Myosin Heavy Chain Isoform Distribution Research Articles

Consequences of Maternal Essential Fatty Acid and Conjugated Linoleic Acid Supplementation on the Development of Calf Muscle and Adipose Tissue.

Simple SummaryCows in modern dairy production systems are fed with silage and concentrate-based diets, resulting in a low essential fatty acid and conjugated linoleic acid status in body tissues and milk. During the late pregnancy and early postnatal period, a reduced maternal fatty acid supply might negatively affect calf development. This study investigated the effects of a maternal re-substitution with essential fatty acids and conjugated linoleic acids on muscle and fat tissue development of neonatal calves. The maternally-supplemented fatty acids were found to be elevated in calf skeletal muscle and fat tissues, but no short-term effects on tissue structure were observed. Nevertheless, the possibility to change offspring fatty acid status via maternal nutrition has been confirmed that could influence offspring development and health in the long-term.Common silage and concentrate-based diets in dairy and beef production may deliver insufficient amounts of essential fatty acids (EFA), thereby also reducing conjugated linoleic acids (CLA) in body tissues and milk. An impaired maternal EFA and CLA supply can have an important impact on calf postnatal development. The current study investigates how maternal supplementation with EFA and CLA affects muscle and adipose tissue development in neonatal calves. Holstein cows (n = 40) were abomasaly supplemented with coconut oil (control), CLA or EFA, or both combined during the transition period. Calves were fed their dam’s colostrum until slaughter at day 5 of life. Fatty acid composition and tissue morphology were analyzed. In muscle and adipose tissues, EFA, CLA, and metabolites were elevated, indicating the effective transfer of maternally-supplemented FA to the offspring. Muscle fiber types, fiber nuclei, myosin heavy chain isoform distribution, capillarization, and fat cell size of intramuscular and other adipose tissues did not differ among groups. The results confirm that maternal nutrition during the transition period can alter the FA composition of the calf tissues. This could influence the offspring’s development and health in the long-term, even though only minor effects were observed in the neonatal calves’ tissue morphology.

Myosin heavy chain isoforms in the myocardium of the atrioventricular junction of Scyliorhinus canicula (Chondrichthyes, Carcharhiniformes).

The atrioventricular junction of the fish heart, namely the segment interposed between the single atrium and the single ventricle, has been studied anatomically and histologically in several chondrichthyan and teleost species. Nonetheless, knowledge about myosin heavy chain (MyHC) in the atrioventricular myocardium remains scarce. The present report is the first one to provide data on the MyHC isoform distribution in the myocardium of the atrioventricular junction in chondrichthyans, specifically in the lesser spotted dogfish, Scyliorhinus canicula, a shark species whose heart reflects the primitive cardiac anatomical design in gnathostomes. Hearts from five dogfish were examined using histochemical and immunohistochemical techniques. The anti-MyHC A4.1025 antibody was used to detect differences in the occurrence of MyHC isoforms in the dogfish, as the fast-twitch isoforms MYH2 and MYH6 have a higher affinity for this antibody than the slow-twitch isoforms MYH7 and MYH7B. The histochemical findings show that myocardium of the atrioventricular junction connects the trabeculated myocardium of the atrium with the trabeculated layer of the ventricular myocardium. The immunohistochemical results indicate that the distribution of MyHC isoforms in the atrioventricular junction is not homogeneous. The atrial portion of the atrioventricular myocardium shows a positive reactivity against the A4.1025 antibody similar to that of the atrial myocardium. In contrast, the ventricular portion of the atrioventricular junction is not labelled, as is the case with the ventricular myocardium. This dual condition suggests that the myocardium of the atrioventricular junction has two contraction patterns: the myocardium of the atrial portion contracts in line with the atrial myocardium, whereas that of the ventricular portion follows the contraction pattern of the ventricular myocardium. Thus, the transition of the contraction wave from the atrium to the ventricle may be established in the atrioventricular segment because of its heterogeneous MyHC isoform distribution. The findings support the hypothesis that a distinct MyHC isoform distribution in the atrioventricular myocardium enables a synchronous contraction of inflow and outflow cardiac segments in vertebrates lacking a specialized cardiac conduction system.

