Changes In Muscle Fiber Types Research Articles

Novel Candidate Genes Involved in an Initial Stage of White Striping Development in Broiler Chickens.

White striping (WS) is a myopathy characterized by the appearance of white stripes parallel to the muscle fibers in the breast of broiler chickens, composed of adipose and connective tissues. This condition causes economic losses and, although common, its etiology remains poorly understood. Hence, the objective was to identify genes and biological mechanisms involved in the early stages of WS using a paternal broiler line that grows slightly slower than commercial ones, at 35 days of age, through the RNA sequencing of the pectoralis major muscle. Thirty genes were differentially expressed between normal and WS-affected chickens, with 23 upregulated and 7 downregulated in the affected broilers. Of these, 14 genes are novel candidates for WS and are implicated in biological processes related to muscle development (CEPBD, DUSP8, METTL21EP, NELL2, and UBE3D), lipid metabolism (PDK4, DDIT4, FKBP5, DGAT2, LIPG, TDH, and RGCC), and collagen (COL4A5 and COL4A6). Genes related to changes in muscle fiber type and the processes of apoptosis, autophagy, proliferation, and differentiation are possibly involved with the initial stage of WS development. In contrast, the genes linked to lipid metabolism and collagen may have their expression altered due to the progression of the myopathy.

Skeletal muscle adaptations to high-intensity, low-volume concurrent resistance and interval training in recreationally active men and women.

This study compared the structural and cellular skeletal muscle factors underpinning adaptations in maximal strength, power, aerobic capacity, and lean body mass to a 12-week concurrent resistance and interval training program in men and women. Recreationally active women and men completed three training sessions per week consisting of high-intensity, low-volume resistance training followed by interval training performed using a variety upper and lower body exercises representative of military occupational tasks. Pre- and post-training vastus lateralis muscle biopsies were analyzed for changes in muscle fiber type, cross-sectional area, capillarization, and mitochondrial biogenesis marker content. Changes in maximal strength, aerobic capacity, and lean body mass (LBM) were also assessed. Training elicited hypertrophy of type I (12.9%; p = 0.016) and type IIa (12.7%; p = 0.007) muscle fibers in men only. In both sexes, training decreased type IIx fiber expression (1.9%; p = 0.046) and increased total PGC-1α (29.7%, p < 0.001) and citrate synthase (11.0%; p < 0.014) content, but had no effect on COX IV content or muscle capillarization. In both sexes, training increased maximal strength and LBM but not aerobic capacity. The concurrent training program was effective at increasing strength and LBM but not at improving aerobic capacity or skeletal muscle adaptations underpinning aerobic performance.

Human myofiber-enriched aging-induced lncRNA FRAIL1 promotes loss of skeletal muscle function.

The loss of skeletal muscle mass during aging is a significant health concern linked to adverse outcomes in older individuals. Understanding the molecular basis of age-related muscle loss is crucial for developing strategies to combat this debilitating condition. Long noncoding RNAs (lncRNAs) are a largely uncharacterized class of biomolecules that have been implicated in cellular homeostasis and dysfunction across a many tissues and cell types. To identify lncRNAs that might contribute to skeletal muscle aging, we screened for lncRNAs whose expression was altered in vastus lateralis muscle from older compared to young adults. We identified FRAIL1 as an aging-induced lncRNA with high abundance in human skeletal muscle. In healthy young and older adults, skeletal muscle FRAIL1 was increased with age in conjunction with lower muscle function. Forced expression of FRAIL1 in mouse tibialis anterior muscle elicits a dose-dependent reduction in skeletal muscle fiber size that is independent of changes in muscle fiber type. Furthermore, this reduction in muscle size is dependent on an intact region of FRAIL1 that is highly conserved across non-human primates. Unbiased transcriptional and proteomic profiling of the effects of FRAIL1 expression in mouse skeletal muscle revealed widespread changes in mRNA and protein abundance that recapitulate age-related changes in pathways and processes that are known to be altered in aging skeletal muscle. Taken together, these findings shed light on the intricate molecular mechanisms underlying skeletal muscle aging and implicate FRAIL1 in age-related skeletal muscle phenotypes.

