrocnemius fibers show little atrophy and loss of P(o) but increased V(o) after a typical 17-day spaceflight, 2) there is, however, considerable intersubject variation in these responses, possibly due to intersubject differences in in-flight physical activity, and 3) in these four astronauts, fiber atrophy and reductions in P(o) were less for slow and fast fibers obtained from the phasic fast-twitch gastrocnemius muscle compared with slow and fast fibers obtained from the slow antigravity soleus [J. J. Widrick, S. K. Knuth, K. M. Norenberg, J. G. Romatowski, J. L. W. Bain, D. A. Riley, M. Karhanek, S. W. Trappe, T. A. Trappe, D. L. Costill, and R. H. Fitts. J Physiol (Lond) 516: 915-930, 1999].

Intermuscular and intramuscular differences in myosin heavy chain composition of the human masticatory muscles

Among and within the human masticatory muscles a large number of anatomical differences exists indicating that different muscles and muscle portions are specialized for certain functions. In the present study we investigated whether such a specialization is also reflected by intermuscular and intramuscular differences in fibre type composition and fibre cross-sectional area. Fibre type compositions and fibre cross-sectional areas of masticatory muscles were determined in eight cadavers using monoclonal antibodies against myosin heavy chain (MyHC). The temporalis, masseter and pterygoid muscles could be characterized by a relatively large number of fibres containing more than one MyHC isoform (hybrid fibres). In these muscles a large number of fibres expressed MyHC-I, MyHC-fetal and MyHC-cardiac α. Furthermore, in these muscles type I fibres had larger cross-sectional areas than type II fibres. In contrast, the mylohyoid, geniohyoid and digastric muscle were characterized by less hybrid fibres, and by less fibres expressing MyHC-I, MyHC-fetal, and MyHC-cardiac α, and by more fibres expressing MyHC-IIA; the cross-sectional areas of type I and type II fibres in these muscles did not differ significantly. Compared to the masseter and pterygoid muscles, the temporalis had significantly larger fibres and a notably different fibre type composition. The mylohyoid, geniohyoid, and digastric muscles did not differ significantly in their MyHC composition and fibre cross-sectional areas. Also intramuscular differences in fibre type composition were present, i.e., a regionally higher proportion of MyHC type I fibres was found in the anterior temporalis, the deep masseter, and the anterior medial pterygoid muscle portions; furthermore, significant differences were found between the bellies of the digastric.

Sexually dimorphic expression of myosin heavy chains in the adult mouse masseter

Little is known regarding the role of androgenic hormones in the maintenance of myosin heavy chain (MHC) composition of rodent masticatory muscles. Because the masseter is the principal jaw closer in rodents, we felt it was important to characterize the influence of androgenic hormones on the MHC composition of the masseter. To determine the extent of sexual dimorphism in the phenotype of masseter muscle fibers of adult (10-mo-old) C57 mice, we stained tissue sections with antibodies specific to type IIa and IIb MHC isoforms. Females contain twice as many fibers containing the IIa MHC as males, and males contain twice as many fibers containing the IIb MHC as females. There is a modest amount of regionalization of MHC phenotypes in the mouse masseter. The rostral portions of the masseter are composed mostly of type IIa fibers, whereas the midsuperficial and caudal regions contain mostly type IIb fibers. Using immunoblots, we showed that castration results in an increase in the expression of type IIa MHC fibers in males. Ovariectomy has no effect on the fiber type composition in females. We conclude that testosterone plays a role in the maintenance of MHC expression in the adult male mouse masseter.

Persistence of myosin heavy chain-based fiber types in innervated but silenced rat fast muscle

Myosin heavy chain (MHC) profile and size of fibers in deep and superficial regions of the adult rat medial gastrocnemius (MG) were determined after 4, 15, 30, and 60 days of inactivity induced by spinal cord isolation (SI). After 4 days, fiber size decreased by 33 to 50% and 36 to 46% in deep and superficial regions, whereas MHC composition was unaffected. By 15 days, these values were 45 to 78% and 51 to 69%, and MHC composition was shifting toward faster isoforms. By 60 days, there were no pure type I MHC fibers and increases from 1 to 18% and 78 to 93% in pure type IIb fibers in deep and superficial regions. The percentage of type I MHC (gel electrophoresis) was∼10 and ∼3%, and of type IIb ∼40 and ∼60% in control and 60-day SI rats. Thus, adaptations in the MHC molecule occurred at a slower rate and for a longer duration than the atrophic response. © 2000 John Wiley & Sons, Inc. Muscle Nerve 23: 735–747, 2000

Persistence of myosin heavy chain-based fiber types in innervated but silenced rat fast muscle.

