Myosin Heavy Chain Isoform Expression Research Articles

Myosin heavy chain isoform transitions in canine skeletal muscles during postnatal growth

To gain a better understanding of the normal characteristics of developing canine muscles, myosin heavy chain (MHC) isoform expression was analysed in the axial and limb skeletal muscles of 18 young dogs whose ages ranged from the late prenatal stage to 6 months. We compared the results of immunohistochemistry using ten monoclonal antibodies, specific to different MHC isoforms, and enzyme-histochemical reactions, which demonstrate the activity of myofibrillar ATPase, succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (alpha-GPDH). In the skeletal muscles of fetuses and neonatal dogs the developmental isoforms MHC-emb and MHC-neo were prevalent. In all muscles the primary fibres, located centrally in each muscle fascicle, strongly expressed the slow isoform MHC-I. The adult fast isoform MHC-IIa was first noted in some of the secondary fibres on fetal day 55. During the first 10 days after birth, the expression of MHC-emb declined, as did that of MHC-neo during the second and third weeks. Correspondingly, the expression of MHC-IIa, and later, of MHC-I increased in the secondary fibres. Between the sixth week and second month the expression of MHC-IIx became prominent. The slow rhomboideus muscle exhibited an early expression of the slow isoform in the secondary fibres. Our results indicate that the timing of muscle maturation depends on its activity immediately following birth. The fastest developing muscle was the diaphragm, followed by the fast muscles. A pronounced changeover from developmental to adult isoforms was noted at 4-6 weeks of age, which coincides with the increased physical activity of puppies.

The Impact of Mechanical Ventilation on Immature Airway Smooth Muscle: Functional, Structural, Histological, and Molecular Correlates

Preterm infants exposed to mechanical ventilation often develop airway dysfunction and bronchopulmonary dysplasia. The mechanisms of mechanical ventilation-induced airway injury are currently unknown. This study correlates the age-related effects of mechanical ventilation on airway function with structural alterations at the tissue, cellular, ultrastructural, and molecular levels. Mechanically ventilated and nonventilated tracheal rings were obtained from premature and newborn lambs. In tissue baths, the passive and active length-tension relationships and dose-response characteristics of the tracheal rings were determined. Fixed tracheal rings were digested and the resulting isolated smooth muscle cells measured. Rings were analyzed by light and electron microscopy. Additionally, protein was extracted from the tracheal smooth muscle and myosin heavy chain isoforms were separated by SDS-polyacrylamide gel electrophoresis and analyzed by densitometry. Mechanical ventilation resulted in a significant decrease of both the slope of the passive length-stress relationship and of maximal force generation, with both effects being most pronounced in the newborn age group. These age-related functional alterations correlated with a decrease in smooth muscle cell length and a disruption of ultrastructural architecture, which were also most pronounced in the older groups. Furthermore, mechanical ventilation resulted in epithelial denudation at all ages. There were no acute statistically significant effects of mechanical ventilation on myosin heavy chain isoform expression. This study demonstrates age-related effects of mechanical ventilation on the passive and active characteristics of tracheal function and provides a structural analysis of potential mechanisms. The mechanisms behind these functional differences involve ultrastructural changes in cell length, tissue matrix, and disruption of epithelial integrity. These findings help elucidate the pathogenesis of ventilator-induced airway injury.