Fetal Sheep Mesenteric Resistance Arteries: Functional and Structural Maturation

Background: Fetal blood pressure increases during late gestation; however, the underlying vascular mechanisms are unclear. Knowledge of the maturation of resistance arteries is important to identify the mechanisms and vulnerable periods for the development of vascular dysfunction in adulthood. Methods: We determined the functional and structural development of fetal sheep mesenteric resistance arteries using wire myography and immunohistochemistry. Results: Media mass and distribution of myosin heavy-chain isoforms showed no changes between 0.7 (100 ± 3 days) and 0.9 (130 ± 3 days) gestation. However, from 0.7 to 0.9 gestation, the resting wall tension increased accompanied by non-receptor-dependent (potassium) and receptor-dependent (noradrenaline; endothelin-1) increases in vasocontraction. Angiotensin II had no contractile effect at both ages. Endothelium-dependent relaxation to acetylcholine and prostaglandin E₂ was absent at 0.7 but present at 0.9 gestation. Augmented vascular responsiveness was paralleled by the maturation of sympathetic and sensory vascular innervation. Non-endothelium-dependent relaxation to nitric oxide showed no maturational changes. The expression of vasoregulator receptors/enzymes did not increase between 0.7 and 0.9 gestation. Conclusion: Vascular maturation during late ovine gestation involves an increase in resting wall tension and the vasoconstrictor and vasodilator capacity of the mesenteric resistance arteries. Absence of structural changes in the tunica media and the lack of an increase in vasoregulator receptor/enzyme expression suggest that vasoactive responses are due to the maturation of intracellular pathways at this gestational age.

Rapid switch-off of the human myosin heavy chain IIX gene after heavy load muscle contractions is sustained for at least four days.

Long-term heavy load contractions decrease the relative amount of the myosin heavy chain (MHC) IIX isoform in human skeletal muscle, but the timing of the down-regulation in the short term is unknown. Untrained subjects performed two resistance bouts, in two consecutive days, with one leg, the other leg serving as a control (age 24±1, n=5). Muscle biopsies were obtained in both legs before, immediately after, and 24, 54, and 96hours after exercise. Serial cryosection analysis combined immunohistochemistry and ATPase histochemistry with In Situ hybridization to identify the distribution of MHC isoforms and their corresponding transcripts, enabling identification of transitional fibers. Fibers positive solely for MHC IIX mRNA decreased in the exercised leg throughout the study period. At 96hours post-exercise, no fibers solely expressed MHC IIX mRNA. In contrast, the number of fibers expressing MHC IIA mRNA increased throughout the study period. The percentage of fibers expressing mRNA for MHC I was unchanged in both legs at all time points. Pronounced depletion of glycogen in the MHC IIX fibers of the exercised leg verifies that the type IIX fibers were active during the heavy load contractions. Major mismatch between MHC at the mRNA and protein levels was only found in the fibers of the exercised leg. These data provide unequivocal in situ evidence of an immediate shutdown of the MHC IIX gene after resistance exercise. A further novel finding was that the silencing of the MHC IIX gene is sustained at least 4days after removal of the stimulus.

Segmental Distribution of Myosin Heavy Chain Isoforms Within Single Muscle Fibers.

Despite many studies looking at the distribution of myosin heavy chain (MHC) isoforms across a transverse section of muscle, knowledge of MHC distribution along the longitudinal axis of a single skeletal muscle fiber has been relatively overlooked. Immunocytochemistry was performed on serial sections of rat extensor digitorum longus (EDL) muscle to identify MHC types I, IIA, IIX, IIY, and IIB. Sixteen fascicles which contained a total of 362 fibers were randomly and systematically sampled from the three EDL muscles. All MHC type I and type II isoforms were expressed. Segmental expression occurred within a very limited segment. MHC isoform expression followed the accepted traditional order from I⇔IIA⇔IIX⇔IIB, however, in some samples expression of an isoform was circumvented from IIB to I or from I to IIB directly. Segmental distribution of MHC isoforms along a single muscle fiber may be because of the myonuclear domain. Anat Rec, 300:1636-1642, 2017. © 2017 Wiley Periodicals, Inc.

Will you still need me (Ca2+, TnT, and DHPR), will you still cleave me (calpain), when I'm 64?

Will you still need me (Ca²⁺, TnT, and DHPR), will you still cleave me (calpain), when I'm 64?

