Myosin Heavy Chain Isoform mRNA Research Articles

Expression of neurotransmitters, vasculogenesis markers and myosin heavy chain isoforms in the masseter muscle of senescence-accelerated mouse prone 8 mice.

Learning and memory deficits and pathologic changes in the hippocampus caused by toothlessness and soft diet feeding are related to reduced masseter muscle (MM) function. Myosin heavy chain (MyHC) isoform expression in the MM also changes under different chewing conditions. The neurotransmitter calcitonin gene-related peptide (CGRP) and vascular endothelial growth factor A (VEGF-A) are involved in MM formation. However, the relationship between CGRP, VEGF-A and MyHC isoforms in the MM in the senescence-accelerated mouse prone 8 (SAMP8) strain, a model of learning and memory deficits, remains unclear. Changes in CGRP, VEGF-A, vasculogenesis marker, and MyHC isoform mRNA expression in the MMs of aging SAMP8 and senescence-accelerated mouse resistant 1 (SAMR1) mice was investigated through quantitative real-time polymerase chain reaction (qRT-PCR) and in situ hybridization. qRT-PCR revealed obviously high CGRP levels in the SAMP8 mouse MM (p<0.001). MyHC-IId/x mRNA expression in the MM was higher in 24-week-old SAMP8 mice than 24-week-old SAMR1 mice (p<0.001) but lower in slow-MyHC SAMP8 mice than SAMR1 mice (p<0.001). CGRP mRNA was observed on the muscle fibers of the SAMP8 mouse MM but not the SAMR1 mouse MM through in situ hybridization. Principal component analysis (PCA) revealed strong positive contributions of SAMP8-MyHC-IId/x, SAMP8-CGRP, SAMR1-MyHC-emb, SAMR1-CGRP, SAMR1-VEGF-A, SAMR1-CD31, SAMP8-VEGF-A, and SAMP8-CD31 in the MM at 12 and 24 weeks. CGRP is also key for the MyHC-IId/x and slow-MyHC patterns in the MMs of SAMP8 mice.

Expression of Myosin Heavy Chain Isoform mRNA Transcripts in the Masseter and Medial Pterygoid Muscles

Expression of Myosin Heavy Chain Isoform mRNA Transcripts in the Masseter and Medial Pterygoid Muscles

Quantification of Myosin Heavy Chain Isoform mRNA Transcripts in the Supraspinatus Muscle of Vertical Clinger Primates

Vertical clinging is a specialized form of locomotion characteristic of the primate family Callitrichidae. Vertical clinging requires these pronograde primates to maintain a vertical posture, so the protraction of their forelimbs must resist gravity. Since pronograde primates usually move as horizontal quadrupeds, we hypothesized that the supraspinatus muscle of vertical clingers would present specific characteristics related to the functional requirements imposed on the shoulder area by vertical clinging. To test this hypothesis, we quantified by real-time quantitative polymerase chain reaction the mRNA transcripts of myosin heavy chain (MHC) isoforms in the supraspinatus muscle of 15 species of pronograde primates, including vertical clingers. Our results indicate that the supraspinatus of vertical clingers has a specific expression pattern of the MHC isoforms, with a low expression of the transcripts of the slow MHC-I isoform and a high expression of the transcripts of the fast MHC-II isoforms. We conclude that these differences can be related to the particular functional characteristics of the shoulder in vertical clingers, but also to other anatomical adaptations of these primates, such as their small body size.

Acute Myosin Heavy Chain Isoform mRNA Expression in Response to Two Resistance Exercise Intensities With Equal Volume Load in Resistance-Trained Men.

