Specific Myosin Heavy Chain Isoforms Research Articles

Tongue muscle contractile, fatigue, and fiber type properties in rats.

The intrinsic and extrinsic tongue muscles manipulate the position and shape of the tongue and are activated during many oral and respiratory behaviors. In the present study, in 6-mo-old Fischer 344 rats, we examined mechanical and fatigue properties of tongue muscles in relation to their fiber type composition. In an ex vivo preparation, isometric force and fatigue was assessed by direct muscle stimulation. Tongue muscles were frozen in melting isopentane and transverse sections cut at 10 µm. In hematoxylin-eosin (H&E)-stained muscle sections, the relative fractions of muscle versus extracellular matrix were determined. Muscle fibers were classified as type I, IIa and IIx, and/or IIb based on immunoreactivity to specific myosin heavy chain isoform antibodies. Cross-sectional areas (CSAs) and proportions of different fiber types were used to calculate their relative contribution to total muscle CSAs. We found that the superior and inferior longitudinal intrinsic muscles (4.4 N/cm2) and genioglossus muscle (3.0 N/cm2) generated the greatest maximum isometric force compared with the transversalis muscle (0.9 N/cm2). The longitudinal muscles and the transversalis muscle displayed greater fatigue during repetitive stimulation consistent with the greater relative contribution of type IIx and/or IIb fibers. By contrast, the genioglossus, comprising a higher proportion of type I and IIa fibers, was more fatigue resistant. This study advances our understanding of the force, fatigue, and fiber type-specific properties of individual tongue musculature. The assessments and approach provide a readily accessible muscular readout for scenarios where motor control dysfunction or tongue weakness is evident.NEW & NOTEWORTHY For the individual tongue muscles, relatively little quantification of uniaxial force, fatigue, and fiber type-specific properties has been documented. Here, we assessed uniaxial-specific force generation, fatigability, and muscle fiber type-specific properties in the superior and inferior longitudinal muscles, the transversalis, and the genioglossus in Fischer 344 rats. The longitudinal muscles produced the greatest isometric tetanic-specific forces. The genioglossus was more fatigue resistant and comprised higher proportions of I and IIa fibers.

Myosin Heavy Chain Composition, Creatine Analogues, and the Relationship of Muscle Creatine Content and Fast-Twitch Proportion to Wilks Coefficient in Powerlifters.

Machek, SB, Hwang, PS, Cardaci, TD, Wilburn, DT, Bagley, JR, Blake, DT, Galpin, AJ, and Willoughby, DS. Myosin heavy chain composition, creatine analogues, and the relationship of muscle creatine content and fast-twitch proportion to Wilks coefficient in powerlifters. J Strength Cond Res 34(11): 3022-3030, 2020-Little data exist on powerlifting-specific skeletal muscle adaptations, and none elucidate sex differences in powerlifters. Powerlifters tend to display higher fast-twitch fiber content and phosphagen system dependence. Nevertheless, it is unknown whether fast-twitch fiber or muscle creatine content are predictive of competitive powerlifting performance (via Wilks coefficient). Twelve actively competing powerlifters (PL; n = 6M/6F; age = 21.3 ± 1.0; 3.0 ± 1.8 year competing; 7.3 ± 6.6 meets attended) and 10 sedentary controls (CON; n = 5M/5F; age = 19.4 ± 2.0 year) underwent vastus lateralis muscle biopsies and venipuncture to compare the myosin heavy chain (MHC) fiber type and creatine analogue profiles between groups of both sexes, and determine whether MHC IIa and muscle total creatine (MTC) composition predict powerlifting performance. Samples were analyzed for specific MHC isoform (I, IIa, and IIx) content via mixed homogenate SDS-PAGE, and creatine analogues (MTC, muscle creatine transporter [SLC6A8], serum total creatine [STC], and serum creatinine [CRT]). Furthermore, MHC IIa and MTC content were compared with Wilks coefficient using Pearson correlation coefficients. Male PL MHC content was 50 ± 6% I, 45 ± 6% IIa, and 5 ± 11% IIx, versus 46 ± 6% I, 53 ± 6 IIa, and 0% IIx in female PL. Conversely, male CON MHC content was 33 ± 5% I, 38 ± 7% IIa, and 30 ± 8% IIx, vs. 35 ± 9% I, 44 ± 8% IIa, and 21 ± 17% IIx in female CON. Muscle total creatine, SLC6A8, STC, and CRT did not significantly differ between groups nor sexes. Finally, neither MHC IIa content (r = -0.288; p = 0.364) nor MTC (r = 0.488; p = 0.108) significantly predicted Wilks coefficient, suggesting these characteristics alone do not determine powerlifting skill variation.