Effects of Resistance Exercise Training on Doxorubicin-Induced Cardiotoxicity.

Although highly effective, doxorubicin (DOX) use is limited by a dose-dependent cardiotoxicity. The purpose of this study was to determine whether resistance training (RT) would protect against DOX-induced cardiac dysfunction and determine whether any observed functional preservation is a result of reduced lipid peroxidation or a preservation of the cardiac myosin heavy chain (MHC) isoform distribution. Rats were resistance-trained or remained sedentary for 12 weeks, then treated with 12.5 mg/kg DOX or 0.9% saline. Five days after DOX exposure, cardiac function, lipid peroxidation, and MHC isoform expression were quantified. RT preserved cardiac function and attenuated the α-to β-MHC shift that occurs with DOX treatment. No significant differences in lipid peroxidation were observed between sedentary and RT animals treated with DOX. These data suggest that resistance-type exercise can provide protection against DOX-induced cardiac dysfunction, which may be a result of a preservation of the cardiac MHC isoform distribution.

Specific protein changes contribute to the differential muscle mass loss during ageing.

In the skeletal muscle, the ageing process is characterized by a loss of muscle mass and strength, coupled with a decline of mitochondrial function and a decrease of satellite cells. This profile is more pronounced in hindlimb than in forelimb muscles, both in humans and in rodents. Utilizing light and electron microscopy, myosin heavy chain isoform distribution, proteomic analysis by 2D-DIGE, MALDI-TOF MS and quantitative immunoblotting, this study analyzes the protein levels and the nuclear localization of specific molecules, which can contribute to a preferential muscle loss. Our results identify the molecular changes in the hindlimb (gastrocnemius) and forelimb (triceps) muscles during ageing in rats (3- and 22-month-old). Specifically, the oxidative metabolism contributes to tissue homeostasis in triceps, whereas respiratory chain disruption and oxidative-stress-induced damage imbalance the homeostasis in gastrocnemius muscle. High levels of dihydrolipoyllysine-residue acetyltransferase (Dlat) and ATP synthase subunit alpha (Atp5a1) are detected in triceps and gastrocnemius, respectively. Interestingly, in triceps, both molecules are increased in the nucleus in aged rats and are associated to an increased protein acetylation and myoglobin availability. Furthermore, autophagy is retained in triceps whereas an enhanced fusion, decrement of mitophagy and of regenerative potential is observed in aged gastrocnemius muscle.

Mechanisms underlying enhancements in muscle force and power output during maximal cycle ergometer exercise induced by chronic β2-adrenergic stimulation in men

The study was a randomized placebo-controlled trial investigating mechanisms by which chronic β2-adrenergic stimulation enhances muscle force and power output during maximal cycle ergometer exercise in young men. Eighteen trained men were assigned to an experimental group [oral terbutaline 5 mg/30 kg body weight (bw) twice daily (TER); n = 9] or a control group [placebo (PLA); n = 9] for a 4-wk intervention. No changes were observed with the intervention in PLA. Isometric muscle force of the quadriceps increased (P ≤ 0.01) by 97 ± 29 N (means ± SE) with the intervention in TER compared with PLA. Peak and mean power output during 30 s of maximal cycling increased (P ≤ 0.01) by 32 ± 8 and 25 ± 9 W, respectively, with the intervention in TER compared with PLA. Maximal oxygen consumption (V̇o2max) and time to fatigue during incremental cycling did not change with the intervention. Lean body mass increased by 1.95 ± 0.8 kg (P ≤ 0.05) with the intervention in TER compared with PLA. Change in single fiber cross-sectional area of myosin heavy chain (MHC) I (1,205 ± 558 μm(2); P ≤ 0.01) and MHC II fibers (1,277 ± 595 μm(2); P ≤ 0.05) of the vastus lateralis muscle was higher for TER than PLA with the intervention, whereas no changes were observed in MHC isoform distribution. Expression of muscle proteins involved in growth, ion handling, lactate production, and clearance increased (P ≤ 0.05) with the intervention in TER compared with PLA, with no change in oxidative enzymes. Our observations suggest that muscle hypertrophy is the primary mechanism underlying enhancements in muscle force and peak power during maximal cycling induced by chronic β2-adrenergic stimulation in humans.