The purpose of this study was to determine if resistance exercise intensity, in the context of equal volume load, differentially affected myosin heavy chain (MHC) isoform messenger RNA (mRNA) expression in resistance-trained men. In a crossover, uniform-balanced design, 10 male participants (23.7 ± 2.8 years, 178.8 ± 5.9 cm, 85.9 ± 9.2 kg) completed 2 lower-body resistance exercise sessions of different intensities with equal volume load. For the higher-intensity exercise session, participants performed 5 sets of 6 repetitions at 80% of 1 repetition maximum (1RM). For the lower-intensity exercise session, participants performed 3 sets of 16 repetitions at 50% of 1RM. Muscle samples from the vastus lateralis were acquired before exercise (PRE), 45 minutes postexercise (45MINPE), 3 hours postexercise (3HRPE), 24 hours postexercise (24HRPE), and 48 hours postexercise (48HRPE). Statistical analyses of mRNA expression were performed using separate 2 × 5 two-way repeated-measures analyses of variance for each criterion variable (p ≤ 0.05). There were no statistically significant interactions between intensity and time. Likewise, there were no significant differences between exercise intensity in MHC expression. Expression of mRNA for all MHC isoforms decreased at all postexercise time points, except 3HRPE (p = 0.051), compared with PRE following both exercise bouts (p ≤ 0.05). The results of this study found no difference in mRNA expression of MHC isoforms as a function of resistance exercise intensity. In addition, in contrast to results found in previous studies of untrained men, MHC mRNA expression seems to decrease in response to acute resistance exercise in previously resistance-trained men.

Porcine satellite cells are restricted to a phenotype resembling their muscle origin

Muscles in most domestic animals differ in function and growth potential based largely on muscle fiber type composition. Though much is known about satellite cells (SC), information is limited regarding how populations of SC differ with muscle fiber type, especially in pigs. Therefore, the objective of this study was to isolate and culture SC from red (RST) and white (WST) portions of the semitendinosus muscle of neonatal and adult pigs and determine their capacity to proliferate, differentiate, and express various myosin heavy chain (MyHC) isoforms in vitro. Porcine satellite cells were isolated from RST and WST muscles of 6-wk-old and adult (>6-mo-old) pigs and cultured under standard conditions. Muscle from neonatal pigs yielded nearly 10 times more (P < 0.001) presumptive satellite cells as those from adult pigs, with fusion percentages close to 60% for the former. The RST yielded more (P < 0.001) SC per gram muscle compared to WST, 8.1 ± 0.2 × 10(4) cells versus 6.7 ± 0.1 × 10(4) cells/gram muscle in young pigs, and 9.7 ± 0.4 × 10(3) cells versus 5.5 ± 0.4 × 10(3) cells/gram muscle in adult pigs, respectively. Likewise, satellite cells from RST proliferated faster (P < 0.001) than those from WST across both ages, as indicated by a shorter cell doubling time, 18.6 ± 0.8 h versus 21.3 ± 0.9 h in young pigs, and 23.2 ± 0.7 h versus 26.7 ± 0.9 h in adult pigs, respectively. As a result of shorter times to confluence, satellite cells from RST also formed myotubes earlier than those SC originating from WST. Once induced, however, SC from WST differentiated and fused faster (P < 0.05) as evidenced by fusion percentage within the first 24 h, 41.6% versus 34.3%, respectively; but reached similar ultimate fusion percentages similar to WST by 48 h. Over 90% of MyHC expressed in maximally fused SC cultures from both RST and WST was restricted to the embryonic isoform. Type IIX MyHC mRNA was not detected in any culture. Myotube cultures from RST expressed more (P < 0.01) Type I MyHC isoform mRNA than those from WST, whereas those cultures from WST expressed more (P < 0.05) Type II (including Types IIA and IIB) MyHC transcripts. These data show SC cultures from porcine fast and slow muscles express MyHC profiles largely reflective of their muscle of origin and suggest satellite cells are partially restricted to a particular muscle phenotype in which they are juxtapositioned. Understanding the molecular nature of these intrinsic control mechanisms may lead to improved strategies for augmenting meat animal growth or muscle regeneration.

Administration of estradiol, trenbolone acetate, and trenbolone acetate/estradiol implants alters adipogenic and myogenic gene expression in bovine skeletal muscle1

