Area Of Individual Muscle Fibers Research Articles

Hyperphosphatemia Contributes to Skeletal Muscle Atrophy in Chronic Kidney Disease

Alterations in mineral metabolism involve changes in serum levels of phosphate as well as its key regulators, fibroblast growth factor 23 (FGF23), vitamin D, parathyroid hormone and klotho. Chronic Kidney Disease (CKD) is a public health epidemic that affects an estimated 37 million Americans, and is associated with elevated serum levels of phosphate (hyperphosphatemia) and FGF23 as well as various pathologies, such vascular calcification, cardiac hypertrophy, and skeletal muscle atrophy. We previously found that elevated FGF23 directly targets cardiac myocytes and contributes to cardiac hypertrophy. Furthermore, elevated phosphate has direct effects on smooth muscle cells and induces vascular calcification. In order to determine whether FGF23 and/or phosphate might also contribute to CKD-associated skeletal muscle atrophy, we tested if skeletal muscle cells can respond to elevations of FGF23 or phosphate. Since the skeletal muscle phenotype in CKD is not well characterized, we analyzed skeletal muscle on a functional, histological, and molecular level in two established mouse models of CKD. We studied the C2C12 skeletal muscle cell line during the differentiation process from myoblasts to myotubes as well as fully differentiated myotubes. Myoblast differentiation was induced with horse serum; cells were co-treated with 25 ng/ml FGF23 or 5 mM phosphate for 5 days, followed by the expression analysis of differentiation markers by qPCR. In a separate study, differentiated C2C12 myotubes were treated with FGF23 or phosphate for 24 hours, followed by the qPCR analysis of atrophy genes (atrogenes), including MT1, Murf1, and Fbox32. Furthermore, we studied mice with global deletion of collagen 4a3 (Col4a3-/-) and wildtype littermates at 10 weeks of age, as well as C57Bl/6 mice receiving an adenine-rich (0.2%) diet or regular chow for 12 weeks. In all mice we studied grip strength, hindlimb circumference by MRI, cross-sectional area of individual muscle fibers that were immuno-labeled with anti-laminin by fluorescence microscopy, and expression levels of atrogenes by qPCR. Phosphate but not FGF23 affected the differentiation process of C2C12 myoblasts and differentiated C2C12 myotubes. Phosphate treatments significantly reduced expression levels of MyoD and MyoG and increased expression of MT1. In both mouse models, grip strength and areas of hindlimb and myotubes were significantly reduced, and expression levels of MT1 and Murf1 were significantly elevated when compared to their controls. FGF23 does not target skeletal muscle cells suggesting that it cannot directly contribute to skeletal muscle atrophy. Elevated phosphate impairs myoblast differentiation and induces atrophy in vitro. We have characterized two CKD mouse models with associated skeletal muscle atrophy, where we will now determine the contribution of hyperphosphatemia to the injury process. Pharmacological approaches targeting phosphate uptake or excretion, or phosphate's direct actions on tissues might alleviate various CKD-associated pathologies.

Quantitative Analysis of Muscle Histologic Method in Rodent Facial Nerve Injury

To describe denervation features of facial musculature following facial nerve injury in a rodent model. Six Wistar-Hannover rats underwent unilateral transection and immediate repair of the facial nerve. After 8 weeks, muscular bundles consisting of dilator naris and levator labii superioris from both sides were analyzed for mean muscle cell diameter and the percentage of muscle cell cross-sectional area using image processing software. The atrophic features of facial muscles were quantified and compared with the contralateral, healthy side of the face. Weekly postoperative whisking assessment demonstrated the anticipated course of recovery. We observed significant differences between the normal side and the manipulated side, respectively, in the percentage of muscle specimen cross-sectional area attributable to muscle cell profiles (57% vs 29%; P = .006) and total fiber counts (1346 vs 794; P = .02). The mean cross-sectional area of individual muscle fibers was higher on the normal side (1129 vs 928 μm2; P = .39); however, this difference was statistically nonsignificant. The objective, quantitative measures of muscle microstructure used in this report provide a valuable point of comparison for whisking function and electrophysiologic measures and can be used in future studies to assess muscle atrophic features associated with facial nerve injury and repair techniques.