Comparison of muscle fiber characteristics and meat quality between newborn and adult Haimen goats

Goat meat is popular with consumers for its rich nutritional content. Muscle fiber characteristics have been shown to play a crucial role in determining the quantity and quality of meat. However, little is known about the temporal changes in muscle fiber characteristics and meat quality during growth in goats. In this study, muscle fiber type, fiber diameter, fiber cross-sectional area (CSA), glycolytic potential (GP), meat pH, and meat color were analyzed in the gastrocnemius (GAS), gluteus medius (GM), biceps brachii (BB), longissimus lumborum (LL) muscles from newborn (NHMG) and adult (AHMG) Haimen goats. The distribution of type I and type Π fiber in goats is not consistent across the four muscles and undergoes alterations with age. The diameter and CSA of the muscle fibers were similar among the four NHMG muscles. However, in AHMG, the LL muscle had the largest fiber in terms of both diameter and CSA, followed by BB, GM, and GAS muscles. Moreover, the CSA of type Π fibers was higher than that of type I fibers in both NHMG and AHMG. GP values ranged from 90 to 140 umol/g across the muscle and no significant differences were observed. AHMG had a higher pH level and a* value, but lower L* and b* values than NHMG. Overall, our findings enhance our understanding of the changes in muscle fiber type and meat quality during the growth in Haimen goats and provide a basis for future research on the development and transformation of muscle fibers in goats.

THU658 My Giant Calves Gave Up My Diagnosis: An Extreme Manifestation Of A Very Rare Syndrome With A Severe Elevation Of Tsh Levels

Abstract Disclosure: E.E. Castro Feliu: None. A. Ludena De Freitas: None. A. Torres Rodríguez: None. G. Irizarry: None. Hoffmann’s Syndrome is a rare type of hypothyroidism myopathy in adults that classically presents with muscle cramps, weakness, muscle stiffness, hyporeflexia, delay deep tendon reflexes and pseudohypertrophy. This is the case of a 55-year-old male with history of poorly controlled hypothyroidism and medication non-compliance who was admitted to our institution for non-purulent cellulitis. Vital signs were within normal limits and Body Mass Index was 41.1. Upon physical examination flat affect, periorbital edema, moon facies features and macroglossia were distinguished. Skin had a doughy texture with areas of ichthyosis at upper and lower extremities. Goiter was absent. The most remarkable finding was bilateral calf hypertrophy, with measurements of 56 cm and 55.5 cm in diameter of right and left calves respectively. There was no tenderness to calf palpation and Homan’s sign was absent. Laboratories revealed dyslipidemia with a cholesterol of 308 mg/dL, triglycerides of 782 mg/dL, HDL of 26 mg/dL and LDL of 216 mg/dL with elevated inflammatory markers including sedimentation rate of 77 mm/hr and C-reactive protein of 6.45 mg/dL. Hypothyroidism was suspected therefore thyroid studies were ordered, which was compatible with severe hypothyroidism demonstrated by a record level TSH of 523 lU/ml. Other reported levels were Free T3 0.94pg/ml, total T3 0.5 ng/ml, free thyroxine &amp;lt;0.07 ng/dL, thyroid peroxidase &amp;lt;4, and thyroglobulin AB 19 IU/ml. Hoffmann’s Panel was confirmatory for this diagnosis reporting elevated levels of CPK 1509 U/L, LDH serum of 383 U/L and aldolase of 9.2 u/L. Ultrasound of the neck was non-contributory. Patient received antibiotic therapy for his skin infection and was also treated with IV Levothyroxine, which was prescribed orally upon discharge. Myopathy may be present in patients with longstanding untreated hypothyroidism in which pseudohypertrophy may occur, sometimes involving calf muscles (gastrocnemius). Hoffmann’s Syndrome is associated with changes in muscle fiber type from fast twitch type II to slow twitch type I and alteration of oxidative muscle enzyme activity with deceased calcium ATPase activity of fast twitch type II fibers leading to delayed relaxation. Biopsy of the affected muscle may reveal fiber necrosis, atrophy, hypertrophy with increased number of nuclei and increased connective tissue. Most cases of Hoffmann’s Syndrome improve after treatment with Levothyroxine, therefore, implementing this management on time can be curative. This case highlights the importance of considering this diagnosis when muscle dystrophies are studied and myopathic disorders are suspected, even in the absence of overt manifestations of hypothyroidism. Presentation: Thursday, June 15, 2023