Importance of average litter weight and individual birth weight for early postnatal performance and myofiber characteristics of progeny

Recent evidence suggests that different patterns of prenatal survival causing different levels of intrauterine crowding are occurring in prolific sows bearing litters with 10–15 piglets born, resulting in marked differences in average litter birth weight. Because early intrauterine crowding of embryos is known to have negative effects on prenatal development, the objective of the study was to compare litter characteristics, as well as histomorphometrical and molecular traits of myofibers of selected offspring born from litters of high (H: >1.7kg) and low (L: <1.3kg) average birth weight. The selected newborn littermates were either two females representing the intermediate (HI: 1.73kg) and low (HL: 1.41kg) birth weight from H-litters or two females representing the intermediate (LI: 1.26kg) and high (LH: 1.55kg) birth weight from L-litters. These piglets were sacrificed the day of birth and the longissimus (LM) and semitendinosus (ST) muscles were collected. The average birth weight of the selected HI and LI piglets differed (P<0.05) whereas that of the HL and LH piglets was similar. From birth to weaning, H-offspring grew faster (P<0.05), which was associated with greater (P<0.05) body weight loss of H-compared with L-sows during lactation, where their feed intake did not differ. In LI offspring, brain proportion of birth weight and brain to ST ratio was greater (P<0.05) than in HI but similar between LH and HL progenies. Expressed per mm2, the dark portion of the ST of HI piglets had fewer (P<0.05) primary but similar secondary myofiber numbers compared to LI progeny. Nevertheless, due to the larger ST, total myofiber number was greater in HI and LH than LI piglets. In the LM, myofiber characteristics were similar across birth weight classes, except for the greater (P<0.10) secondary to primary myofiber ratio in HL than LH offspring. In the ST, but not in the LM, the expression of the myoblast differentiation factor 1 was greater (P<0.05) in LH than HL offspring. No birth weight class differences occurred in myosin heavy chain isoform distribution in the LM. In conclusion, female progenies falling in the mid-weight range of the L-litters, but not their heavier littermates, carried some negative phenotypic traits normally associated with intrauterine growth restriction, such as greater brain to ST ratio and impaired myofiber hyperplasia.

Variability in Muscle Adaptation to Electrical Stimulation

The aims were to investigate the plasticity of the myosin heavy chain (MHC) phenotype following neuromuscular electrical stimulation (NMES) and to assess the correlation between MHC isoform distribution and muscle fibre conduction velocity (MFCV).14 men were subjected to 24 sessions of quadriceps NMES. Needle biopsies were taken from the dominant vastus lateralis and neuromuscular tests were performed on the dominant thigh before and after training. NMES significantly increased the quadriceps maximal force by 14.4±19.7% (P=0.02), vastus lateralis thickness by 10.7±8.6% (P=0.01), vastus lateralis MFCV by 11.1±3.5% (P<0.001), vastus medialis MFCV by 8.4±1.8% (P<0.001). The whole spectrum of possible MHC isoform adaptations to training was observed: fast-to-slow transition (4 subjects), bi-directional transformation from MHC-1 and MHC-2X isoforms toward MHC-2A isoform (7 subjects), shift toward MHC-2X (2 subjects), no MHC distribution change (1 subject). No significant correlation was observed between MHC-2 relative content and vastus lateralis MFCV (pre-training: R2=0.04, P=0.46; post-training: R2=0.02, P=0.67). NMES elicited distinct adaptations in the MHC composition and increased force, muscle thickness, and MFCV. The MHC isoform distribution did not correlate with MFCV, thus implying that the proportion of different fibre types cannot be estimated from this electrophysiological variable.

Myosin Isoform Fiber Type and Fiber Size in the Tail of the Virginia Opossum (Didelphis virginiana)