Twenty crossbred yearling steers (421 kg) were used to evaluate the effects of implanting with trenbolone acetate (TBA; 120 mg), estradiol-17β (E(2); 25.7 mg), and a combination (120 mg of TBA and 24 mg of E(2)) on adipogenic and myogenic mRNA concentrations. Animals were blocked by BW and within each block were assigned to 1 of 4 treatments. Animals were housed and fed in individual pens with 5 animals per treatment. All animals were weighed weekly, and muscle biopsy samples were taken from the LM of each steer on d 0 (before implantation), 7, 14, and 28. Total RNA was isolated from each sample and real-time quantitative PCR was used to measure the quantity of C/EBPβ, PPARγ, stearoyl CoA desaturase (SCD), myogenin, and 3 isoforms of bovine myosin heavy chain (MHC) mRNA. Total BW gain from the 28-d period was adjusted to d 0 by use of covariant analysis, and steers in the implant groups tended (P = 0.09) to have increased BW gain compared with nonimplanted control steers. Analysis of the gene expression of MHC showed that neither implant nor day (P > 0.20) had a significant effect on the expression of type I or IIX MHC mRNA There was also no treatment effect (P > 0.20) on MHC-IIA and myogenin, but increasing days on feed increased (P = 0.05) the expression of MHC-IIA mRNA. Relative mRNA abundance of C/EBPβ, PPARγ, and SCD increased (P < 0.05) during days of feed but PPARγ decreased (P < 0.05) with the treatment of combined TBA/E(2) implant. Results of this study indicate that implanting with TBA, E(2), or both increased BW gain and decreased adipogenic gene expression of finishing steers without significantly affecting the concentration of type I, IIA, or IIX MHC mRNA. Increasing days on feed increased both MHC-IIA and adipogenic gene expression in bovine skeletal muscle biopsy samples. We conclude that administration of steroidal implants had no effect on the proportion of the 3 MHC mRNA isoforms but decreased C/EBPβ, PPARγ, and SCD mRNA in bovine skeletal muscle.

Administration of estradiol, trenbolone acetate, and trenbolone acetate/estradiol implants alters adipogenic and myogenic gene expression in bovine skeletal muscle

Twenty crossbred yearling steers (421 kg) were used to evaluate the effects of implanting with trenbolone acetate (TBA; 120 mg), estradiol-17β (E(2); 25.7 mg), and a combination (120 mg of TBA and 24 mg of E(2)) on adipogenic and myogenic mRNA concentrations. Animals were blocked by BW and within each block were assigned to 1 of 4 treatments. Animals were housed and fed in individual pens with 5 animals per treatment. All animals were weighed weekly, and muscle biopsy samples were taken from the LM of each steer on d 0 (before implantation), 7, 14, and 28. Total RNA was isolated from each sample and real-time quantitative PCR was used to measure the quantity of C/EBPβ, PPARγ, stearoyl CoA desaturase (SCD), myogenin, and 3 isoforms of bovine myosin heavy chain (MHC) mRNA. Total BW gain from the 28-d period was adjusted to d 0 by use of covariant analysis, and steers in the implant groups tended (P = 0.09) to have increased BW gain compared with nonimplanted control steers. Analysis of the gene expression of MHC showed that neither implant nor day (P > 0.20) had a significant effect on the expression of type I or IIX MHC mRNA There was also no treatment effect (P > 0.20) on MHC-IIA and myogenin, but increasing days on feed increased (P = 0.05) the expression of MHC-IIA mRNA. Relative mRNA abundance of C/EBPβ, PPARγ, and SCD increased (P < 0.05) during days of feed but PPARγ decreased (P < 0.05) with the treatment of combined TBA/E(2) implant. Results of this study indicate that implanting with TBA, E(2), or both increased BW gain and decreased adipogenic gene expression of finishing steers without significantly affecting the concentration of type I, IIA, or IIX MHC mRNA. Increasing days on feed increased both MHC-IIA and adipogenic gene expression in bovine skeletal muscle biopsy samples. We conclude that administration of steroidal implants had no effect on the proportion of the 3 MHC mRNA isoforms but decreased C/EBPβ, PPARγ, and SCD mRNA in bovine skeletal muscle.

Association between the lack of teeth and the expression of myosins in masticatory muscles of microphthalmic mouse