Myosin heavy chain (MHC) profile and size of fibers in deep and superficial regions of the adult rat medial gastrocnemius (MG) were determined after 4, 15, 30, and 60 days of inactivity induced by spinal cord isolation (SI). After 4 days, fiber size decreased by 33 to 50% and 36 to 46% in deep and superficial regions, whereas MHC composition was unaffected. By 15 days, these values were 45 to 78% and 51 to 69%, and MHC composition was shifting toward faster isoforms. By 60 days, there were no pure type I MHC fibers and increases from 1 to 18% and 78 to 93% in pure type IIb fibers in deep and superficial regions. The percentage of type I MHC (gel electrophoresis) was approximately 10 and approximately 3%, and of type IIb approximately 40 and approximately 60% in control and 60-day SI rats. Thus, adaptations in the MHC molecule occurred at a slower rate and for a longer duration than the atrophic response.

Fibre types and myosin heavy chain expression in the ocular medial rectus muscle of the adult rat.

Myosin heavy chain (MHC) expression was determined immunohistochemically in individual muscle fibre types characterised by activities of ATPase and the key oxidative and glycolytic enzymes in rat ocular medial rectus (MR) muscles. In the global layer (GL), glycolytic activity of muscle fibres was higher and oxidative activity lower, than in the orbital layer (OL). Muscle fibres in the former displayed rosette-like organisation with a slow fibre surrounded by several fast fibres, which expressed either MHCIIa or MHCIIb, but many co-expressed both isoforms. In the OL some slow fibres co-expressed MHCIIa. Extraocular MHC isoform (MHCeom) could not be determined immunohistochemically and no pure MHCIIx/d containing fibres were found, suggesting that these isoforms, demonstrated electrophoretically, are co-expressed with others. Slow muscle fibres in both layers co-expressed MHCbeta slow, MHCalpha cardiac and MHC-slow tonic. Neonatal isoform (MHCneo) was co-expressed in several fast and slow muscle fibres in the orbital, but not global layer. Slow fibres in the GL displayed very low oxidative activity. Electrophoretic analysis of ocular MR muscle homogenates revealed that about 50% of total MHC was MHCIIb, MHCeom was quite prominent (25%), and MHCIIa, MHCIIx/d and MHCI contributed each about 8%. MHCneo, MHCslow tonic and MHCalpha cardiac could not be identified as separate bands.

Standardisation of muscular biopsy of gluteus medius in French trotters.

Morphometric measurements were taken from 41 French trotters of various ages and both sexes. Biopsy location was determined for the dorsal compartment as being one-third of the distance from the tuber sacrale to the tuber coxae and for the ventral compartment as being one-third of the distance from the tuber coxae to the caudal Cd1-Cd2 intervertebral joint. Ten horses were biopsied at these 2 sites at a sampling depth equal to half the total depth of the compartment as measured by ultrasonography. The percentage of slow and fast myosin heavy chain fibres was measured by using an immunoenzymatic method. The depth of the dorsal and ventral compartments of the gluteus medius was significantly greater in males than in females. The depth of the ventral compartment was greater in the case of a straight hip than of a wide hip and was greater in young horses than in older horses. The ventral and the dorsal compartments were composed in the mid-portion of 81.3 and 75.6% of fast myosin heavy chains, respectively. It was concluded that the locations and sampling depths in the gluteus medius could be standardised in French trotters by taking into account anatomical references, sex, age and width of the hip.

Contractile properties and protein isoforms of single skeletal muscle fibers from 12- and 30-month-old Fischer 344 Brown Norway F1 hybrid rats

This study characterizes single skeletal muscle fiber contractile properties and myosin heavy chain (MHC) isoform compositions from the soleus (SOL) and deep portion of the lateral head of the gastrocnemius (RG) muscles of 12- and 30-month-old Fischer 344 Brown Norway F1 hybrid rats (FBN F1). Thirty months of age is approximately the age of 50% survival for the FBN F1 rat. For type I MHC individual fibers from the SOL of 30-month-old animals, the diameter was 88 +/- 2 microns, peak active force was 4.4 +/- 0.2 x 10(-4) N, peak specific tension (P0) was 76 +/- 5 kN/m2, and maximal unloaded shortening velocity (V0) was 0.98 +/- 0.09 fl/s. The type I MHC fibers from the SOL of 12-month-old animals had similar properties with the exception of P0 which was 92 +/- 4 kN/m2 and V0 which was 1.65 +/- 0.12 fl/s. Contractile properties of the RG MHC type I fibers were not significantly different from MHC type I fibers from the SOL in both age groups. The V0 of the RG type IIa MHC fibers from the 12-month animals (4.05 +/- 0.36 fl/s) and 30-month animals (3.55 +/- 0.41 fl/s) and of fibers co-expressing type I MHC and type IIa MHC from 12-month animals (4.21 +/- 0.55 fl/s) and 30-month animals (2.22 +/- 0.27 fl/s) were significantly faster than that of MHC type I fibers of the respective age group. In conclusion, skeletal fibers from the 12- and 30-month-old FBN F1 rats demonstrate fiber-type specific properties with a close relationship between the MHC isoform composition and the V0.