Prolongation of Relaxation Time in Extraocular Muscles With Brain Derived Neurotrophic Factor in Adult Rabbit.

PurposeWe tested the hypothesis that short-term treatment with brain derived neurotrophic factor (BDNF) would alter the contractile characteristics of rabbit extraocular muscle (EOM).MethodsOne week after injections of BDNF in adult rabbit superior rectus muscles, twitch properties were determined in treated and control muscles in vitro. Muscles were also examined for changes in mean cross-sectional areas, neuromuscular junction size, and percent of myofibers expressing specific myosin heavy chain isoforms, and sarcoendoplasmic reticulum calcium ATPases (SERCA) 1 and 2.ResultsBrain derived neurotrophic factor–treated muscles had prolonged relaxation times compared with control muscles. Time to 50% relaxation, time to 100% relaxation, and maximum rate of relaxation were increased by 24%, 27%, and 25%, respectively. No significant differences were seen in time to peak force, twitch force, or maximum rate of contraction. Brain derived neurotrophic factor treatment significantly increased mean cross-sectional areas of slow twitch and tonic myofibers, with increased areas ranging from 54% to 146%. Brain derived neurotrophic factor also resulted in an increased percentage of slow twitch myofibers in the orbital layers, ranging from 54% to 77%, and slow-tonic myofibers, ranging from 44% to 62%. No significant changes were seen SERCA1 or 2 expression or in neuromuscular junction size.ConclusionsShort-term treatment with BDNF significantly prolonged the duration and rate of relaxation time and increased expression of both slow-twitch and slow-tonic myosin-expressing myofibers without changes in neuromuscular junctions or SERCA expression. The changes induced by BDNF treatment might have potential therapeutic value in dampening/reducing uncontrolled eye oscillations in nystagmus.

Electrophoretic separation of reptilian skeletal and cardiac muscle myosin heavy chain isoforms: dependence on gel format.

This report provides a comparison of multiple gel formats to study myosin heavy chain (MHC) isoforms that are expressed in reptilian skeletal and cardiac muscles of five turtle species, water monitor, and prehensile tailed skink. Three gel formats were tested. The results identify one format that is superior, for the overall extent of electrophoretic separation and for the assessment of the number of MHC isoforms in reptilian striated muscles. The same format was shown previously to separate MHC isoforms that are expressed in American alligator. The results also show that another gel format reveals the distinct electrophoretic mobility of MHC isoforms in atrial, ventricular, and jaw adductor samples, compared to those expressed in skeletal muscles in the limbs and elsewhere in the body. In addition, the results reveal that the electrophoretic mobility of specific MHC isoforms, relative to other isoforms, depends on the gel format, as shown previously for mammalian and avian species. The discovery of the expression of masticatory MHC, which is abundantly expressed in jaw adductors of members of Carnivora and several other vertebrate orders, in the homologous muscles of prehensile tailed skink, an herbivore, and the carnivorous water monitor, was made during the course of this study.

Determination of Muscle Fiber Type in Rodents

Skeletal muscles consist of muscle fibers that can differ in both composition and functional characteristics. These three types of muscle fibers, broadly categorized as slow, fast IIa, and fast IIb muscle fibers, express characteristic myosin heavy chain proteins and have different metabolic and enzymatic activities, which can be used as surrogate markers to identify the different fiber types. Pathological changes affecting the muscle, such as denervation, muscle disuse, and atrophy not only manifest on a functional level, but also as marked changes in the composition of muscle fiber type of individual muscles. In this unit we describe three methods for histological identification of slow/type I, fast fatigue resistant/type IIa, and fast fatigable/type IIb fibers by staining for either myosin ATPases or oxidative enzyme capacity (succinate dehydrogenase, SDH)-or, alternatively, immunostaining for specific myosin heavy chain isoforms in muscles of mouse hindlimbs. Curr. Protoc. Mouse Biol. 2:231-243 © 2012 by John Wiley & Sons, Inc.