Muscle fiber type is a well studied property in limb muscles, however, much less is understood about myosin heavy chain (MHC) isoform expression in caudal muscles of mammalian tails. Didelphid marsupials are an interesting lineage in this context as all species have prehensile tails, but show a range of tail-function depending on either their arboreal or terrestrial locomotor habits. Differences in prehensility suggest that MHC isoform fiber types may also be different, in that terrestrial opossums may have a large distribution of oxidative fibers for object carrying tasks instead of faster, glycolytic fiber types expected in mammals with long tails. To test this hypothesis, MHC isoform fiber type and their regional distribution (proximal/transitional/distal) were determined in the tail of the Virginia opossum (Didelphis virginiana). Fiber types were determined by a combination of myosin-ATPase histochemistry, immunohistochemistry, and SDS-PAGE. Results indicate a predominance of the fast MHC-2A and -2X isoforms in each region of the tail. The presence of two fast isoforms, in addition to the slow MHC-1 isoform, was confirmed by SDS-PAGE analysis. The overall MHC isoform fiber type distribution for the tail was: 25% MHC-1, 71% MHC-2A/X hybrid, and 4% MHC-1/2A hybrid. Oxidative MHC-2A/X isoform fibers were found to be relatively large in cross-section compared to slow, oxidative MHC-1 and MHC-1/2A hybrid fibers. A large percentage of fast MHC-2A/X hybrids fibers may be suggestive of an evolutionary transition in MHC isoform distribution (fast-to-slow fiber type) in the tail musculature of an opossum with primarily a terrestrial locomotor habit and adaptive tail-function.

Myosin light chain 3f attenuates age‐induced decline in contractile velocity in MHC type II single muscle fibers

Aging is characterized by a progressive loss of muscle mass and impaired contractility (e.g., decline in force, velocity, and power). Although the slowing of contraction speed in aging muscle is well described, the underlying molecular mechanisms responsible for the decrement in speed are unknown. Myosin heavy chain (MHC) isoforms are the primary molecules determining contractile velocity; however, the contraction speed of single fibers within a given MHC isoform type is variable. Recent evidence proposes that the decline in shortening velocity (Vo) with aging is associated with a decrease in the relative content of essential myosin light chain 3f (MLC(3f) ) isoform. In the current study, we first evaluated the relative content of MLC(3f) isoform and Vo in adult and old rats. We then used recombinant adenovirus (rAd) gene transfer technology to increase MLC(3f) protein content in the MHC type II semimembranosus muscle (SM). We hypothesized that (i) aging would decrease the relative MLC(3f) content and Vo in type II fibers, and (ii) increasing the MLC(3f) content would restore the age-induced decline in Vo. We found that there was an age-related decrement in relative MLC(3f) content and Vo in MHC type II fibers. Increasing MLC(3f) content, as indicated by greater % MLC(3f) and MLC(3f) /MLC(2f) ratio, provided significant protection against age-induced decline in Vo without influencing fiber diameter, force generation, MHC isoform distribution, or causing cellular damage. To the best of our knowledge, these are the first data to demonstrate positive effects of MLC(3f) against slowing of contractile function in aged skeletal muscle.

Biochemistry of the Anterior, Medial, and Posterior Genioglossus in the Aged Rat

Age-related tongue weakness may contribute to swallowing deficits in the elderly. One contributing factor may be an alteration in muscle-fiber-type properties with aging. However, it is not clear how muscle fiber types within the aged tongue may vary from those found in young adults, or how fiber types may vary across the anteroposterior axis of the extrinsic tongue muscles. We examined the myosin heavy chain (MHC) composition of anterior, medial, and posterior sections of the genioglossus muscle (GG) in ten old male Fischer 344/Brown Norway rats and compared findings to previously reported data from young adult male rats. Significant differences (p<0.01) between young adult and old rats were found in the distribution of MHC isoforms along the anteroposterior axis of the muscle. In the anterior, medial, and posterior regions, there was a significantly smaller proportion of type IIb MHC in the old rat GG muscles, while the proportion of type IIx MHC was significantly greater. In the medial region, the proportion of type I MHC was found to be significantly greater in the old rats. Thus, we found a shift to more slowly contracting muscle fibers in the aged rat tongue.

Capillary Supply in Relation to Myosin Heavy Chain Fibre Composition of Human Intrinsic Tongue Muscles