Effects of a Nerve‐Muscle Pedicle on the Denervated Rat Thyroarytenoid Muscle

To evaluate the effects of the nerve-muscle pedicle (NMP) method on the rat thyroarytenoid (TA) muscle after transection of the recurrent laryngeal nerve (RLN). Quantitative histologic assessment of the TA muscle after NMP implantation. Thirty-six Wistar rats were divided into two groups: animals subjected to transection of the left RLN alone (DNV group) and animals subjected to transection of the left RLN followed by immediate transplantation of a NMP flap containing the sternohyoid (SH) muscle and ansa cervicalis nerve branch (NMP group). Animals were killed 2, 4, and 10 weeks after the treatments. The TA muscle stained with hematoxylin-eosin was evaluated quantitatively. The pre- and postsynaptic structures of the neuromuscular junction (NMJ) in the TA muscle were analyzed histochemically. The myosin heavy chain (MyHC) isoforms were evaluated using immunohistochemistry and reverse-transcriptase polymerase chain reaction (RT-PCR). In the NMP group, the TA muscle fiber recovered to almost normal at 10 weeks, and the ratio of the number of synaptophysin-positive nerve terminals to that of alpha-bungarotoxin-positive acetylcholine receptors recovered to 79.8 +/- 11.8% (P < .05, compared with the control). Immunohistochemistry and the RT-PCR method after laser capture microdissection revealed the expression of MyHC isoforms type 2B and type 2A; the latter was detected in the SH muscle but not in the normal or denervated TA muscle. The NMP method was effective for recovering from the atrophic changes of the TA muscle after transection of the RLN. This was attributed to successful reinnervation by reconstruction of the NMJ, which might change MyHC isoform expression.

Regional differences in myosin heavy chain isoform expression and maximal shortening velocity of the rat vaginal wall smooth muscle

Contractility of the proximal and distal vaginal wall smooth muscle may play distinct roles in the female sexual response and pelvic support. The goal of this study was to determine whether differences in contractile characteristics of smooth muscle from these regions reside in differences in the expression of isoforms of myosin, the molecular motor for muscle contraction. Adult female Sprague-Dawley rats were killed on the day of estrus, and the vagina was dissected into proximal and distal segments. The Vmax at peak force was greater for tissue strips of the proximal vagina compared with that of distal (P < 0.01), although, at steady state, the Vmax for the muscle strips from the two regions was not different. Furthermore, at steady state, muscle stress was higher (P < 0.001) for distal vaginal strips (n = 5). Consistent with the high Vmax for the proximal vaginal strips, RT-PCR results revealed a higher %SM-B (P < 0.001) in the proximal vagina. A greater expression of SM-B protein (P < 0.001) was also detected by Western blotting (n = 4). Interestingly, there was no regional difference noted in SM-1/SM-2 isoforms (n = 6). The proximal vagina had a higher expression of myosin heavy chain protein (P < 0.01) and a greater percentage of smooth muscle bundles (P < 0.001). The results of this study are the first demonstration of a regional heterogeneity in Vmax and myosin isoform distribution in the vagina wall smooth muscle and confirm that the proximal vaginal smooth muscle exhibits phasic contractile characteristics compared with the distal vaginal smooth muscle, which is tonic.

Effects of hindlimb unweighting and aging on rat semimembranosus muscle and myosin

We tested the hypothesis that lower specific force (force/cross-sectional area) generated by type II fibers from hindlimb-unweighted rats resulted from structural changes in myosin (i.e., a change in the ratio of myosin cross bridges in the weak- and strong-binding state during contraction). In addition, we determined whether those changes were age dependent. Permeabilized semimembranosus muscle fibers from young adult and aged rats, some of which were hindlimb unweighted for 3 wk, were studied for Ca(2+)-activated force generation and maximal unloaded shortening velocity. Fibers were also spin labeled specifically at myosin Cys707 to assess the structural distribution of myosin during maximal isometric contraction using electron paramagnetic resonance spectroscopy. Myosin heavy chain isoform (MHC) expression and the ratio of MHC to actin were evaluated in each fiber. Fibers from the unweighted rats generated 34% less specific force than fibers from weight-bearing rats (P < 0.001), independent of age. Electron paramagnetic resonance analyses showed that the fraction of myosin heads in the strong-binding structural state during contraction was 11% lower in fibers from the unweighted rats (P = 0.019), independent of age. More fibers from unweighted rats coexpressed MHC IIB-IIX compared with fibers from weight-bearing rats (P = 0.049). Unweighting induced a slowing of maximal unloaded shortening velocity and an increase in the ratio of MHC to actin in fibers from young rats only. These data indicate that altered myosin structural distribution during contraction and a preferential loss of actin contribute to unweighting-induced muscle weakness. Furthermore, the age of the rat has an influence on some parameters of changes in muscle contractility that are induced by unweighting.