Identification of mouse soleus muscle proteins altered in response to changes in gravity loading

Gravity-dependent physical processes strongly affect the ability of elderly people to maintain musculoskeletal health by reducing muscle atrophy and increasing bone mineral density, thereby increasing quality of life. A need therefore exists to identify molecules in the musculoskeletal system that are responsive to gravitational loading and to establish an objective indicator for the maintenance of healthy musculoskeletal systems. Here, we performed an integrated assessment of the results of soleus muscle proteomic analyses in three model mouse experiments under different gravity environments (hypergravity, hindlimb unloading, and spaceflight). Myl6b, Gpd1, Fbp2, Pvalb, and Actn3 were shown to be gravity-responsive muscle proteins, and alterations in the levels of these proteins indicated changes in muscle fiber type to slow-twitch type due to gravity loading. In addition, immunoblotting and enzyme-linked immunosorbent assays revealed that Pvalb levels in the sera of hindlimb-unloaded mice and osteoporosis patients were higher than in control subjects, suggesting that Pvalb levels might be useful to objectively evaluate soleus muscle atrophy and bone loss.

Musculoskeletal defects associated with myosin heavy chain‐embryonic loss of function are mediated by the YAP signaling pathway

Mutations in MYH3, the gene encoding the developmental myosin heavy chain‐embryonic (MyHC‐embryonic) skeletal muscle‐specific contractile protein, cause several congenital contracture syndromes. Among these, recessive loss‐of‐function MYH3 mutations lead to spondylocarpotarsal synostosis (SCTS), characterized by vertebral fusions and scoliosis. We find that Myh3 germline knockout adult mice display SCTS phenotypes such as scoliosis and vertebral fusion, in addition to reduced body weight, muscle weight, myofiber size, and grip strength. Myh3 knockout mice also exhibit changes in muscle fiber type, altered satellite cell numbers and increased muscle fibrosis. A mass spectrometric analysis of embryonic skeletal muscle from Myh3 knockouts identified integrin signaling and cytoskeletal regulation as the most affected pathways. These pathways are closely connected to the mechanosensing Yes‐associated protein (YAP) transcriptional regulator, which we found to be significantly activated in the skeletal muscle of Myh3 knockout mice. To test whether increased YAP signaling might underlie the musculoskeletal defects in Myh3 knockout mice, we treated these mice with CA3, a small molecule inhibitor of YAP signaling. This led to increased muscle fiber size, rescue of most muscle fiber type alterations, normalization of the satellite cell marker Pax7 levels, increased grip strength, reduced fibrosis, and decline in scoliosis in Myh3 knockout mice. Thus, increased YAP activation underlies the musculoskeletal defects seen in Myh3 knockout mice, indicating its significance as a key pathway to target in SCTS and other MYH3‐related congenital syndromes.

VEGF alleviates lower limb ischemia in diabetic mice by altering muscle fiber types.