Resistance exercise increases leg muscle protein synthesis and mTOR signalling independent of sex

Sex differences are evident in human skeletal muscle as the cross-sectional area of individual muscle fibres is greater in men than in women. We have recently shown that resistance exercise stimulates mammalian target of rapamycin (mTOR) signalling and muscle protein synthesis in humans during early post-exercise recovery. Therefore, the aim of this study was to determine if sex influences the muscle protein synthesis response during recovery from resistance exercise. Seventeen subjects, nine male and eight female, were studied in the fasted state before, during and for 2 h following a bout of high-intensity leg resistance exercise. Mixed muscle protein fractional synthetic rate was measured using stable isotope techniques and mTOR signalling was assessed by immunoblotting from repeated vastus lateralis muscle biopsy samples. Post-exercise muscle protein synthesis increased by 52% in the men and by 47% in the women (P < 0.05) and was not different between groups (P > 0.05). Akt phosphorylation increased in both groups at 1 h post-exercise (P < 0.05) and returned to baseline during 2 h post-exercise with no differences between groups (P > 0.05). Phosphorylation of mTOR and its downstream effector S6K1 increased significantly and similarly between groups during post-exercise recovery (P < 0.05). eEF2 phosphorylation decreased at 1- and 2 h post-exercise (P < 0.05) to a similar extent in both groups. The contraction-induced increase in early post-exercise mTOR signalling and muscle protein synthesis is independent of sex and appears to not play a role in the sexual dimorphism of leg skeletal muscle in young men and women.

Myoglobin concentration in skeletal muscle fibers of chronic heart failure patients

The purpose of this study was to determine the myoglobin concentration in skeletal muscle fibers of chronic heart failure (CHF) patients and to calculate the effect of myoglobin on oxygen buffering and facilitated diffusion. Myoglobin concentration, succinate dehydrogenase (SDH) activity, and cross-sectional area of individual muscle fibers from the vastus lateralis of five control and nine CHF patients were determined using calibrated histochemistry. CHF patients compared with control subjects were similar with respect to myoglobin concentration: type I fibers 0.69 +/- 0.11 mM (mean +/- SD), type II fibers 0.52 +/- 0.07 mM in CHF vs. type I fibers 0.70 +/- 0.09 mM, type II fibers 0.49 +/- 0.07 mM in control, whereas SDH activity was significantly lower in CHF in both fiber types (P < 0.01). The myoglobin concentration in type I fibers was higher than in type II fibers (P < 0.01). Consequently, the oxygen buffering capacity, calculated from myoglobin concentration/SDH activity was increased in CHF: type I fibers 11.4 +/- 2.1 s, type II fibers 13.6 +/- 3.9 s in CHF vs. type I fibers 7.8 +/- 0.9 s, type II fibers 7.5 +/- 1.0 s in control, all P < 0.01). The calculated extracellular oxygen tension required to prevent core anoxia (Po2(crit)) in muscle fibers was similar when controls were compared with patients in type I fibers 10.3 +/- 0.9 Torr in CHF and 11.5 +/- 3.3 Torr in control, but was lower in type II fibers of patients 6.1 +/- 2.8 Torr in CHF and 14.7 +/- 6.2 Torr in control, P < 0.01. The lower Po2(crit) of type II fibers may facilitate oxygen extraction from capillaries. Reduced exercise tolerance in CHF is not due to myoglobin deficiency.