GATA4 and the Two Sides of Gene Expression Reprogramming

See related article, pages 837–845 During embryonic development as well as in postnatal life, cardiac cells are continuously exposed to a host of extracellular signals to which they adapt and respond by activating intracellular signaling pathways and modulating the expression of nuclear genes. In the adult heart, the most common and easily recognized response to stressful stimuli is hypertrophy of cardiomyocytes and remodeling of the entire organ.1 This reactive growth response requires de novo synthesis of contractile and structural proteins, obtained through a complex series of events culminating in gene expression reprogramming.2 GATA4 is a zinc-finger transcription factor highly expressed in cardiomyocytes at different developmental stages where it regulates the transcription of several structural and regulatory genes, including atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), α- and β-myosin heavy chain (MHC).3 In human hearts, GATA4 has also been demonstrated to be a critical regulator of cardiac development, as shown by the association between GATA4 mutations and the presence of congenital cardiac malformations.4 The specific effects of GATA4 deletion during embryonic age have been extensively investigated in genetically modified mouse models.5–7 Mice homozygous for a GATA4 null allele6 or homozygous GATA4-deficient mice (GATA4−/−)5 die early in development because of abnormal …

SERCA isoform expression in paralyzed muscle

The influence of paralysis on the expression of sarco(endo)plasmic reticulum calcium ATPase (SERCA) pump isoforms in the rat soleus muscle was assessed. Paralysis was induced in rats via a complete mid-thoracic spinal cord transection (ST). SERCA pump protein isoform expression in single muscle fibers was evaluated at 15 and 30 days post-ST using immunohistochemistry and compared to myosin heavy chain (MyHC) isoform content. In control rats, ~60% of soleus fibers examined contained slow MyHC and SERCA isoforms, another ~25% contained slow MyHC and both fast and slow SERCA isoforms. The majority of the remaining fibers contained fast isoforms of MyHC and SERCA. Fifteen days after ST no fibers contained both slow MyHC and SERCA, ~60% contained slow MyHC and both SERCA isoforms, and ~25% contained fast and slow MyHC and SERCA isoforms. Thirty days after ST no fibers contained both slow MyHC and SERCA, ~30% contained slow MyHC and both SERCA isoforms, ~30% contained fast and slow MyHC and SERCA isoforms, ~20% contained both fast and slow MyHC and only fast SERCA isoforms, and ~20% contained just the fast isoforms of MyHC and SERCA. These data are consistent with a more rapid slow to fast transformation in SERCA than MyHC isoforms after paralysis and suggest that the cellular factors that direct transformations in SERCA isoform expression are distinct from those that direct transformations in MyHC isoform expression after paralysis. Supported by NIH GM53933.

Regional myosin heavy chain isoform expression in response to exercise training in old rat myocardium

It is recognized that with ageing of the rodent heart there is a shift in myosin heavy chain (MHC) isoform from α-MHC to β-MHC. Previous studies have shown evidence of significant regional differen...

The Effect of Resection on Satellite Cell Activity in Rabbit Extraocular Muscle

A common treatment for motility disorders of the extraocular muscles (EOMs) is a resection procedure in which there is surgical shortening of the muscle. This procedure results in rotation of the globe toward the resected muscle, increased resting tension across the agonist-antagonist pair, and stretching of the elastic components of the muscles. In the rabbit, due to orbital constraints and limited rotation, resection results in more significant stretch of the surgically treated muscle than the antagonist. This surgical preparation allows for the examination of the effects of surgical shortening of one rectus muscle and passive stretch of its ipsilateral antagonist. The insertional 6 mm of the superior rectus muscles of adult rabbits were resected and reattached to the original insertion site. After 7 and 14 days, the animals were injected intraperitoneally with bromodeoxyuridine (BrdU) every 2 hours for 12 hours, followed by a 24-hour BrdU-free period. All superior and inferior rectus muscles from both globes were examined for BrdU incorporation, MyoD expression, neonatal and developmental myosin heavy chain (MyHC) isoform expression, and myofiber cross-sectional area alterations. In the resected muscle and in the passively stretched antagonist muscle, there was a dramatic increase in the number of myofibers positive for neonatal MyHC and in the number of BrdU- and MyoD-positive satellite cells. The addition of BrdU-positive myonuclei increased from 1 per 1000 myofibers in cross sections of control muscles to 2 to 3 per 100 myofibers in the resected muscles. Single myofiber reconstructions showed that multiple BrdU-positive myonuclei were added to individual myofibers. Addition of new myonuclei occurred in random locations along the myofiber length of single fibers. There was no correlation between myofibers with BrdU-positive myonuclei and neonatal MyHC isoform expression. Both active and passive stretch of the rectus muscles produced by strabismus surgery dramatically upregulated the processes of satellite cell activation, integration of new myonuclei into existing myofibers, and concomitant upregulation of immature myosin heavy chain isoforms. Understanding the effects of strabismus surgery on muscle cell biological reactions and myofiber remodeling may suggest new approaches for improving surgical outcomes.