Lower limb ischemia caused by diabetic foot (DF) is one of the most serious complications of diabetes. The therapeutic role of VEGF in DF is well documented. However, the mechanism for action of VEGF is still not clear. The present study aimed to explore the effects of VEGF-mediated skeletal muscle fiber type switch in angiogenesis and the treatment of DF. C57BL/6 mice housed in cages equipped with a voluntary running wheel were used to access VEGF protein level and citrate synthase activity (by ELISA) as well as muscle fiber type changes (by immunofluorescence) in the gastrocnemius muscle. C57BL/6 mice were fed on a high-fat diet for 6 weeks and then injected with streptozocin to induce diabetic lower limb ischemia model. Control adenovirus (Ad-GFP) or Ad-VEGF-GFP were then injected into the left gastrocnemius of the ischemic diabetic mice. Blood flow perfusion was examined by laser Doppler imaging at 1, 3, 7 and 14 days after adenovirus transduction. On day 14, all mice were anesthetized and sacrificed. VEGF expression levels, citrate synthase activity and muscle fiber type changes in the gastrocnemius muscle were assayed by ELISA and immunofluorescence analysis of myosin heavy chain IIa (MHCIIa) expression, respectively. Transwell assays were performed to determine whether VEGF-treated C2C12 myotubes played a role on tubule formation and migration of HUVECs. It was found that VEGF levels and citrate synthase activity were upregulated after voluntary exercise, along with the increased frequency of oxidized muscle fibers. Notably, adenovirus-mediated VEGF overexpression in the muscle also increased the frequency of oxidized (MHCIIa-positive) muscle fibers, enhanced citrate synthase activity and ameliorated lower limb ischemia in diabetic mice. VEGF treatment enhanced the phosphorylation of PI3K, Akt and AMPK (assayed by western blotting), as well as glucose consumption and metabolism (assayed by western blotting and glucose uptake assay), in the C2C12 myotubes. Interestingly, VEGF-treated C2C12 myotubes promoted the migration and tubule formation of HUVEC cells. The present findings suggest that skeletal muscle fiber conversion might be a potential approach for VEGF-mediated angiogenesis and disease treatment, which may provide new options for the prevention and treatment of DF.

Muscle Fiber Type Changes in Lumbrical Muscles at Early Stages of Chronic Nerve Compression.

Chronic nerve compression (CNC) neuropathy is a common disease in the clinic and provokes paraesthesia, or numbness at early stage. The changes in muscle fiber composition and motor nerve terminal morphology in distal muscles were studied in this study. A well-established CNC model was used to assess the changes in the muscles. Behaviors were measured by von Frey filament test. The myosin heavy chain isoforms and neuromuscular junctions (NMJs) were stained by immunofluorescence to show the muscle fiber types composition and motor nerve terminals morphologic changes in the flexor digitorum longus (FDL) and lumbrical muscle. The fiber cross-sectional areas of different muscle fiber types were measured. The small-fiber degeneration of cutaneous nerve fibers was examined by detecting the protein gene product 9.5 (PGP9.5) with immunofluorescence. At 2nd month after compression, the proportion of type I and type II B fibers was markedly decreased, and that of type II A fibers was increased in the lumbrical muscle. There was no significant change in composition of muscle fiber types in FDL and NMJ morphology of FDL and lumbrical muscles. Intra-epidermal nerve fibre density (IENFD) declined at 2nd month after the compression. Our study reveals the morphological changes of the FDL and lumbrical muscle at an early stage of CNC. These findings may be helpful to understand muscle damage and pathophysiological development of the nerve compression, and provide new evidence for early treatment of CNC.