Effects of age and breed of beef bulls on growth parameters, carcass and muscle characteristics

AbstractThe effects of age and breed on growth parameters, carcass and muscle characteristics of bulls, slaughtered at 15, 19 and 24 months of age, were analysed in four French breeds: Aubrac (AU), Charolais (CH), Limousin (LI), and Salers (SA). Muscle characteristics were determined in three muscles: longissimus thoracis (LT), semitendinosus (ST) and triceps brachii (TB). They included: (1) the % frequency, cross-sectional area and % area of fibre types, which were classified according to the contractile nature of the fibres and their metabolic properties (SO slow oxidative, FOG fast oxidative glycolytic and FG fast glycolytic); (2) the isocitrate dehydrogenase (ICDH) and lactate dehydrogenase (LDH) activities, representative of oxidative and glycolytic metabolism respectively; and (3) the total and insoluble collagen contents.In the four breeds, the average daily gain and the food efficiency decreased with age (P &lt; 0·001). The carcass characteristics (muscle, fat and bone weights) increased with age (P &lt; 0·001). The increase of muscle carcass weight with slaughter age was in parallel with the increase in cross-sectional area of individual muscle fibres. Oxidative fibre (SO and FOG) areas increased more between 15 and 24 months than glycolytic fibre (FG) area. Differences between muscles in increases in areas of muscle fibres were consistent: the increase was greater for TB than ST and LT. The muscles studied became more slow and more oxidative above 19 months of age, as evidenced by the fact that the SO % frequency (P &lt; 0·001) and % area (P &lt; 0·001) and ICDH activity (P &lt; 0·05) increased, and LDH activity decreased (P &lt; 0·01). Insoluble collagen content decreased between 15 and 19 months (P &lt; 0·001), and both total (P &lt; 0·01) and insoluble (P &lt; 0·001) collagen contents increased from 19 months.So carcass characteristics were modified between 15 and 24 months, and muscle characteristics were especially modified from 19 months of age. In addition, differences in slaughter data between breeds were clear and consistent, whereas those of muscle characteristics were few and not consistent.

Morphological characteristics of skeletal muscles in relation to gender.

The aim of this study was to ascertain whether there are gender-related differences in the morphological characteristics of the soleus and tibialis anterior muscles in young adult and old Fischer 344/Brown Norway F1 rats. We tested 1) whether there was a gender-related difference between the fiber type composition of these muscles, and 2) whether the cross-sectional area of individual muscle fibers demonstrated gender-associated differences, fibers from males being larger than fibers from females. Gender differences were not found in the fiber type composition of the soleus and tibialis anterior muscles, but were present in the single skeletal fiber cross-sectional area of the tibialis anterior muscle. The cross-sectional area of type I fibers in females was greater than that in males at both 12 (16%) and 30 (5%) months of age. In contrast, the cross-sectional area of type Ila fibers of 12-month-old males was larger than that of 12-month-old females. No significant differences between genders were found for the cross-sectional area of type Ilb fibers in either age group. In the soleus muscle, 30-month-old males had larger single fiber cross-sectional areas of both fiber types I and lIa. At 12 months of age, type I fibers from females were larger than those from males. Our findings indicate that gender-related differences exist in the size of individual skeletal fibers from the soleus and tibialis anterior muscles and that they may influence metabolism and the adaptive response to rehabilitation programs.

Comparative analysis of fiber‐type composition in the iliofibularis muscle of phrynosomatid lizards (Squamata)

The lizard family Phrynosomatidae comprises three subclades: the closely related sand and horned lizards, and their relatives the Sceloporus group. This family exhibits great variation in ecology, behavior, and general body plan. Previous studies also show that this family exhibits great diversity in locomotor performance abilities; as measured on a high-speed treadmill, sand lizards are exceptionally fast sprinters, members of the Sceloporus group are intermediate, and horned lizards are slowest. These differences are paralleled by differences in relative hindlimb span. To determine if muscle fiber-type composition also varies among the three subclades, we examined the iliofibularis (IF), a hindlimb muscle used in lizard locomotion, in 11 species of phrynosomatid lizards. Using histochemical assays for myosin ATPase, an indicator of fast-twitch capacity, and succinic dehydrogenase, denoting oxidative capacity, we classified fiber types into three categories based on existing nomenclature: fast-twitch glycolytic (FG), fast-twitch oxidative-glycolytic (FOG), and slow-twitch oxidative (SO). Sand lizards have a high proportion of FG fibers (64–70%) and a low proportion of FOG fibers (25–33%), horned lizards are the converse (FG fibers 25–31%, FOG fibers 56–66%), and members of the Sceloporus group are intermediate for both FG (41–48%) and FOG (42–45%) content. Hence, across all 11 species %FOG and %FG are strongly negatively correlated. Analysis with phylogenetically independent contrasts indicate that this negative relationship is entirely attributable to the divergence between sand and horned lizards. The %SO also varies among the three subclades. Results from conventional nested ANCOVA (with log body mass as a covariate) indicate that the log mean cross-sectional area of individual muscle fibers differs among species and is positively correlated with body mass across species, but does not differ significantly among subclades. The log cross-sectional area of the IF varies among species, but does not vary among subclades. Conversely, the total thigh muscle cross-sectional area does not vary among species, but does vary among subclades; horned lizards have slimmer thighs. Muscle fiber-type composition appears to form part of a coadapted suite of traits, along with relative limb and muscle sizes, that affect the locomotor abilities of phrynosomatid lizards. J. Morphol. 250:265–280, 2001. © 2001 Wiley-Liss, Inc.