The purposes of the present study were to elucidate the influences of the deficiency of teeth on masticatory muscles, such as the masseter, temporalis and digastric muscles and compare the influence among masticatory muscles. We analysed the expressions of myosin heavy chain (MyHC) isoform messenger RNA (mRNA) and protein in these muscles in the microphthalmic (mi/mi) mouse, whose teeth cannot erupt because of a mutation in the mitf gene locus. The expression levels of MyHC mRNA and protein in the masseter, temporalis, digastric, tibialis anterior and gastrocnemius muscles of +/+ and mi/mi mice were analysed with real-time polymerase chain reaction and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. The mi/mi masseter muscle at 8 weeks of age expressed 4.1-fold (p < 0.05) and 3.3-fold (p < 0.01) more MyHC neonatal mRNA and protein than that in the +/+, respectively; the expression level of MyHC neonatal protein was 19% of the total MyHC protein in the masseter muscle of mi/mi mice. In the digastric muscle, the expression levels of MyHC I mRNA and protein in the mi/mi mice were 4.7-fold (p < 0.05) and 5-fold (p < 0.01) higher than those in the +/+ mice. In the temporalis, tibialis anterior and gastrocnemius muscles, there was no significant difference in the expression levels of any MyHC isoform mRNA and protein between +/+ and mi/mi mice. These results indicate associations between the lack of teeth and the expression of MyHC in the masseter and digastric muscles but not such associations in the temporalis muscle, suggesting that the influence of tooth deficiency varies among the masticatory muscles.

Myosin heavy chain mRNA isoforms are expressed in two distinct cohorts during C2C12 myogenesis

The regulation of muscle fibre transitions has mainly been studied in vivo using conventional histological or immunohistochemical techniques. In order to investigate the molecular regulation of myosin heavy chain (MyHC) isoform expression in cell culture studies, we first characterised the normal transitions in endogenous expression of the MyHC isoforms and the myogenic regulatory factors during differentiation of C2C12 muscle cells. Interestingly, across the time course of differentiation, MyHC mRNA isoforms were expressed in a distinct temporal pattern as two distinct cohorts, one including MyHC I, embryonic and neonatal, the other including MyHC IIa, IIx and IIb. The pattern of expression suggests a transition in MyHC isoforms, from one cohort to another, occurs during muscle cell differentiation and that these transitions occur independent of nerve innervation. To our knowledge, this is the most comprehensive analysis of in vitro MyHC mRNA isoform transitions and provides important information for studying the regulation of transitions in MyHC isoforms in cell culture systems.

Loss of IGF-IEa or IGF-IEb Impairs Myogenic Differentiation

Actions of protein products resulting from alternative splicing of the Igf1 gene have received increasing attention in recent years. However, the significance and functional relevance of these observations remain poorly defined. To address functions of IGF-I splice variants, we examined the impact of loss of IGF-IEa and IGF-IEb on the proliferation and differentiation of cultured mouse myoblasts. RNA interference-mediated reductions in total IGF-I, IGF-IEa alone, or IGF-IEb alone had no effect on cell viability in growth medium. However, cells deficient in total IGF-I or IGF-IEa alone proliferated significantly slower than control cells or cells deficient in IGF-IEb in serum-free media. Simultaneous loss of both or specific loss of either splice variant significantly inhibited myosin heavy chain (MyHC) immunoreactivity by 70-80% (P < 0.01) under differentiation conditions (48 h in 2% horse serum) as determined by Western immunoblotting. This loss in protein was associated with reduced MyHC isoform mRNAs, because reductions in total IGF-I or IGF-IEa mRNA significantly reduced MyHC mRNAs by approximately 50-75% (P < 0.05). Loss of IGF-IEb also reduced MyHC isoform mRNA significantly, with the exception of Myh7, but to a lesser degree (∼20-40%, P < 0.05). Provision of mature IGF-I, but not synthetic E peptides, restored Myh3 expression to control levels in cells deficient in IGF-IEa or IGF-IEb. Collectively, these data suggest that IGF-I splice variants may regulate myoblast differentiation through the actions of mature IGF-I and not the E peptides.

Chronic activation of 5'-AMP-activated protein kinase changes myosin heavy chain expression in growing pigs