OR-048 Effects and Mechanism of myokines in exercise mediated improving obesity rats skeletal muscle remodeling

Objective Aims From the year 2000, many experimental research data have indicated that skeletal muscle could express, synthesis and secrete multiple cytokines and polypeptides. The cytokines and polypeptides, not only regulate skeletal muscle growth, metabolism and motor function by paracrine/autocrine pathway, but also regulate functions of peripheral tissue and organs by endocrine pathway. Further researches proposed muscle as a secretory organ played a key role in mediating the health-promoting effects of physical activity and proteins expressed and released by skeletal muscle have been termed as myokines. Disorders of skeletal muscle endocrine function have related to the occurrence and development of multiple metabolic diseases, and myokines participate in obesity skeletal muscle remodeling. This study aims to investigate the expression changes of myokines and its effects in exercise mediated improving skeletal muscle remodeling on obesity mice, and explore the underlying mechanism of its functions.&#x0D; Methods Methods Five-week-old male Sprague-Dawley(SD) rats were randomly divided into a control group of 8 and a high-fat diet（HFD）group of 16. The control group was given normal food，while the HFD group were provided with high-fat diet for eight weeks and further divided into a sedentary HFD group and a treadmill running HFD group，each of 8. The exercise mice underwent 60 min treadmill running at 26 m/min each day，5 days/week for 8 weeks. Biochemical analyses, immune-histochemical, ELISA, RT-PCR and Western Blot methods were used to investigate multiple myokines expression changes and its mechanism.&#x0D; Results Results 1) Exercise significantly upregulated the expression of IL-15 in soleus and gastrocnemius muscle of obesity rats, indicating IL-15 could inhibit skeletal muscle endoplasmic reticulum stress and improve insulin sensitivity. 2) Exercise significantly inhibited the expression of myostatin (MSTN) in gastrocnemius muscle and mediated the changes of muscle fiber types. 3) Exercise markedly promoted the expression of apelin/APJ and angiogenesis function in obesity skeletal muscle. 4) Exercise upregulated skeletal muscle vascular endothelial growth factor B receptor expression and improved skeletal muscle ectopic lipid accumulation.&#x0D; Conclusions Conclusion Exercise regulates skeletal muscle myokines expression and secretion and have the effects on skeletal muscle fiber type changes, myofiber capillary density, glucose and lipid metabolism, thus improves the skeletal muscle remodeling and maintain body homeostasis.

Phytol Promotes the Formation of Slow-Twitch Muscle Fibers through PGC-1α/miRNA but Not Mitochondria Oxidation.

Phytol is a side chain of chlorophyll belonging to the side-chain double terpenoid. When animals consume food rich in chlorophyll, phytol can be broken down to phytanic acid after digestion. It was reported that feeding animals with different varieties and levels of forage could significant improve pH and marbling score of steer and lamb carcasses, but the internal mechanism for this is still not reported. The marbling score and pH of muscle was mainly determined by skeletal muscle fiber type, which is due to expression of different myosin heavy-chain (MHC) isoforms. Here, we provide evidence that phytol can indeed affect the diversity of muscle fiber types both in vitro and in vivo and demonstrate that phytol can increase the expression of MHC I (p < 0.05), likely by upgrading the expression of PPARδ, PGC-1α, and related miRNAs. This fiber-type transformation process may not be caused by activated mitochondrial metabolism but by the structural changes in muscle fiber types.

Modulation of Muscle Fiber Compositions in Response to Hypoxia via Pyruvate Dehydrogenase Kinase-1.