Speed and endurance: you can have it all

skeletal muscle comprises multiple fiber types that vary in their contractile and metabolic properties. Muscle fibers are classified primarily on the basis of the specific myosin heavy chain isoform that they express. Slow twitch or type I fibers are rich in mitochondria, rely principally on

Stapedius muscle fibre characterization in the noise exposed and auditory deprived rat

In skeletal muscle, interventions that unload the muscle cause slow-to-fast myosin heavy chain (MHC) conversions, whereas fast-to-slow conversions are seen when the muscles are engaged in resistance training and endurance exercise. The stapedius muscle (SM) is reported to prevent cochlear damage by noise. This theory may be supported by showing comparable changes of muscle fibre composition when ears are exposed to longstanding noise (SM training). Comparable changes after sound deprivation (SM unloading) would suggest that the SM needs a certain degree of daily activity evoked by environmental sound to sustain its normal composition. We investigated the difference in myosin composition of SM fibres from rats exposed to noise, from auditory deprived rats and from rats exposed to low level ambient noise (control group). Consecutive complete SM cross-sections were processed by enzymehistochemistry to determine acid/alkali lability of myofibrillar adenosine triphosphatase (mATPase) and by immunohistochemistry using MHC antibodies. Fibres were assigned to mATPase type I, IIA, IIX or ‘Miscellaneous’ categories. Per mATPase category, the fibres were attributed to groups with specific MHC isoform compositions. Auditory deprivation lasting nine weeks was accomplished by closure of the external meatus at the age of three weeks. A slow-to-fast shift was seen in these rats when compared to the control group. The noise exposed group was exposed to 65–90 dB sound pressure level during a period lasting nine weeks from the age of three weeks onwards. A shift from an overwhelming presence of type mATPase IIX, as seen in the control group, to type mATPase IIA occurred in the noise exposed group. Also, more MHC IIA/IIX hybrid fibres were found in the mATPase IIX category. An adaptive response to the acoustic environment in the characteristics of the fibres of the SM, comparable to the response in skeletal muscles on unloading and training activity, can be ascertained. This supports the theory that the SM plays an active role in modulating external acoustic energy on entry to the cochlea. Our results are also in favour of another postulated function of the SM, the unmasking of high-frequency signals in low-frequency background noise.

Highlights From The Literature

Highlights From The Literature

Stapedius muscle fiber characterization during postnatal development in the rat

The stapedius muscle (SM) is reported to prevent cochlear damage by noise. Functional demands are then the ability of fast contraction with long endurance. At the end of the third postnatal week, the middle ear of the rat is completely pneumatized and according to electrophysiological data, the auditory function starts to match the adult. We investigated the developmental changes in myosin composition of SM fibres using consecutive complete SM cross-sections (taken from rats on post natal day (PND) 7, 14, 16, 21, 28, 42 and 84) which were processed by enzymehistochemistry to determine acid/alkali lability of myofibrillar adenosine triphosphatase (mATPase) and by immunohistochemistry using myosin heavy chain (MHC) antibodies (mAb). Fibres were assigned to mATPase type I, IIA, IIB, IIX or ‘Miscellaneous’ categories. Per mATPase category, the fibres were attributed to groups with specific MHC isoform compositions. Neonatal MHC expression could not be documented with the mAb used. However, embryonal (Emb) MHC was expressed at PND 7, very little at PND 14; at later PND fibres did not show Emb MHC. In general, the mATPase-based classification did not show large alterations after PND 21. Expression of MHC IIB, which was present in almost 50% of the fibres at PND 7 and 14, diminished to 3% at PND 84. A decrease in number of fibres expressing more than one MHC isoform was found. These results show that the SM is a precociously developing muscle compared to limb muscles and even to the diaphragm. Moreover, it is shown that the expression of the adult MHC isoform phenotype coincides with the onset of auditory function in the third postnatal week.

Surface electromyography and peak torque of repetitive maximum isokinetic plantar flexions in relation to aspects of muscle morphology