The purpose of this study was to determine the effect of 5'-AMP-activated protein kinase (AMPK) on energy metabolism and myosin heavy chain (MyHC) isoform expression in growing pigs using chronic treatment with 5-aminoimidazole-4-carboxamide-1-beta-d-ribofuranoside (AICAR) as a model. Four-week-old pigs were given daily injections of AICAR or 0.9% saline for 10 d. Treatment with AICAR increased (P < 0.05) AMPK activity in semitendinosus muscles (STM). Expression of skeletal muscle specific glucose transporter 4 (GLUT4) was also enhanced (P < 0.05) by AICAR treatment. Using real-time PCR, electrophoresis, and Western blot analyses, we confirmed that AICAR treatment caused a decrease (P < 0.05) in type IIa MyHC isoform mRNA and protein levels and a concomitant increase (P < 0.05) in type IIx MyHC containing fibers. Consistent with a MyHC isoform shift from IIa to IIx, muscles from pigs treated with AICAR had greater (P < 0.05) lactate dehydrogenase (LDH) activity. Moreover, muscle of treated pigs expressed greater (P < 0.05) message for LDH. Administration of AICAR, however, did not alter expression of PPAR-gamma coactivator-1alpha, fatty acid translocase, citrate synthase, or the activity of cytochrome c oxidase. Overall, these results indicate that activation of AMPK by AICAR causes muscle to assume a faster-contracting, more glycolytic nature. These data are in direct contrast to documented effects in rodent models, but these effects may be dependent on the time of administration and the overall growth status of the animal.

Novel epigenetic regulation of skeletal muscle myosin heavy chain genes. Focus on “Differential epigenetic modifications of histones at the myosin heavy chain genes in fast and slow skeletal muscle fibers and in response to muscle unloading”

skeletal muscle is a highly plastic tissue, capable of responding and adapting to a variety of physiological stimuli, e.g., muscle loading and unloading. Multiple functional phenotypes of skeletal muscle exist throughout the body at the same time. For example, muscles required for antigravity support of the skeleton or sustained locomotion are generally slow-contracting, low force-generating, and relatively resistant to fatigue, while other muscles required for quick, powerful movements are generally fast-contracting, high force-generating, and easily fatigued. In fact, skeletal muscle phenotypes have been characterized using combinations of muscle properties, including metabolic, fatigability, color, innervation, or predominant contractile protein expression. Myosin heavy chain (MHC) is the most abundant protein in skeletal muscle, comprising ∼25% of the total protein content, and is a major component of the complex responsible for generating contractile force in skeletal muscle. At least nine MHC isoforms, each transcribed from their own gene, exist in mammalian striated muscle; but only slow type I MHC and various isoforms of fast type II (IIa, IIx/d, and IIb MHC isoforms) are present in adult limb skeletal muscle that is not undergoing regeneration (16). It is well established that skeletal muscle phenotype, particularly MHC gene expression, can be altered by different states of muscular activity or inactivity, as well as various hormonal and metabolic factors (1).

Mechanical overload increases muscle mass but does not change myosin isoform expression in a rodent hibernator, Spermophilus lateralis

Hibernating ground squirrels do not show slow to fast myosin heavy‐chain (MyHC) isoform transitions with atrophy, and we test whether increased muscle loading would also fail to produce typical MyHC isoform shifts. Field‐caught, Summer‐active ground squirrels (Spermophilus lateralis) were assessed for the responsiveness of hindlimb plantaris muscles to mechanical overload through synergist surgical ablation. Plantaris muscles of surgical animals were 40% larger than controls after 14 days, but did not show any of the expected shifts in MyHC isoform expression. We also measured MyHC isoform mRNA expression, citrate synthase activity, and the expression of a suite of genes involved in the control of skeletal muscle mass and MyHC isoform expression, including MAFbx, MuRF1, FOXO1, IGF1 and IGFBP5, and myostatin. Our results show an unusual insensitivity of hibernator muscles to mechanical overload induced isoform shifts, yet muscle mass was increased, which has implications for isoform expression and muscle adaptations during hibernation. Funding sources: NIH SCORE, NIH RISE, CSULB.

Myosin heavy chain mRNA isoforms in masseter muscle before and after orthognathic surgery

Orthognathic surgery leads to changed jaw position and force vector of mastication to which the muscles must adapt. The aim of the present study was to determine the relative expression of myosin heavy chain (MyHC) messenger RNA (mRNA) isoforms in different types of human masseter muscle fiber under consideration of change in the number of occlusal contacts before and 6 months after surgery. Muscle biopsies were taken from the anterior and posterior parts of both sides in 30 patients with prognathic and retrognathic mandibles. Specific mRNA MyHC analysis was made by real-time polymerase chain reaction to quantify the isoforms I, IIa, and IId/x. There was a shift in the relative content from type I (46% before, 37% after) to type IIa (29% before, 42% after). This shift correlates with number of teeth in occlusion. Correlation between isoform shift and number of teeth in occlusion indicates higher mastication force which stabilizes the treatment result.