The capillary supply and myosin heavy chain (MyHC) composition of three different intrinsic tongue muscles was analysed in the anterior and posterior regions of the human tongue with biochemical and immunohistochemical techniques. Mean capillary density for the whole tongue was 796 ± 82 cap/mm², without regional differences. The overall number of capillaries around each fibre (CAF) was higher in the posterior than in the anterior region (2.5 vs. 2.1, p = 0.009). However, correcting for regional differences in fibre size, CAF per fibre area was higher in the anterior region (4.3 vs. 3.0, p < 0.001). Muscle fibres containing fast MyHCs predominated in the anterior region (78.7%), consisting of MyHCIIa (58.5%), MyHCIIx (1.0%), MyHCIIa+MyHCIIx (11.3%) and MyHCI+MyHCIIa (7.9%). Fibres containing slow MyHC predominated in the posterior region (65.2%), consisting of MyHCI (45.5%) and MyHCI+MyHCIIa (19.7%). A minor fibre population (<2%) contained unusual MyHC isoforms, namely MyHC foetal, MyHC slow-tonic, MyHC α-cardiac or MyHC embryonic. The microvascularization of the human tongue was twice as high as in human limb muscles. Regional similarities in capillary supply, but differences in fibre phenotype composition, suggest that human tongue muscle fibres are fatigue resistant independently of MyHC content. High frequency of hybrid fibres, that is fibres co-expressing two or more MyHC isoforms, indicates a wider spectrum of fibre contractile properties than in limb muscles. In conclusion, human intrinsic tongue muscles showed internal specialization in distribution of MyHC isoforms and capillary supply, but not in the expression of unusual MyHCs.

Long-term leucine supplementation does not increase muscle mass or strength in healthy elderly men

BackgroundIt has been reported that the blunted muscle protein synthetic response to food intake in the elderly can be normalized by increasing the leucine content of a meal. Objective:The objective was to assess the effect of 3 mo of leucine supplementation on muscle mass and strength in healthy elderly men. Design:Thirty healthy elderly men with a mean (±SEM) age of 71 ± 4 y and body mass index (BMI; in kg/m2) of 26.1 ± 0.5 were randomly assigned to either a placebo-supplemented (n = 15) or leucine-supplemented (n = 15) group. Leucine or placebo (2.5 g) was administered with each main meal during a 3-mo intervention period. Whole-body insulin sensitivity, muscle strength (one-repetition maximum), muscle mass (measured by computed tomography and dual-energy X-ray absorptiometry), myosin heavy chain isoform distribution, and plasma amino acid and lipid profiles were assessed before, during, and/or after the intervention period. Results:No changes in skeletal muscle mass or strength were observed over time in either the leucine- or placebo-supplemented group. No improvements in indexes of whole-body insulin sensitivity (oral glucose insulin sensitivity index and the homeostasis model assessment of insulin resistance), blood glycated hemoglobin content, or the plasma lipid profile were observed. Conclusion:Long-term leucine supplementation (7.5 g/d) does not augment skeletal muscle mass or strength and does not improve glycemic control or the blood lipid profile in healthy elderly men. This trial was registered at clinicaltrials.gov as NCT00807508.

Effect of ractopamine-hydrochloride on the fiber type distribution and shelf-life of six muscles of steers

The goal of this study was to evaluate the effects of ractopamine-hydrochloride (RAC) supplementation on the myosin heavy chain isoform distribution and shelf-life properties of muscles from beef top round, knuckle, and loin. Thirty-four steer carcasses were selected from 4 separate slaughter groups. Within each slaughter group (3 groups, n = 8; 1 group, n = 10), steers were separated into pens (n = 8) and fed 0 or 200 mg x animal(-1) x d(-1) of RAC for the final 28 d of feeding. Seventy-two hours postmortem, the longissimus lumborum, semimembranosus (SM), adductor, gracilis, vastus lateralis (VL), and rectus femoris were removed from each carcass. A subsample of each muscle was collected for immunohistochemical analysis. Whole muscles were vacuum packaged and wet aged at 1 +/- 2 degrees C for 13 d before processing into steaks for a 5-d simulated retail display study. Daily, steaks were analyzed for reduction of nitric oxide metmyoglobin, lean color, fat color, and surface discoloration. Objective measures of metmyoglobin, oxymyoglobin, L*, a*, and b* values were recorded daily. Ractopamine significantly (P < 0.05) changed the fiber type isoform distribution in all muscles except the SM. The VL and gracilis presented the greatest fiber type switch with approximately 21% of type I fibers switching to type IIA fibers. However, the fiber type shifts induced by RAC supplementation had little to no effect on subjective and objective color measurements during the 5-d retail display period. Metmyoglobin and oxymyoglobin accumulation, L*, a*, and b*-values were not affected (P > 0.05) by RAC supplementation. Percent nitric oxide metmyoglobin reduction data indicate that reducing ability of RAC-treated steaks from the adductor and longissimus lumborum were significantly affected (P < 0.05). Visual panel data suggest that RAC tended (P < 0.10) to have the most detrimental effect on the lean color and surface discoloration scores of steaks from the VL during the last 3 d of display. Ractopamine significantly (P < 0.05) increased the surface discoloration of the rectus femoris and SM on d 5. Although RAC supplementation had no effect on objective color measurements, subjective measurements indicate that it may have some effect on surface discoloration of some muscles.