Muscle fiber-type changes in hypoxic conditions in accordance with pyruvate dehydrogenase kinase (Pdk)-1 and hypoxia inducible factor (Hif)-1α were investigated in rats. Hif-1α and its down-stream molecule Pdk-1 are well known for readily response to hypoxia. We questioned their roles in relation to changes in myosin heavy chain (MyHC) composition in skeletal muscles. We hypothesize that the level of Pdk-1 with respect to the level of Hif-1α determines MyHC composition of the muscle in rats in hypoxia. Young male rats were housed in a chamber maintained at 11.5% (for sustained hypoxia) or fluctuating between 11.5 and 20.8% (for intermittent hypoxia or IH) oxygen levels. Then, muscle tissues from the geniohyoid (GH), soleus, and anterior tibialis (TA) were obtained at the end of hypoxic conditionings. After both hypoxic conditionings, protein levels of Pdk-1 and Hif-1 increased in GH muscles. GH muscles in acute sustained hypoxia favor an anaerobic glycolytic pathway, resulting in an increase in glycolytic MyHC IIb protein-rich fibers while maintain original fatigue-resistant MyHC IIa protein in the fibers; thus, the numbers of IIa- and IIb MyHC co-expressing fibers increased. Exogenous Pdk-1 over-expression using plasmid vectors elevated not only the glycolytic MyHC IIb, but also IIx as well as IIa expressions in C2C12 myotubes in ambient air significantly. The increase of dual expression of IIa- and IIb MyHC proteins in fibers harvested from the geniohyoid muscle has a potential to improve endurance as shown in our fatigability tests. By increasing the Pdk-1/Hif-1 ratio, a mixed-type muscle could alter endurance within the innate characteristics of the muscle toward more fatigue resistant. We conclude that an increased Pdk-1 level in skeletal muscle helps maintain MyHC compositions to be a fatigue resistant mixed-type muscle.

Changes in muscle fiber type and expression of mRNA of myosin heavy chain isoforms in porcine muscle during pre- and postnatal development.

The purpose of this study is to elucidate developmental changes in muscle fiber type in the pig during pre- and postnatal development. For this purpose, we performed a histochemical analysis for myosin adenosine triphosphatase activity to assess muscle fiber type and determined abundances of messenger RNA (mRNA) of myosin heavy chain (MHC) isoforms. Samples of Longissimus dorsi (LD) muscle were taken from fetuses on day 90 of the fetal stage. Further, samples of LD, Rhomboideus and Biceps femoris (B. femoris) muscles were taken from pigs when they were 1, 12, 26, 45 or 75 days old. Expression of MHC 2b mRNA in the LD and the B. femoris muscles rapidly and considerably increased from the late fetal stage to the early postnatal stage and this increase was associated with the development of type 2b fibers at least in the LD muscle. As shown by the rapid and considerable changes in expression of MHC 2b mRNA, it seems that a certain plasticity of muscle fiber type still remains in this developmental stage.

Muscular Adaptations and Novel Magnetic Resonance Characterizations of Spinal Cord Injury

The spinal cord is highly complex, consisting of a specialized neural network that comprised both neuronal and non-neuronal cells. Any kind of injury and/or insult to the spinal cord leads to a series of damaging events resulting in motor and/or sensory deficits below the level of injury. As a result, muscle paralysis (or paresis) leading to muscle atrophy or shrinking of the muscle along with changes in muscle fiber type, and contractile properties have been observed. Traditionally, histology had been used as a gold standard to characterize spinal cord injury (SCI)-induced adaptation in spinal cord and skeletal muscle. However, histology measurements is invasive and cannot be used for longitudinal analysis. Therefore, the use of conventional magnetic resonance imaging (MRI) is promoted to be used as an alternative non-invasive method, which allows the repeated measurements over time and secures the safety against radiation by using radiofrequency pulse. Currently, many of pathological changes and adaptations occurring after SCI can be measured by MRI methods, specifically 3-dimensional MRI with the advanced diffusion tensor imaging technique. Both techniques have shown to be sensitive in measuring morphological and structural changes in skeletal muscle and the spinal cord.

Can Proinflammatory Cytokine Gene Expression Explain Multifidus Muscle Fiber Changes After an Intervertebral Disc Lesion?