The effects of age at slaughter, genotype and finishing system on the biochemical properties, muscle fibre type characteristics and eating quality of bull beef from suckled calves

AbstractMuscle fibre characteristics and biochemical properties of muscle recovered from young bulls of two genotypes (Aberdeen Angus × and Charolais ×), reared on two different diets (silage-based and barley-based) and slaughtered at varying ages between 10 and 19 months of age were established. These analyses were restricted to samples ofm. longissimus lumborum (LI) recovered at 48 h post mortem, vacuum packed and stored at 2°C for 14 days. Biochemical measurements included intramuscular fat content, intramuscular collagen content and its solubility, haem pigment concentration, sarcomere length and myofibril fragmentation. Muscle fibre type was classified according to the contractile nature of thefibres and their metabolic properties.Intramuscular fat content increased (P &lt; 0·01) with age at slaughter and at a fixed age was greater for Angus × than Charolais × bulls (211·5 v. 295·8 mg/g dry matter, P &lt; 0·01). Total intramuscular collagen and its solubility tended to decrease with age (P &lt; 0·01). Differences in haem pigment concentration in samples of LI were detected between genotype (3·99 v. 3·59 mg/g for Angus × and Charolais × bulls; P × 0·01) and diet (3·97 v. 3·62 mg/g for bulls given barley and silage; P &lt; 0·01), and increased with age at slaughter (P &lt; 0·01). There was a significant increase in eye muscle cross-sectional area with increasing slaughter date (P &lt; 0·01) and this was paralleled by an increase in cross-sectional area of individual muscle fibres (P &lt; 0·001). Differences in cross-sectional area of individual muscle fibres between genotype and diet were small and inconsistent. Charolais × bulls had a greater percentage area of fast twitch glycolytic fibres than Angus × bulls (54·3 v. 49·3%; P &lt; 0·01) and a smaller percentage area of slow twitch oxidative fibres (15·8 v. 18·9%; P &lt; 0·05). Beef tenderness was positively correlated (r = 0·48; P &lt; 0·01) with the frequency of slow twitch oxidative fibres and negatively correlated (r = -0·38; P &lt; 0·05) with the frequency offast twitch glycolytic fibres.

Capillarity and diffusion distances in skeletal muscles in birds

Tissue capillarity and diffusion distances were determined for red and white skeletal muscles of adult birds ranging in mass from 10.8 to 6200 g. In addition, literature values for capillarity and diffusion distances in skeletal muscles of mammals were incorporated into the data set. Muscle mass was closely coupled to body mass. However, no significant allometric relations were found for any of the other variables measured. Number of capillaries per fiber was not correlated with cross sectional area of individual muscle fibers. Thus, capillary density decreased in a hyperbolic manner against fiber area and diffusion distance decreased in a hyperbolic manner against the number of capillaries per muscle fiber. Red muscles had significantly higher numbers of capillaries per fiber and significantly shorter diffusion distances than did white muscles. The patterns for tissue capillarity and diffusion distances in avian muscle reported here are similar to values reported previously for mammalian muscles. In both taxonomic groups capillarity and diffusion distances are independent of body mass. In addition, diffusion distances are characteristic of capillaries distributed in random arrays through the muscle cross section.