Effects of Varying Intensity Resistance Exercise on Myogenic Regulatory Factors and Myosin Heavy Chain Genes

A single bout of resistance training is capable of activating the expression of many diverse groups of genes including myosin heavy chain (MHC) isoforms and the myogenic regulatory factors (MRF). However, the role of exercise intensity in regards to the effects on transcription and protein synthesis are not well denned. PURPOSE: The purpose of this study was to investigate the effects of resistance exercise intensity on MRF mRNA and protein and MHC mRNA isoforms. METHODS: Thirteen male participants (21.5 ± 2.9 yrs, 86.1 ± 19.5 kg, 69.7 ± 2.7in) completed two bouts [low-intensity/high volume (LI = 65% 1-RM) and high-intensity/ low volume (HI = 85% 1-RM)] of single-legged resistance exercise, each employing four sets of leg press and leg extension. Muscle biopsies were obtained from each leg immediately before exercise (PRE), and at 30 minutes (30MPST), 2 hours (2HRPST), and 6 hours (6HRPST) following each resistance exercise bout. Data were analyzed using RT-PCR for mRNA expression and ELISA for protein expression. Statistical analyses were performed by utilizing a 2 x 4 MANOVA (p ≤ 0.05). RESULTS: Results indicated that MHC I, IIb, and IIx all significantly increased in response to resistance exercise (p < 0.001). Furthermore, exercise intensity played a role with LI exercise stimulating a more positive response in MHC I (p < 0.001) and HI eliciting a greater response in MHC IIb and IIx (p < 0.001). In regards to the MRF, resistance exercise significantly increased the expression of Myogenin, Myf5, and MRF-4 mRNA (p < 0.05). Furthermore, LI exercise stimulated a more positive response in MyoD, Myogenin, and Myf5 mRNA (p < 0.05), as well as MyoD, Myogenin, and MRF-4 protein (p < 0.05). MRF-4 protein also significantly increased at 6HRPST over the baseline values (p < 0.05). Changes in MyoD and Myogenin mRNA were also found to be significantly correlated (p < 0.05) to changes in MHC I, IIa, IIb, and IIx mRNA. CONCLUSIONS: This study suggests that both MHC and MRF mRNA expression is quickly up-regulated in response to resistance exercise, but that LI and high volume may generate a more robust response.

Highlights From The Literature

Highlights From The Literature

Mechanisms underlying myosin heavy chain expression during development of the rat diaphragm muscle

During early postnatal development in rat diaphragm muscle (Dia(m)), significant transitions in myosin heavy chain (MHC) isoform expression occur that are associated with fiber growth and increased MHC protein. At present, there is no direct information regarding the transcriptional regulation of MHC isoform expression during postnatal Dia(m) development. We hypothesized postnatal changes in MHC isoform mRNA expression are followed by concomitant changes in MHC protein expression. The Dia(m) was removed at postnatal days 0, 14, 28, and 84 (adult). MHC mRNA expression was determined by real-time RT-PCR. MHC protein expression was determined by SDS-PAGE. There was a significant effect of postnatal age on MHC isoform mRNA and protein expression. At birth, the MHC(Neo) isoform accounted for 28% of MHC mRNA and 54% of total MHC protein. By postnatal day 14, MHC(Neo) mRNA and protein increased significantly, and both decreased significantly by day 28, consistent with transcriptional control of the expression of this developmental isoform. By postnatal day 28, there were minimal changes in mRNA expression for MHC(Slow) and MHC(2X), yet protein expression increased significantly. MHC(2A) mRNA and protein expression did not change during this time. Thus changes in MHC protein expression did not follow (or parallel) changes in MHC mRNA for the adult MHC isoforms. The present findings indicate that changes in MHC expression in the developing rat Dia(m) are not driven solely by changes in mRNA expression. Knowledge of isoform-specific MHC mRNA expression only yields predictive information on MHC protein expression for the MHC(Neo) isoform.