Longitudinal case-controlled animal study. To investigate the effect of an intervertebral disc (IVD) lesion on the proportion of slow, fast, and intermediate muscle fiber types in the multifidus muscle in sheep, and whether muscle fiber changes were paralleled by local gene expression of the proinflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin 1-β. Structure and behavior of the multifidus muscle change in acute and chronic back pain, but the mechanisms are surprisingly poorly understood and the link between structure and behavior is tenuous. Although changes in muscle fiber types have the potential to unify the observations, the effect of injury on muscle fiber distribution has not been adequately tested, and understanding of possible mechanisms is limited. The L1-L2, L3-L4, and L5-L6 IVDs of 11 castrated male sheep received anterolateral lesions. Six control sheep underwent no surgical procedures. Multifidus muscle tissue was harvested at L4 for muscle fiber analysis using immunohistochemistry and L2 for cytokine analysis with polymerase chain reaction for local gene expression of TNF-α and interleukin-1β. The proportion of slow muscle fibers in multifidus was significantly less in the lesioned animals both ipsilateral and contralateral to the IVD lesion. The greatest reduction in slow fibers was in the deep medial muscle region. A greater prevalence of intermediate fibers on the uninjured side implies a delayed fiber-type transformation on that side. TNF-α gene expression in multifidus was greater on both sides in the lesion animals than in the muscle of control animals. Interleukin-1β was increased only on the injured side. These data provide evidence of muscle fiber changes after induction of an IVD lesion and a parallel increase in TNF-α expression. Proinflammatory cytokine changes provide a novel mechanism to explain behavioral and structural changes in multifidus. N/A.

Proteomics analysis of triceps muscle after C_7 nerve root rhizotomy in rats

Objective To investigate the protein expression changes in rat triceps muscle after C7 nerve root rhizotomy by two way fluorescence differential gel electrophoresis (2D-DICE),and to study the molecular mechanism of the triceps muscle's functional compensation. Methods C7 nerve root rhizotomy model was created on one side of the SD rats.The contralateral side did not undergo surgery and served as control.Equal amounts of triceps muscle were harvested from both sides after 1,2,3,6 and 12 weeks.The total proteins from the C7 rhizotomy side and from the normal side were examined by the 2D-DIGE technique.One way ANOVA was used to screen protein spots that differentiated with statistical significance.These protein spots were then identified by MALDI-TOF-MS or MS/MS. Results At different time points following C7 rhizotomy there were 38 protein spots detected with significant change.Among them 27 proteins were identified by MALDI-TOF-MS or MS/MS.11proteins belonged to functional groups associated with the energy metabolism of muscles.One week after rat C7 nerve root rhizotomy,5 protein spots of anaerobic metabolism enzyme down-regulated,then displayed an upregulation in their relative expression levels from 3rd to 12th week postoperatively.However 6 protein spots of aerobic oxidation enzymes down-regulated together with mitochondrial elongation factor Tu from 6th to 12th week.Conclusion The functional compensation of triceps after rat C7 nerve root rhizotomy may be associated with the changes in muscle fiber types,and the regulation of such a change may occur in the level of protein translation ofenergy metabolism enzyme. Key words: Proteomics; Peripheral nerves; Triceps brachii; 2D-DIGE

Reduction of type IIb myosin and IIB fibers in tibialis anterior muscle of mini‐muscle mice from high‐activity lines

Selective breeding of laboratory house mice (Mus domesticus) for high voluntary wheel running has generated four replicate lines that show an almost threefold increase in daily wheel-running distances as compared with four nonselected control lines. An unusual hindlimb "mini-muscle" phenotype (small muscles, increased mitochondrial enzyme levels, disorganized fiber distribution) has increased in frequency in two of the four replicate selected lines. The gene of major effect that accounts for this phenotype is an autosomal recessive that has been mapped to a 2.6335 Mb interval on MMU11, but not yet identified. This study examined the tibialis anterior muscle to determine whether changes in muscle fiber types could explain such modifications in muscle size and properties. Although selected and control lines did not exhibit systematic differences in the fiber types present in the tibialis anterior muscle, as assessed by electrophoresis of myosin heavy chains (MHC) and by histochemistry, mini-muscle mice lacked type IIB fibers and the corresponding MHCs. Mini-muscle tibialis show increased activities of hexokinase and citrate synthase compared with the normally sized muscles, likely the result of the modified fiber types in the muscle. The mini-muscle phenotype is the major means through which selective breeding for high wheel running has modified the functional capacities of the hindlimb muscles, as normally sized tibialis anterior muscles from control and selected lines did not show general differences in their enzymatic capacities, MHC profiles or fiber type composition, with the exception of an elevated hexokinase activity and a reduced GPa activity in the selected lines.