SDH activity and cross-sectional area of muscle fibers in cat diaphragm.

The succinate dehydrogenase (SDH) activity and cross-sectional area of individual muscle fibers in the cat diaphragm were quantified using a computerized image-processing system. The population distributions of fiber-SDH activities and cross-sectional areas showed considerable range within each diaphragm. Despite an overlap in the distribution of SDH activities between fast-twitch (i.e., those staining darkly for myosin ATPase) and slow-twitch (i.e., those staining lightly for myosin ATPase) fibers, differences between the two populations of fibers were observed. Fast-twitch fibers generally had lower SDH activities and greater cross-sectional areas than slow-twitch fibers. However, the range of SDH activities and cross-sectional areas of fast-twitch fibers was much greater than in slow-twitch fibers. The population distributions of SDH activities and cross-sectional areas of both fast- and slow-twitch fibers were unimodal. The unimodal distribution of SDH activities in fast-twitch fibers suggested that these fibers could not be clearly subdivided into two groups based on differences in their oxidative capacity (i.e., high- and low-oxidative fibers). These results were discussed in the context of the classification of fast-twitch glycolytic and fast-twitch oxidative-glycolytic fibers using qualitative histochemistry. In addition, the functional significance of the unimodal distribution of oxidative capacities in fast-twitch fibers was discussed in relation to the distribution of fatigue properties in fast-twitch motor units. A significant negative correlation between cross-sectional area and SDH activity in both fast- and slow-twitch fibers was also observed. The importance of this negative correlation in relationship to the diffusion of energy substrates for oxidative metabolism was discussed.

Skeletal muscle fiber splitting induced by weight-lifting exercise in cats.

Adult skeletal muscle hypertrophy induced by exercise has been thought to be exclusively related to an increase in cross-section area of individual muscle fibers and not to an increase in the number of muscle fibers. Recent experiments using surgical intervention to cause muscle overload have induced an increase in fiber numbers; however, the muscle also exhibited pathological alterations. The purpose of this study was to determine if an exercise regimen also induced hyperplasia. Cats were trained to lift weights with their right forelimb to receive a food reward. After 19-46 weeks of training, the flexor carpi radialis muscle (FCR) was removed and prepared for histochemical examination. The total number of muscle fibers of the right exercised FCR increased significantly (19.3%) when compared with that of the unexercised left FCR (p less than 0.05). This increase was found to be due to muscle fiber splitting.

The Electrical and Mechanical Responses of the Prothoracic Flexor Tibialis Muscle of the Stick Insect, Carausius Morosus Br

ABSTRACT The prothoracic flexor tibialis muscle of Carausius morosus consists of two lateral rows of pinnately arranged muscle units. Motor nerve endings of the ‘Doyére-cone’ type are distributed at intervals of approximately 60 p along each fibre. Each motor ending is probably innervated by two axons. Two types of responses have been found in the muscle fibres: (i) ‘fast’ electrical responses resembling the action potential of vertebrate muscles, associated with twitch-type contractions of the fibres; (ii) ‘slow’ readily facilitating responses resembling end-plate potentials, associated with slow, smooth contractions of the muscle, and with the maintenance of tonus. There is no evidence of peripheral motor inhibition. The muscle bathed in haemolymph is capable of developing a tetanus tension of 800 g./cm.2 cross-sectional area of individual muscle fibres. The tetanus : twitch ratio is over 25:1. Pharmacological substances which affect excitable tissues of other animals have no effect on the fast response. Progressively lowered temperatures lengthen the time course and reduce the amplitude of the fast response, but an active membrane response remains at 50 C. Refractoriness is evident near the peak of the fast response. The junctional potentials will summate if sufficiently close in time. It is suggested that the process underlying the fast response in Carausius is similar to that in the locust and in vertebrates ; but neuromuscular transmission does not appear to be cholinergic.