This study investigates the relationships between surface electromyography (EMG [Mean frequency of the power spectrum (MNF)]) and peak torque variables obtained during 100 maximum concentric plantar flexions with the right limb at 60° s −1 and different muscle morphological variables. Surface EMG was recorded from the right gastrocnemius lateralis and muscle biopsies were taken from the same site as the EMG electrodes were positioned. Muscle fibre area and fibre type composition were determined on serial muscle cross sections using both histochemistry (myofibrillar adenosine triphosphatase) and immunohistochemistry (monoclonal antibodies against specific myosin heavy chain isoforms). Forty-three female and nine male students participated in the study. Gastrocnemius lateralis contained predominantly type I fibres (50%) and type IIA fibres (40%) in both sexes and large individual differences were found. Principal component analysis (PCA) was used for the intercorrelation analyses, and projection to latent structures (PLS) was used for the multivariate regression analysis. MNF correlated positively with different fibre areas and with the proportion of type I fibres. Fibre areas and sex were the most important factors in the regression of maximum peak torque. High proportion of type I fibres and sex were the most important regressors of peak torque endurance normalised for lean body mass. More studies are needed to understand the complex interrelationships between intrinsic muscle properties and the frequency content of the surface EMG before theoretical models can be formulated that incorporate both fibre areas and fibre type proportions.

Effects of thyroid hormone receptor gene disruption on myosin isoform expression in mouse skeletal muscles.

Skeletal muscle is known to be a target for the active metabolite of thyroid hormone, i.e., 3,5,3'-triiodothyronine (T(3)). T(3) acts by repressing or activating genes coding for different myosin heavy chain (MHC) isoforms via T(3) receptors (TRs). The diverse function of T(3) is presumed to be mediated by TR-alpha(1) and TR-beta, but the function of specific TRs in regulating MHC isoform expression has remained undefined. In this study, TR-deficient mice were used to expand our knowledge of the mechanisms by which T(3) regulates the expression of specific MHC isoforms via distinct TRs. In fast-twitch extensor digitorum longus (EDL) muscle, TR-alpha(1)-, TR-beta-, or TR-alpha(1)beta-deficient mice showed a small but statistically significant decrease (P < 0.05) of type IIB MHC content and an increased number of type I fibers. In the slow-twitch soleus, the beta/slow MHC (type I) isoform was significantly (P < 0. 001) upregulated in the TR-deficient mice, but this effect was highly dependent on the type of receptor deleted. The lack of TR-beta had no significant effect on the expression of MHC isoforms. An increase (P < 0.05) of type I MHC was observed in the TR-alpha(1)-deficient muscle. A dramatic overexpression (P < 0.001) of the slow type I MHC and a corresponding downregulation of the fast type IIA MHC (P < 0.001) was observed in TR-alpha(1)beta-deficient mice. The muscle- and fiber-specific differences in MHC isoform expression in the TR-alpha(1)beta-deficient mice resembled the MHC isoform transitions reported in hypothyroid animals, i.e., a mild MHC transition in the EDL, a dramatic but not complete upregulation of the beta/slow MHC isoform in the soleus, and a variable response to TR deficiency in different soleus muscle fibers. Thus the consequences on muscle are similar in the absence of thyroid hormone or absence of thyroid hormone receptors, indicating that TR-alpha(1) and TR-beta together mediate the known actions of T(3). However, it remains unknown how thyroid hormone exerts muscle- and muscle fiber-specific effects in its action. Finally, although developmental MHC transitions were not studied specifically in this study, the absence of embryonic and fetal MHC isoforms in the TR-deficient mice indicates that ultimately the transition to the adult MHC isoforms is not solely mediated by TRs.

The impact of biochemical methods for single muscle fibre analysis.

Biochemical methods for single muscle fibre analysis provide sensitive measures for elucidating muscle fibre heterogeneity. The understanding of the complexity of skeletal muscle fibres, initially based on qualitative histochemistry and immunohistochemistry, has been greatly expanded by quantitative micromethods, such as microphotometry and microbiochemical assays. Assessment of metabolic enzyme activity levels has revealed pronounced scattering within and between different fibre types and has highlighted the use of specific enzyme activity ratios as discriminative measures. With the exception of type I fibres, metabolic properties are loosely coupled with molecular properties of the myofibrillar apparatus. As such, myosin heavy chain (MHC) isoforms appear to be the best choice for fibre type delineation. Among the two available methods for MHC-based fibre type distinction, single fibre electrophoresis appears to be superior to immunohistochemistry. The electrophoretic separation of MHC isoforms in single fibres is quantitative and, as opposed to immunohistochemistry, yields important information on MHC isoform proportions in hybrid fibres. Histochemical staining for myofibrillar ATPase activity can, thus, be correlated in most cases with specific MHC isoform profiles. Single fibre studies have demonstrated a relationship between ATP phosphorylation potential and MHC isoform complement. This relationship corresponds to different tension costs and provides an additional rationale for the MHC-based fibre type diversity and transitions. The combination of reverse transcriptase (RT) with polymerase chain reaction (PCR) has proved to be a highly sensitive tool and has extended single fibre analysis to the level of MHC mRNA isoforms. Application of RT-PCR techniques to single fibre fragments identified by their MHC protein isoform profile, provides insights at two levels of expression and, thus, has extended our knowledge on the plasticity of muscle and the dynamical state of muscle fibres.