The effect of magnetic stimulation on unloaded soleus muscle of rat: changes in myosin heavy chain mRNA isoforms

This study assessed the potential application and the effectiveness of functional magnetic stimulation (FMS) for preventing skeletal muscle atrophy in adult rats. FMS using magnetic stimulator was performed to rat soleus muscle by placing a round magnetic coil on the back of 3rd-5th lumbar vertebral level at 20 Hz frequency for 60 min/day up to 10 days. A reverse transcriptase-polymerase chain reaction was applied to evaluate relative amounts of mRNAs specific to four myosin heavy chain (MHC) isoforms [MHCIbeta, MHCIIa, MHCIIb, and MHCIId(x)] in rat soleus muscle during contractile activity by magnetic stimulation. Ten-day unloading by hindlimb suspension induced a drastic decrease of MHCIbeta and MHCIIa mRNA expressions, while MHCIIb and MHCIId(x) mRNA was not decreased. The magnetic stimulation resuscitated the down-regulation of the mRNA levels of MHCIbeta and MHCIIa. These results suggest that magnetic stimulation on acute atrophied muscles is useful for preventing the muscle atrophy.

Myosin heavy chain isoform mRNA and protein levels after long-term paralysis

To assess the long-term influence of paralysis on muscle phenotypic mRNA and protein expression, the effects of spinal cord transection (ST) on myosin heavy chain (MyHC) isoform mRNA and protein levels in the soleus and medial gastrocnemius (MG) muscles of rats were analyzed. Control soleus contained predominantly MyHC-I with low amounts of MyHC-IIa and IIx mRNAs. After ST, MyHC-I mRNA decreased to ∼15%, MyHC-IIa was increased by 75–200%, and MyHC-IIx was elevated by 8–10×. Low level expression of MyHC-IIb was observed post-ST, suggesting that reduced activity is not a primary stimulus for MyHC-IIb expression. Adaptations in mRNA preceded protein adaptations in the soleus. Although MyHC-I protein in the MG was reduced post-ST, no other consistent changes occurred. The relative lack of adaptation to ST by the MG suggests that the reduced activity and load bearing encountered by the MG were insufficient to induce a change in muscle phenotype.

Age and Exercise Effects on Myosin Heavy Chain Isoform mRNA Expression in Human Muscle

0389 Aging is associated with changes in the contractile and metabolic properties of human skeletal muscle. Among these changes is a shift in fiber type towards a predominance of Type I fibers. It was reported that mRNA levels of myosin heavy chain (MHC) isoforms, the determinant of contractile phenotype, are altered with age in a manner consistent with fiber type results, although two studies found no changes in MHC mRNA expression. Further, a predominance of Type I fibers has been observed in highly aerobically-trained athletes but the effect of aerobic training on MHC mRNA expression and the potential interaction with age remains to be determined. PURPOSE: To determine the effect of aging and a moderate intensity aerobic exercise training program on mRNA levels of MHC isoforms in human muscle. METHODS: Previously sedentary, healthy men and women aged 22–88 y were studied before and after 16 weeks of bicycle training (30–45 min at 70–80% peak heart rate, 3–4 days/week, N = 62) or control (stretching, N = 37) activity. Muscle biopsies of the vastus lateralis were analyzed for MHC mRNA isoforms I, IIA and IIX using real-time PCR. RESULTS: At baseline MHC-I mRNA was unchanged with age, while levels of IIA (r = −0.45) and IIX (r = −0.34) declined by 14% and 10% per decade, respectively (p < 0.001). After training, the mRNA levels of MHC-I and IIA increased 61 and 99%, respectively, while IIX decreased 50% (all p < 0.001). The increase in MHC-I mRNA was positively associated with age (r = 0.30, p < 0.01). There was no interaction between age and exercise responses for MHC-IIA or IIX, and MHC mRNA did not change in the control group. CONCLUSION: This study demonstrates that MHC isoform expression changes with aging consistent with reported changes in fiber type. Despite baseline differences, older muscle retains the ability to alter expression of MHC isoforms at the mRNA level in response to aerobic training. This agrees with previous data from this study demonstrating that training-induced enhancement of mitochondrial genes and functions are similar among younger and older people. Thus, skeletal muscle plasticity, at least at the transcript level, remains intact in older people. Future work will determine if training-induced changes in MHC mRNA are also reflected at the protein level. Supported by NIH AG-RO109531, MO1-RR00585 and Mayo Foundation.