Distribution of fast myosin heavy chain-based muscle fibres in the gluteus medius of untrained horses: mismatch between antigenic and ATPase determinants.

The distribution of muscle fibres classified on the basis of their content of different myosin heavy chain (MHC) isoforms was analysed in muscle biopsies from the gluteus medius of adult untrained horses by correlating immunohistochemistry with specific anti-MHC monoclonal antibodies and standard myofibrillar ATPase (mATPase) histochemistry. Percutaneous needle biopsies were taken at 3 depths (20, 40 and 60 mm) from 4 4-y-old Andalusian stallions. The percentage of 'pure' I MHC fibres increased whereas that for pure IIX MHC fibres decreased from the most superficial to the deepest sampling site. Within the fast fibres, types IIA and IIAX MHC-classified fibres were proportionately more abundant in the deepest sampling site than in the superficial region of the muscle. The immunohistochemical and histochemical characterisation of a large number of single fibres (n = 1375) was compared and correlated on a fibre-to-fibre basis. The results showed that 40% of the fibres analysed were pure type I (expressing only MHC-I); they showed correct matching between their antigenic and mATPase determinants. In contrast, within the fast fibres, a considerable proportion of fibres were found showing a mismatch between their immunohistochemical and mATPase profiles. The most common mismatched fibre phenotypes comprised fibres displaying coexpression of both fast MHCs when analysed by immunocytochemistry, but showing an mATPase profile similar to typical IIX fibres (moderate mATPase reaction after preincubation at pH 4.4). Considered altogether, the total mismatched fibres represented only 4.2% of the whole fast fibre population in the superficial region of the muscle, but their proportion increased to 15.6% and 38.4% in the middle and deep regions, respectively, of gluteus medius. It is concluded that a considerable number of hybrid fast MHC IIAX fibres are present in the gluteus medius of untrained horses, suggesting that equine type II fibres have probably been misclassified in numerous previous publications based on the use of histochemistry alone. This has important implications in attempts to study the physiological properties of fast fibre types adequately in horses.

Doxorubicin chemomyectomy as a treatment for cervical dystonia: histological assessment after direct injection into the sternocleidomastoid muscle.

The sternocleidomastoid muscle (SCM) is one of the major muscles involved in producing abnormal head position in cervical dystonia patients. This study tested whether doxorubicin chemomyectomy, direct injection of doxorubicin into the SCM to permanently remove muscle fibers, has the potential to be a nonsurgical, permanent treatment for cervical dystonia. The right SCM of rabbits was injected with either 1 or 2 mg doxorubicin. Animals were sacrificed 1-2 months postinjection. The SCM was prepared for histological examination of muscle fiber loss and fiber type composition. In all cases, direct injection of doxorubicin resulted in significant decreases in total muscle cross-sectional areas ranging from 75% up to 98%. Individual myofiber cross-sectional areas were smaller than normal after 2 mg doxorubicin treatment, but similar to normal fiber size after 1 mg doxorubicin. There were increased numbers of myofibers that expressed slow and neonatal myosin heavy chain isoforms in these remaining muscle fibers compared to the untreated SCM on the contralateral side. Developmental myosin heavy chain (MHC) was also present in 53% of the remaining myofibers of the treated muscles. The fiber type composition of muscles contralateral to the doxorubicin injections was compared to the fiber type composition of SCM from normal, untreated controls; no difference was seen in the proportions of fast, slow, and neonatal MHC fiber types in these SCM muscles. In summary, the direct injection of doxorubicin into the SCM resulted in significant muscle loss. This supports the use of doxorubicin chemomyectomy as a potential permanent, nonsurgical treatment for cervical dystonia.

Myosin heavy chain composition in rat laryngeal muscles after denervation.