Immunohistochemical Classification of Skeletal Muscle Fibers.

Mammalian skeletal muscle fibers have several types of myosin heavy chain (MHC) isoforms. Immunohistochemical techniques enable us to identify the MHC isoform type in a single muscle fiber, i.e., MHCI, MHCIIa, MHCIIx, or MHCIIb. In addition, hybrid single fibers having more than one MHC isoform type can be classified using immunohistochemistry. The presence of different MHC isoform(s) in a single fiber correlates with the maximal contractile velocity and the enzyme histochemical reaction of the myofibrillar adenosine triphosphatase. The adaptive response of the skeletal muscle is the basis of the fiber-type transformation depending on the phenotypic expression pattern in several MHC isoforms. In this review article, we address the availability of the monoclonal antibodies which recognize specific MHC isoform(s) in a single fiber.

Human skeletal muscle fiber types: delineation, development, and distribution.

This brief review attempts to summarize a number of studies on the delineation, development, and distribution of human skeletal muscle fiber types. A total of seven fiber types can be identified in human limb and trunk musculature based on the pH stability/lability of myofibrillar adenosine triphosphatase (mATPase). For most human muscles, mATPase-based fiber types correlate with the myosin heavy chain (MHC) content. Thus, each histochemically identified fiber has a specific MHC profile. Although this categorization is useful, it must be realized that muscle fibers are highly adaptable and that innumerable fiber type transients exist. Also, some muscles contain specific MHC isoforms and/or combinations that do not permit routine mATPase-based fiber typing. Although the major populations of fast and slow are, for the most part, established shortly after birth, subtle alterations take place throughout life. These changes appear to relate to alterations in activity and/or hormonal levels, and perhaps later in life, total fiber number. Because large variations in fiber type distribution can be found within a muscle and between individuals, interpretation of data gathered from human muscle is often difficult.

Correlation between myofibrillar ATPase activity and myosin heavy chain composition in equine skeletal muscle and the influence of training.

The histochemical myofibrillar ATPase (mATPase) method is used routinely for identification of equine skeletal muscle fiber types, but important problems have been observed with the subdivision of fast fiber population when using this method. To verify the use of this qualitative method, a number of equine muscle biopsies were analyzed with a combination of histochemical, immunohistochemical, electrophoretic, and morphometric techniques. The influence of training on these interrelations was also evaluated. Five young (2-3 years old) thoroughbred horses were intensively trained for 8 months on a high-speed treadmill. Biopsies were taken from the gluteus medius muscle at the beginning, after 4 months, and at the end of the training program. Serial sections of the samples were stained by mATPase histochemistry and immunohistochemistry by using a number of monoclonal antibodies specific to selected myosin heavy chain (MyHC) isoforms. The histochemical and immunohistochemical categorization of a large number of fibers (N = 2,078) was compared fiber by fiber. The MyHC content of homogenates of the same biopsies were quantified by densitometry of a sensitive gel electrophoretic technique and compared with histochemical and immunohistochemical fiber types. A large proportion of fibers examined (approximately 20%) were misclassified by traditional mATPase histochemistry. Many fibers histochemically identified as type IIB displayed both type IIa and type IIb MyHC isoforms, and nearly all type IIAB fibers in mATPase contained only the type IIa MyHC isoform by immunohistochemistry. Correlation analyses suggested a weak relation between the histochemically assessed relative cross-sectional area occupied by the three major fiber types (I, IIA, and IIB) and the electrophoretically assessed MyHC content, whereas a stronger relation was found between immunohistochemically defined fiber types and electrophoretic data. The four fiber type populations delineated according to MyHC content (I, IIA, IIAB, and IIB) had sizes and oxidative capacities significantly different from each other. No adaptation of any parameter measured to training was found. Training had no significant effect on the number of fibers misclassified by mATPase histochemistry. These data demonstrate a significant limitation in mATPase histochemistry for assessing fibers containing fast MyHC isoforms. The use of monoclonal antibodies against specific MyHC isoforms seems to be a more sensitive and less subjective method.