Denervation effects on myonuclear domain size of rat diaphragm fibers

Denervation (DNV) of rat diaphragm muscle (DIAm) leads to selective atrophy of type IIx and IIb fibers, whereas the cross-sectional area of type I and IIa fibers remains unchanged or slightly hypertrophied. DIAm DNV also increases satellite cell mitotic activity and myonuclear apoptosis. Similar to other skeletal muscles, DIAm fibers are multinucleated, and each myonucleus regulates the gene products in a finite fiber volume, i.e., myonuclear domain (MND). MND size varies across DIAm fiber types in rank order, I < IIa < IIx < IIb [fiber type based on myosin heavy chain isoform expression]. We hypothesized that, after DNV, the total number of myonuclei per fiber does not change and, accordingly, that MND changes proportionately to the change in fiber size regardless of fiber type. Adult rats underwent unilateral (right side) DIAm DNV, and after 2 wk single fibers were dissected. Fiber cross-sectional area, myonuclear number, and MND were measured by confocal microscopy, and these values in DNV DIAm were compared with those obtained in controls. After DNV, type I fibers hypertrophied, type IIa fiber size was unchanged, and type IIx and IIb fibers atrophied compared with control. The total number of myonuclei per fiber was not affected by DNV. Accordingly, after DNV, type I fiber MND increased by 25%, whereas it decreased in type IIx and IIb fibers by 50 and 70%, respectively. These results suggest that MND is not maintained after DNV-induced DIAm fiber hypertrophy or atrophy. These results are interpreted with respect to consequent effects of DNV on myonuclear transcriptional activity and protein turnover.

Levels of myosin heavy chain mRNA transcripts and content of protein isoforms in the slow soleus muscle of 7-month-old rats with altered thyroid status

Skeletal muscles of small rodents contain four main fiber types, namely type 1, 2A, 2X/D and 2B fibers containing myosin heavy chain (MyHC) 1, 2a, 2x/d and 2b isoforms. Each of these MyHC isoforms is the product of a distinct gene and their expression is believed to be primarily transcriptionally controlled. In most rat muscles, messenger RNA (mRNA) transcripts for MyHC1, 2a, 2x/d and 2b and their corresponding protein products were found with the exception of the soleus muscle, where typically only MyHC1 and 2a transcripts and protein isoforms were demonstrated under normal conditions. Here we show the expression of all four MyHC1, 2a, 2x/d and 2b mRNA transcripts in the soleus muscle under normal conditions in euthyroid, as well as in experimental hypothyroid and hyperthyroid (with the exception of 2b MyHC transcript) 7-month-old female inbred Lewis rats. This is not matched, however, by the appearance of corresponding four isoforms, as we have found that 2x/d and 2b protein isoforms are not present at levels detectable by SDS-PAGE. We also show that the chronic hypothyroid and hyperthyroid status affects the expression of MyHC isoforms both at the mRNA and protein levels.