Distribution of Myosin Heavy Chain Isoform in Human Extraocular Muscles

Purpose: To provide quantitative data on the distribution of MyHCeom and compare the proportion of myosin heavy chain (MyHC) isoforms between the central and peripheral regions of human extraocular muscles (EOMs). Methods: Medial rectus, lateral rectus, superior rectus, inferior rectus, superior oblique, and inferior oblique muscle samples were taken from three men with brain death. To examine the longitudinal distribution of myosin isoforms, the muscles were divided into central and peripheral portions of equal length. Electrophoresis and densitometry were used to quantify the distribution of MyHC isoforms. Results: Electrophoresis of whole-muscle extracts of sampled EOMs revealed four MyHC bands that were identified as MyHCI, MyHCeom, MyHCIIa, and MyHCIIx. The proportion of MyHCeom was higher in the central region, whereas the proportion of MyHCIIa was higher in the peripheral region. The relative proportions of MyHCI, MyHCeom, and MyHCIIa were not significantly different among the EOMs. There was a tendency for higher levels of MyHCIIx in the inferior rectus muscle. Conclusions: The proportion of MyHCeom was higher in the central region of human EOMs. Further studies are needed to investigate the consequences of this distributional difference on the function of EOMs.

Fiber Type Composition of Cadaveric Human Rotator Cuff Muscles

Descriptive cadaveric laboratory study. To identify the fiber type composition of the rotator cuff and teres major muscles in human subjects. The rotator cuff is commonly injured in athletics and is a major focus of sports medicine. Although the anatomy and architecture of each muscle have been described in great detail, these muscles have never been fiber typed using immunohistochemistry or gel electrophoresis. Fiber typing is important in modeling function, exercise training, and rehabilitation. We harvested tissue samples for all 4 rotator cuff muscles, as well as the teres major muscle from cadavers. Tissues were frozen in liquid nitrogen and sectioned. Cryosections were labeled with commercially available antibodies against fast and slow isoforms of myosin heavy chain (MHC). We also harvested fresh (unembalmed) tissue from deceased subjects and labeled tissue sections with antibodies against fast or slow MHC and wheat germ agglutinin. Gel electrophoresis followed by silver staining was also used to identify and quantify MHC isoforms in fresh tissue samples. All of the muscles were of mixed fiber type composition. As a whole, 44% of rotator cuff fibers labeled positively for slow MHC, with slow MHC content of 54% in supraspinatus, 41% in infraspinatus, 49% in teres minor, 38% in subscapularis, and 40% in teres major. Mixed MHC isoform distribution was confirmed by SDS-PAGE, which also indicated that the IIa and IIx isoforms were roughly equally present across the muscles. Human rotator cuff muscles, at least in older subjects, have a mixed fiber type. Because we only examined older subjects, we must limit our interpretation to this population.

Myosin heavy chain isoform distribution in single fibres of bodybuilders

The purpose of this study was to investigate the long-term effects of high intensity resistance training on myosin heavy chain (MHC) isoform composition of single fibres. Muscle biopsies were obtained from the right vastus lateralis of eight bodybuilders (BB) and seven physical education students (PES). Histochemical analyses were used to determine the fibre type distribution and the fibre cross-sectional area. MHC isoform composition of single fibres was determined with protein electrophoresis. The percentage of fibres expressing MHC IIA and MHC I/IIA was larger in BB (P < 0.05), while MHC IIX was completely absent (P < 0.05). In contrast, myofibrilar ATPase histochemistry only revealed a significantly lower percentage of type IIX fibres in BB (P < 0.05). The muscle fibre profile in the vastus lateralis muscle of BB may represent an adaptation based on the mechanical and biochemical demands of the long-term resistance training.