The effects of denervation on myosin heavy chain (MHC) expression in specific laryngeal muscles are characterized using gel electrophoresis. Observed temporal changes in MHC composition will then be used as a biologic marker in studies designed to develop strategies for laryngeal reinnervation and gene therapy. Animal study using an adult rat model for laryngeal paralysis. In anesthetized rats the left recurrent and superior laryngeal nerve were divided. Animals were survived for 7, 14, 28, 90, and 180 days. Animals were euthanized and the thyroarytenoid (TA), vocalis (VOC), posterior cricoarytenoid (PCA), lateral cricoarytenoid (LCA), and cricothyroid (CT) muscle excised. Each muscle was processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and densitometric measurements were obtained to determine composition of MHC fiber types. The changes in relative MHC composition are described for each specific laryngeal muscle. In general, a decrease in type IIB and an increase in IIA and IIX are seen after denervation. Expression of IIL in the denervated condition is variable and the relative change in type I is minimal. This study supports previous work using rat soleus muscle in which IIA/IIX expression is favored in conditions with decreased neuromuscular activity, and conversely, IIB expression is activity dependent. Expression of type I appears to be independent of neural activity. Further study will be undertaken to quantify expression of MHC components and to study factors modulating expression.

Effects of age and gender on shortening velocity and myosin isoforms in single rat muscle fibres.

The maximum velocity of unloaded shortening (V0) and the myosin heavy chain (MyHC) and light chain (MyLC) isoform composition were determined in single fibres from soleus and extensor digitorum longus (EDL) muscles of male and female rate 3-6 and 22-24 months old. In the soleus muscle, the beta/slow (type I MyHC) isoform predominated in both young and old animals, irrespective of gender. In the EDL, fibres expressing type IIX MyHC or a combination of IIX and IIB (IIXB) MyHC isoforms were predominant in old rats, while type IIB MyHC fibres predominated in young individuals of both genders. The V0 of soleus fibres expressing the type I MyHC isoform decreased (P < 0.001) by 40% with age in spite of an unchanged MyLC composition. In the EDL, the V0 of fibres expressing IIX, IIXB and IIB MyHC isoforms did not changes with age or differ between males and females. In conclusion, similar age-related changes in V0 and MyHC composition were observed in single muscle cells from both male and female rats. The present results demonstrate that the relationship between V0 and MyHC isoform composition at the single fibre level is similar in male and female rats, and that similar qualitative changes take place during ageing in both genders.

Contractile function of single muscle fibers after hindlimb unweighting in aged rats.

This investigation determined how muscle atrophy produced by hindlimb unweighting (HU) alters the contractile function of single muscle fibers from older animals (30 mo). After 1 wk of HU, small bundles of fibers were isolated from the soleus muscles and the deep region of the lateral head of the gastrocnemius muscles. Single glycerinated fibers were suspended between a motor lever and force transducer, functional properties were studied, and the myosin heavy chain (MHC) composition was determined electrophoretically. After HU, the diameter of type I MHC fibers of the soleus declined (88 +/- 2 vs. 80 +/- 4 microns) and reductions were observed in peak active force (47 +/- 3 vs. 28 +/- 3 mg) and peak specific tension (Po; 80 +/- 5 vs. 56 +/- 5 kN/m2). The maximal unloaded shortening velocity increased. The type I MHC fibers from the gastrocnemius showed reductions in diameter (14%), peak active force (41%), and Po (24%), whereas the type IIa MHC fibers showed reductions in peak active force and Po. Thus 1 wk of inactivity has a significant effect on the force-generating capacity of single skeletal muscle fibers from older animals in a fiber type-specific manner (type I MHC > type IIa MHC > type I-IIa MHC). The decline in the functional properties of single skeletal muscle fibers in the older animals appears to be more pronounced than what has been reported in younger animal populations.

Firing rate of the lower motoneuron and contractile properties of its muscle fibers after upper motoneuron lesion in man.

We studied motor unit (MU) firing rate and contractile properties and myosin isoform composition of single muscle fibers after upper motoneuron lesion. Single-MUs and surface electromyogram (EMG) were recorded during voluntary contractions and locomotion in the paretic (P) and nonparetic (NP) tibialis anterior (TA) of 15 hemiparetics. P TA low-threshold MUs fired within the lower end of their normal range. High-threshold MUs fired below their normal range or were not recruited. Surface EMG was abnormally low and high in the P TA and NP TA, respectively. On muscle cross sections stained with histochemical methods, type I fibers represented 99.4%, 74.3% and 66.6% of NP, P, and control TA, respectively. P TA fibers expressing type I myosin heavy chain (MyHC) were smaller, weaker, and slower. In conclusion, low MU firing rate and activity in the P TA was associated with slower type I MyHC fibers, while increased activity in NP TA resulted in homogenous expression of type I MyHC.

Modulation of MHC isoforms in functionally overloaded and exercised rat plantaris fibers.