Three-dimensional compartmentalization of myosin heavy chain and myosin light chain isoforms in dog thyroarytenoid muscle

The thyroarytenoid muscle, a vocal fold adductor, has important roles in airway protection (e.g., prevention of aspiration) and phonation. Isoform expression of myosin heavy chain (MHC), a major determinant of muscle-shortening velocity, has been reported to be heterogeneous in this muscle in several mammals, differing markedly between the medial and lateral divisions. The objective was to determine the isoform expression patterns of both MHC and myosin light chain (MLC), with the latter having a modulatory role in determining shortening velocity, to further test whether the expression of both myosin subunits differs in multiple specific sites within the divisions of the dog thyroarytenoid muscle, potentially revealing even greater compartmentalization in this muscle. Our results indicate the existence of large gradients in the relative levels of individual MHC isoforms in the craniocaudal axis along the medial layer (i.e., airflow axis), where levels of MHC-I and MHC-IIA are low at both ends of the axis and high in the middle and MHC-IIB has a reciprocal distribution. The lateral layer is more uniform, with high levels of MHC-IIB throughout. The level of MHC-IID is relatively constant along the axis in both layers. Large differences exist in the distribution of MHC isoforms among single fibers isolated from sites along the craniocaudal axis, especially in the lateral layer. Systematic regional variations are apparent in the MLC isoform composition of single fibers as well, including some MLC isoform combinations that are not observed in dog limb muscles. Variations of MHC and MLC isoform expression in the dog thyroarytenoid muscle are greater than previously recognized and suggest an even broader range of contractile properties within this multifunctional muscle.

Myosin Heavy Chain Transitions in Dog Skeletal Muscle Fibres during Post‐natal Development

Myosin heavy chain (MHC) isoform expression was established in the masseter, axial and limb skeletal muscles in 18 young dogs from late prenatal to 6 months of age. Results obtained by immunohistochemical procedure using 11 monoclonal antibodies, SDS‐PAGE electrophoresis, Western‐blotting and enzyme‐histochemical reactions were compared to those found in skeletal muscles of adult dogs. In the skeletal muscles of both fetuses in the last stages of development and neonatal dogs, the developmental isoforms MHC‐emb and MHC‐neo were prevalent. During the first 10 days after birth, the level of embryonic isoform expression declined, as did that of the neonatal isoform during the second and third weeks. Correspondingly, the expression of MHC‐IIa, and later, of MHC‐I increased. By 6 weeks of age the presence of MHC‐IIx became obvious. This transitional pattern was similar in all muscles except in the rhomboideus, where secondary slow fibres were present before birth. In the other muscles, only one primary, centrally located, slow fibre was present in a fascicle and the secondary slow fibres evolved from developmentally fast fibres between the second and third weeks of age. Our results indicate that timing of muscle maturation depends on its activity right after birth. The fastest developing muscles were the masseter muscle and the diaphragm. They were followed by others, postural and locomotory muscles. A pronounced swap from developmental to adult isoforms was noted between the 4th and 6th week of age. At this age puppies are weaned and begin playing intensively and investigating their surroundings. The results of our study indicate that dog skeletal muscles exhibit an immature morphology at birth and develop relatively slowly compared to those of other domestic animals.

Myosin Heavy Chain Isoforms in Equine Gluteus Medius Muscle: Comparison of mRNA and Protein Expression Profiles

The major structural protein in skeletal muscle, myosin heavy chain (MyHC), is primarily transcriptionally controlled. We compared the expression of MyHC isoforms on the mRNA and protein level in biopsies from the m. gluteus medius from adult untrained horses. In transverse sections, the majority of fibers showed qualitatively identical mRNA and protein expression patterns. However, coexpression of 2a and 2d/x MyHCs was substantially more common at the protein than at the mRNA level, suggesting a fine-tuning of these two genes in normal muscle not subjected to any training protocol. Because transverse sections give a limited sampling of mRNA expression in the case of uneven distribution of transcripts in a muscle fiber, we also analyzed longitudinal sections. We present, for the first time, evidence that expression of MyHC mRNA and protein was equal along the length of the fiber. Hence, mRNA expression is not regulated by differential expression of isoforms by separate myonuclei. It is concluded that the number of protein hybrid fibers in equine gluteus medius muscle is controlled by alteration of the transcription pattern uniformly along the fiber, rather than by simultaneous transcription of genes. The differences with the results in muscle of small animals and humans are discussed.