Developmentally regulated neural protein EAP-300 is expressed by myocardium and cardiac neural crest during chick embryogenesis.

The spatiotemporal distribution of EAP-300 (embryonic avian polypeptide of 300 kDa) was analyzed in embryonic chick heart using immunohistochemistry and confocal microscopy. EAP-300 is a developmentally regulated protein initially characterized in neural cells from chick retina. Myocardial cells all along the early tubular heart were ubiquitously immunolabeled for EAP-300 by embryonic day 2 (E2, Stage 13)). At E5 (Stage 24), myocardial EAP-300 expression levels remained significant in both atrial and ventricular myocardium. At E6 (Stage 28), distinct populations of EAP-300 immunolabeled cells were also observed external to the heart, in septal mesenchymal tissue and neural ganglia adjacent to the outflow tract; these cell populations were confirmed as neural crest-derived by co-localization of EAP-300 and HNK-1. At E13 (Stage 39), myocardial immunolabeling for EAP-300 was no longer ubiquitous, but increasingly restricted to conduction tissues, including the atrioventricular bundle and subendocardial Purkinje cells. This restriction of immunolabeling could be demonstrated definitively at E15 (stage 41), by which stage subendocardial and periarterial Purkinje fibers were clearly immunoreactive for EAP-300 and several known markers of chick conduction tissue, including specific myosin heavy chain isoforms and connexin42, a gap junctional protein preferentially expressed by Purkinje fibers. Just prior to hatching at E21 (Stage 46), immunolabeling of conduction tissues was reduced, although still above that of non-conductile myocardium. This spatiotemporal map of cardiac EAP-300 expression indicates that it is independently and transiently expressed in early myocardium, cardiac conduction tissue, and neural crest derivatives during development.

Sequence requirements for myosin gene expression and regulation in Caenorhabditis elegans.

Four Caenorhabditis elegans genes encode muscle-type specific myosin heavy chain isoforms: myo-1 and myo-2 are expressed in the pharyngeal muscles; unc-54 and myo-3 are expressed in body wall muscles. We have used transformation-rescue and lacZ fusion assays to determine sequence requirements for regulated myosin gene expression during development. Multiple tissue-specific activation elements are present for all four genes. For each of the four genes, sequences upstream of the coding region are tissue-specific promoters, as shown by their ability to drive expression of a reporter gene (lacZ) in the appropriate muscle type. Each gene contains at least one additional tissue-specific regulatory element, as defined by the ability to enhance expression of a heterologous promoter in the appropriate muscle type. In rescue experiments with unc-54, two further requirements apparently independent of tissue specificity were found: sequences within the 3' non-coding region are essential for activity while an intron near the 5' end augments expression levels. The general intron stimulation is apparently independent of intron sequence, indicating a mechanistic effect of splicing. To further characterize the myosin gene promoters and to examine the types of enhancer sequences in the genome, we have initiated a screen of C. elegans genomic DNA for fragments capable of enhancing the myo-2 promoter. The properties of enhancers recovered from this screen suggest that the promoter is limited to muscle cells in its ability to respond to enhancers.

Three myosin heavy chain isoforms in type 2 skeletal muscle fibres

Mammalian skeletal muscles consist of three main fibre types, type 1,2A and 2B fibres, with different myosin heavy chain (MHC) composition. We have now identified another fibre type, called type 2X fibre, characterized by a specific MHC isoform. Type 2X fibres, which are widely distributed in rat skeletal muscles, can be distinguished from 2A and 2B fibres by histochemical ATPase activity and by their unique staining pattern with seven anti-MHC monoclonal antibodies. The existence of the 2X-MHC isoform was confirmed by immunoblotting analysis using muscles containing 2X fibres as a major component, such as the normal and hyperthyroid diaphragm, and the soleus muscle after high frequency chronic stimulation. 2X-MHC contains one determinant common to 2B-MHC and another common to all type 2-MHCs, but lacks epitopes specific for 2A- and 2B-MHCs, as well as an epitope present on all other MHCs. By SDS-polyacrylamide gel electrophoresis 2X-MHC shows a lower mobility compared to 2B-MHC and appears to comigrate with 2A-MHC. Muscles containing predominantly 2X-MHC display a velocity of shortening intermediate between that of slow muscles and that of fast muscles composed predominantly of 2B fibres.