The effects of 1 and 10 wk of functional overload (FO) of the rat plantaris with (FOTr) and without daily endurance treadmill training on its myosin heavy chain (MHC) composition were studied. After 1 and 10 wk of FO, plantaris mass was 22 and 56% greater in FO and 37 and 94% greater, respectively, in FOTr rats compared with age-matched controls. At 1 wk, pure type I and pure type IIa MHC fibers were hypertrophied in FO (39 and 44%) and FOTr (70 and 87%) rats. By 10 wk all fiber types comprising >5% of the fibers sampled showed a hypertrophic response in both FO groups. One week of FO increased the percentage of hybrid (containing both type I and type IIa MHC) fibers and of fibers containing embryonic MHC. By 10 wk, the percentage of pure type I MHC fibers was approximately 40% in both FO groups compared with 15% in controls, and the percentage of fibers containing embryonic MHC was similar to that in controls. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses showed an increase in type I MHC and a decrease in type IIb MHC in both FO groups at 10 wk, whereas little change was observed at 1 wk. These data are consistent with hypertrophy and transformation from faster to slower MHC isoforms in chronically overloaded muscles. The additional overload imposed by daily endurance treadmill training employed in this study (1.6 km/day; 10% incline) results in a larger hypertrophic response but appears to have a minimal effect on the MHC adaptations.

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 thyroid hormone (3,5,3'-triiodothyronine, T3) treatment on skeletal muscles of young (3-6 months) male Wistar rats were investigated in the present study. In the slow-twitch soleus, contraction and half-relaxation times of the isometric twitch were significantly shorter in hyperthyroid rats than in the control group, and twitch duration was shorter in rats treated with T3 for 8 weeks than for 4 weeks. All single soleus muscle fibres from hyperthyroid rats co-expressed types I and IIA myosin heavy chains (type I/IIA fibres) or type I, IIA and IIX myosin heavy chains (type I/IIAX fibres), while only type I MyHC fibres were isolated from the controls. A significantly higher content of type IIA myosin heavy chain and fast myosin light chain isoforms was observed in soleus fibres from the 8-week than from 4-week T3 group. There was no significant difference in maximum velocity of unloaded shortening (V0) between type I myosin heavy chain fibres from 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 length s-1, n = 33) and 8-week (1.03 +/- 0.31 muscle lengths s(-1), n = 31) groups, but type I/IIAX fibres from the 8-week T3 group had significantly higher V0 (1.56 +/- 0.10, n = 5) than type I from control and type I/IIA from hyperthyroid rats. In the fast-twitch extensor digitorum longus, neither myosin isoform composition, twitch duration nor V0 was affected by 4 or 8 weeks of T3 exposure. In conclusion, a dramatic and exposure duration-dependent change in the contractile speed of the isometric twitch and the expression of fast myosin isoforms was observed in soleus, but not in extensor digitorum longus, in response to T3 treatment. Long-term T3 treatment had relatively less influence, however, on V0 at the single cell level in spite of the dramatic increase in fast myosin isoforms.

Skeletal myoblast transplantation for repair of myocardial necrosis.

Myocardial infarcts heal by scarring because myocardium cannot regenerate. To determine if skeletal myoblasts could establish new contractile tissue, hearts of adult inbred rats were injured by freeze-thaw, and 3-4.5 x 10(6) neonatal skeletal muscle cells were transplanted immediately thereafter. At 1 d the graft cells were proliferating and did not express myosin heavy chain (MHC). By 3 d, multinucleated myotubes were present which expressed both embryonic and fast fiber MHCs. At 2 wk, electron microscopy demonstrated possible satellite stem cells. By 7 wk the grafts began expressing beta-MHC, a hallmark of the slow fiber phenotype; coexpression of embryonic, fast, and beta-MHC continued through 3 mo. Transplanting myoblasts 1 wk after injury yielded comparable results, except that grafts expressed beta-MHC sooner (by 2 wk). Grafts never expressed cardiac-specific MHC-alpha. Wounds containing 2-wk-old myoblast grafts contracted when stimulated ex vivo, and high frequency stimulation induced tetanus. Furthermore, the grafts could perform a cardiac-like duty cycle, alternating tetanus and relaxation, for at least 6 min. Thus, skeletal myoblasts can establish new muscle tissue when grafted into injured hearts, and this muscle can contract when stimulated electrically. Because the grafts convert to fatigue-resistant, slow twitch fibers, this new muscle may be suited to a cardiac work load.

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 (V0), 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. V0 did not change significantly in response to bed rest when data from all muscle cells were pooled. In two of the subjects, however, V0 decreased (P < 0.01-0.001) in muscle cells expressing the beta/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.