Lateral rectus muscle changes after bilateral neonatal labyrinthectomy in the ferret

The vestibular system is essential to the coordination of eye movements during head movements. Exercise, such as the eye movements mediated by the vestibular system, is a major factor in the development of muscle fiber types and the strength of muscle. In this study, the contents of the inner ear were removed (labyrinthectomy) from (LAB) ferrets at postnatal day 10 (P10) and raised with control (SHAM) animals. At P30, the lateral rectus muscles (LR) were removed to analyze the expression of myosin heavy chain (MHC) isoforms and to measure the least diameter of the developmental, slow and fast type muscle fibers. Another set of animals were sacrificed at P120 to analyze MHC isoform expression and muscle fiber diameters, as well as the contractile characteristics of the LR were measured prior to sacrifice. The average LAB LR was significantly stronger than the SHAM LR and the muscle fibers of the LAB animals were larger in diameter. The LAB animals expressed more type IIx and less slow type MHC. These results support the hypothesis that input from the inner ear influence the development of strength and muscle diameter in the ferret extraocular muscles.

Low-intensity exercise training during doxorubicin treatment protects against cardiotoxicity

Doxorubicin (Dox) is a highly effective antineoplastic antibiotic associated with a dose-limiting cardiotoxicity that may result in irreversible cardiomyopathy and heart failure. The purpose of this study was to examine the effects of low-intensity exercise training (LIET) during the course of Dox treatment on cardiac function, myosin heavy chain expression, oxidative stress, and apoptosis activation following treatment. Male Sprague-Dawley rats either remained sedentary or were exercise trained on a motorized treadmill at 15 m/min, 20 min/day, 5 days/wk (Monday through Friday) for 2 wk. During the same 2-wk period, Dox (2.5 mg/kg) or saline was administered intraperitoneally to sedentary and exercised rats 3 days/wk (Monday, Wednesday, Friday) 1-2 h following the exercise training sessions (cumulative Dox dose: 15 mg/kg). Five days following the final injections, hearts were isolated for determination of left ventricular (LV) function, lipid peroxidation, antioxidant enzyme protein expression, 72-kDa heat shock protein expression, caspase-3 activity, and myosin heavy chain isoform expression. Dox treatment significantly impaired LV function and increased caspase-3 activity in sedentary animals (P < 0.05). LIET attenuated the LV dysfunction and apoptotic signal activation induced by Dox treatment and increased glutathione peroxidase expression, but it had no significant effect on lipid peroxidation, protein expression of myosin heavy chain isoforms, 72-kDa heat shock protein, or superoxide dismutase isoforms. In conclusion, our data suggest that LIET applied during chronic Dox treatment protects against cardiac dysfunction following treatment, possibly by enhancing antioxidant defenses and inhibiting apoptosis.

Effect of Endurance Exercise on Myosin Heavy Chain Isoform Expression in Diabetic Rats with Peripheral Neuropathy

This study evaluated the effect of endurance exercise on myosin heavy chain (MHC) isoform expression in soleus muscle of diabetic rats with peripheral neuropathy. Male Sprague Dawley rats were randomly divided into four groups: control sedentary, diabetic sedentary, control exercise, and diabetic exercise. The exercised animals performed treadmill running five times per week. After 12 wks, electrophysiologic testing documented peripheral neuropathy in the diabetic rats. The soleus muscles were then excised and quick-frozen. Cross-sections were immunohistochemically stained for slow, fast, developmental, and neonatal MHCs. Fiber-type composition and fiber cross-sectional areas were then determined. The diabetic groups showed a significantly greater percentage of fast MHC than did the control groups, regardless of exercise status (diabetic sedentary, 22.6%; diabetic exercise, 25.2%; control sedentary, 13.5%; control exercise, 13.1%). The diabetics also showed a significantly lower percentage of slow-only MHC than controls (diabetic sedentary, 77.1%; diabetic exercise, 74.3%; control sedentary, 86.2%; control exercise, 86.1%). No differences in muscle fiber cross-sectional area existed between the groups. The exercised animals showed greater expression of developmental MHC than did the sedentary animals (diabetic sedentary, 1.6%; diabetic exercise, 3.8%; control sedentary, 0.8%; control exercise, 2.0%). The altered slow and fast MHC expression in the diabetic muscle is similar to MHC expression in several other conditions, including decreased neuromuscular activity and denervation. Mechanisms of this MHC expression shift are unknown. Chronic endurance training does not alter adult MHC expression in the diabetic animals. The developmental MHC expression is likely a manifestation of uphill treadmill running due to eccentric contractions in the soleus resulting in myofiber injury and regeneration.