Increased oxidative properties of gastrocnemius in rats fed on a high-protein diet

It has been reported that a high-protein diet containing 30% protein partially prevents skeletal muscle fiber type changes (slow to fast) under atrophic conditions. No studies have examined the detailed effects of dietary protein on the oxidative properties of skeletal muscles. We examined the effects of a high-protein diet on the oxidative properties of rat gastrocnemius. Nineteen male Wistar rats (5 weeks old) were divided into the following groups: (1) control diet [15% protein (15P); n=6], (2) 25% protein (25P; n=6) and (3) 35% protein (35P, n=7). After 4 weeks of feeding, succinate dehydrogenase (SDH) staining and myosin isoforms were analyzed, along with the expression of the major transcription-related molecules for fast-to-slow fiber transition, i.e., peroxisome proliferator-activated receptor gamma coactivator (PGC) 1α and nuclear factor of activated T cells (NFAT) c1. SDH staining showed that the relative oxidative fiber content of the 35P group was significantly higher than that of the 15P group. The slow myosin heavy chain content in the 35P group was also higher than that in the 15P group. Western blotting analysis showed that the gastrocnemius of the 35P group contained significantly higher amounts of PGC1α and NFATc1 than that of the 15P group. We conclude that a high-protein diet, i.e., 35% protein, induces oxidative properties in rat gastrocnemius.

Androgen dependent seasonal changes in muscle fiber type in the dewlap neuromuscular system of green anoles

Green anoles ( Anolis carolinensis) possess two sexually dimorphic neuromuscular systems involved in reproductive behaviors. One controls extension of a red throat fan (dewlap), which males employ during courtship, and the other controls intromission of copulatory organs (hemipenes). Although seasonal changes in circulating androgens mediate both courtship and copulatory behaviors, testosterone has differential effects on the underlying neuromuscular morphology. The present experiments were designed to test whether changes in muscle fiber type correspond to seasonal and androgenic regulation of reproductive behaviors in gonadally intact males (Experiment 1) or castrated males treated with either testosterone propionate or vehicle (Experiment 2). Gonadally intact males housed in breeding environmental conditions had a higher percentage of fast oxidative glycolytic fibers in the dewlap muscle than non-breeding males, but no effect of season on copulatory fibers was detected. Interestingly, testosterone treatment increased the percentage of fast oxidative glycolytic dewlap fibers independent of season, suggesting that routine changes in this hormone may mediate fiber type in gonadally intact males. In contrast, testosterone manipulation had little to no effect on copulatory muscle fiber type, demonstrating that a change in this feature is not the primary mediator for seasonal changes in male copulatory behaviors.

Age-associated Decrease of Type IIA/B Human Skeletal Muscle Fibers

Elderly individuals often fall because of poor muscle strength and reduced balancing ability related to muscle aging. However, it is unclear whether changes in muscle fiber types contribute to poor strength or imbalance. We studied age- associated changes in human skeletal muscle fibers using muscle biopsy specimens taken from 65 male and female Chinese patients aged 17-96 years. The muscle specimens were cryosectioned with alkaline triphosphatase staining at pH 4.4, followed by image analysis. We analyzed morphologic observations and performed quantitative analyses of the number, size, and area percentage of different types of skeletal muscle fibers and connective tissues. Types IIA and IIB muscle fibers decreased with age in the area percentage, fiber number percentage, and mean fiber area, whereas Type I fibers increased in area and number but not in size. Morphologically, Type II fibers appeared smaller and flatter. Our findings suggest deterioration in muscle quality and balancing coordination in elderly patients. We provide data to help determine treatments for reversing muscle fiber changes and reducing the number of falls and related fractures in patients.