Myosin Heavy Chain Isoform Expression: Influence on Isoinertial and Isometric Performance

Thirty-six healthy men with varying degrees of physical training background performed maximal-effort isometric and isoinertial knee extensor actions, with relative loads equal to 40% and 70% of one-repetition maximum. Force, velocity, and power were derived from force and linear position transducers at 500 Hz. Biopsies were taken from the vastus lateralis and analyzed by SDS-PAGE for relative myosin heavy chain (MHC) content. Relative MHC IIx content was included in a regression model, and explained variance noted. Relative MHC I content was subsequently added to the regression model to determine what, if any, additional variance was explained beyond that of MHC IIx. Results indicated that no relationship (r = 0.0 to 0.1) exists between the relative expression of MHC isoforms from the vastus lateralis and isometric/isoinertial performance in a population with diverse training backgrounds. Lack of nervous system adaptations in the untrained subjects in the study possibly attenuates the significant relationship between MHC and in-vivo muscle performance previously established in trained populations.

Parallel and divergent adaptations of rat soleus and plantaris to chronic exercise and hypergravity

It has been demonstrated that endurance exercise and chronic acceleration, i.e., hypergravity, produce comparable adaptations in a variety of physiological systems, including decreased adiposity, increased energy metabolism, and altered intermediary metabolism. Similar adaptations have not been demonstrated for skeletal muscle per se. To further differentiate between these general responses with respect to gravity and exercise, this study tested the hypothesis that chronic exercise (voluntary wheel running) and chronic acceleration (2 G via centrifugation) will induce similar changes in muscle myosin heavy chain (MHC) isoform expression in rat plantaris, a fast extensor, and in rat soleus, a slow "antigravity" extensor. The experimental design involved four groups of mature male rats (n = 8/group): 1 G and 2 G with running wheels, and 1 G and 2 G controls without running wheels. The primary observations from the study were as follows: 1) 8 wk of 2 G are an adequate stimulus for MHC compositional changes in rat plantaris and soleus muscle; 2) both exercise and +G caused an increase in the slow MHC1 isoform in soleus muscle, suggesting that loading is a primary stimulus for this shift; and 3) 2 G and exercise appeared to have differential effects on the plantaris muscle MHC isoforms, with 2 G causing an increase in MHC2b, and exercise causing a decrease in MHC2b with a concomitant increase in MHC1, suggesting that factors other than enhanced loading, possibly locomotor activity levels, are the primary stimulus for this shift.

Identification of Myosin Heavy Chain I, IIa and IIx in Canine Skeletal Muscles by an Electrophoretic and Immunoblotting Study

To determine which myosin heavy chain (MHC) isoforms are expressed in canine skeletal muscles, different muscle samples of five mixed-breed dogs were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The separated MHC isoforms were identified by immunoblotting technique using a set of specific monoclonal antibodies. To compare the results of the electrophoretic and immunoblotting study, the pattern of MHC isoform expression and histochemical profiles of canine fibres were additionally demonstrated on serial muscle sections by immunohistochemistry and myofibrillar adenosine triphosphatase (mATPase) histochemistry. Not more than three MHC isoforms were demonstrated by SDS-PAGE in the analysed canine muscles. By the immunoblotting technique, the fastest migrating MHC band was identified as slow or MHC-I, the intermediate one as MHC-IIx and the slowest migrating band as MHC-IIa isoform. Since none of the three MHC bands and none of the analysed fibres were recognized by the antibody specific to MHC-IIb of rats, we concluded that MHC-IIb is not expressed in large skeletal muscles of dogs. Similarly, only three major fibre types, i.e. I, IIA and IIX, were revealed according to the pattern of MHC immunohistochemistry and mATPase reaction. Type IIA fibres were more alkali- and acid-stable than type IIX fibres after mATPase histochemistry; hence, the latter corresponded to type IIDog fibres. However, beside the three major fibre types, scarce hybrid fibres co-expressing two MHC isoforms (I/IIA and IIA/IIX) were demonstrated by immunohistochemistry.