Influence of Physical Exercise on the Rehabilitation of Volumetric Muscle Loss Injury Reconstructed with Autologous Adipose Tissue.

BACKGROUND: In volumetric muscle loss (VML) injuries, spontaneous muscle regeneration capacity is limited. The implantation of autologous adipose tissue in the affected area is an option to treat these lesions; however, the effectiveness of this therapy alone is insufficient for a complete recovery of the damaged muscle. This study examined the influence of treadmill exercise on the rehabilitation of VML injuries reconstructed with autologous adipose tissue, as a strategy to counteract the limitations of spontaneous regeneration observed in these injuries. METHODS: Forty adult male Wistar rats were divided into eight groups of five individuals each: normal control (NC), regenerative control (RC), VML control (VML), VML injury reconstructed with fresh autologous adipose tissue (FAT), exercise-rehabilitated control (RNC), exercise-rehabilitated regenerative control (RRC), exercise-rehabilitated VML injury (RVML), and exercise-rehabilitated VML injury reconstructed with fresh autologous adipose tissue (RFAT). Histological and histochemical staining techniques were used for the analysis of structural features and histomorphometric parameters of the tibialis anterior muscle. Grip strength tests were conducted to assess muscle force. RESULTS: Exercise rehabilitation decreased the proportion of disoriented fibers in RFAT vs. FAT group. The percentage of fibrosis was significantly higher in FAT and RFAT groups versus NC and RNC groups but did not vary significantly between FAT and RFAT groups. Overall, muscle grip strength and fiber size increased significantly in the exercise-rehabilitated groups compared to control groups. CONCLUSIONS: To conclude, rehabilitation with physical exercise tended to normalize the process of muscle repair in a model of VML injury reconstructed with fresh autologous adipose tissue, but it did not reduce the intense fibrosis associated with these injuries.

Muscle plasticity is influenced by renal function and caloric intake through the FGF23-vitamin D axis.

Skeletal muscle, the main metabolic engine in the body of vertebrates, is endowed with great plasticity. The association between skeletal muscle plasticity and two highly prevalent health problems: renal dysfunction and obesity, which share etiologic links as well as many comorbidities, is a subject of great relevance. It is important to know how these alterations impact on the structure and function of skeletal muscle because the changes in muscle phenotype have a major influence on the quality of life of the patients. This literature review aims to discuss the influence of a nontraditional axis involving kidney, bone, and muscle on skeletal muscle plasticity. In this axis, the kidneys play a role as the main site for vitamin D activation. Renal disease leads to a direct decrease in 1,25(OH)2-vitamin D, secondary to reduction in renal functional mass, and has an indirect effect, through phosphate retention, that contributes to stimulate fibroblast growth factor 23 (FGF23) secretion by bone cells. FGF23 downregulates the renal synthesis of 1,25(OH)2-vitamin D and upregulates its metabolism. Skeletal production of FGF23 is also regulated by caloric intake: it is increased in obesity and decreased by caloric restriction, and these changes impact on 1,25(OH)2-vitamin D concentrations, which are decreased in obesity and increased after caloric restriction. Thus, both phosphate retention, that develops secondary to renal failure, and caloric intake influence 1,25(OH)2-vitamin D that in turn plays a key role in muscle anabolism.

Increased 1,25(OH)2-Vitamin D Concentrations after Energy Restriction Are Associated with Changes in Skeletal Muscle Phenotype.

The influence of energy restriction (ER) on muscle is controversial, and the mechanisms are not well understood. To study the effect of ER on skeletal muscle phenotype and the influence of vitamin D, rats (n = 34) were fed a control diet or an ER diet. Muscle mass, muscle somatic index (MSI), fiber-type composition, fiber size, and metabolic activity were studied in tibialis cranialis (TC) and soleus (SOL) muscles. Plasma vitamin D metabolites and renal expression of enzymes involved in vitamin D metabolism were measured. In the ER group, muscle weight was unchanged in TC and decreased by 12% in SOL, but MSI increased in both muscles (p < 0.0001) by 55% and 36%, respectively. Histomorphometric studies showed 14% increase in the percentage of type IIA fibers and 13% reduction in type IIX fibers in TC of ER rats. Decreased size of type I fibers and reduced oxidative activity was identified in SOL of ER rats. An increase in plasma 1,25(OH)2-vitamin D (169.7 ± 6.8 vs. 85.4 ± 11.5 pg/mL, p < 0.0001) with kidney up-regulation of CYP27b1 and down-regulation of CYP24a1 was observed in ER rats. Plasma vitamin D correlated with MSI in both muscles (p < 0.001), with the percentages of type IIA and type IIX fibers in TC and with the oxidative profile in SOL. In conclusion, ER preserves skeletal muscle mass, improves contractile phenotype in phasic muscles (TC), and reduces energy expenditure in antigravity muscles (SOL). These beneficial effects are closely related to the increases in vitamin D secondary to ER.

Locomotor muscle fibre heterogeneity and metabolism in the fastest large-bodied rorqual: the fin whale (Balaenoptera physalus).

From a terrestrial ancestry, the fin whale (Balaenoptera physalus) is one of the largest animals on Earth with a sprinter anti-predator strategy, and a characteristic feeding mode, lunge feeding, which involves bouts of high-intensity muscle activity demanding high metabolic output. We investigated the locomotor muscle morphology and metabolism of this cetacean to determine whether its muscle profile (1) explains this unique swimming performance and feeding behaviour, (2) is or is not homogeneous within the muscle, and (3) predicts allometric variations inherent to an extreme body size. A predominantly fast-glycolytic phenotype characterized the fin whale locomotor muscle, composed of abundant fast-twitch (type IIA) fibres with high glycolytic potential, low oxidative capacity, relatively small size, and reduced number of capillaries. Compared with superficial areas, deep regions of this muscle exhibited a slower and more oxidative profile, suggesting a division of labour between muscle strata. As expected, the fin whale locomotor muscle only expressed the two slowest myosin heavy chain isoforms (I and IIA). However, it displayed anaerobic (glycolytic) and aerobic (lipid-based metabolism) capabilities higher than would be predicted from the allometric perspective of its extreme body size. Relationships between muscle metabolism and body mass were fibre-type specific. The 'sprinter' profile of the fin whale swimming muscle, particularly of its superficial compartment, supports physiological demands during both high-speed swimming and the lunge, when energy expenditure reaches maximal or supramaximal levels. Comparatively, the slower and more oxidative profile of the deep compartment of this muscle seems to be well designed for sustained, low-intensity muscle activity during routine swimming.

Obesity-induced discrepancy between contractile and metabolic phenotypes in slow- and fast-twitch skeletal muscles of female obese Zucker rats.

A clear picture of skeletal muscle adaptations to obesity and related comorbidities remains elusive. This study describes fiber-type characteristics (size, proportions, and oxidative enzyme activity) in two typical hindlimb muscles with opposite structure and function in an animal model of genetic obesity. Lesser fiber diameter, fiber-type composition, and histochemical succinic dehydrogenase activity (an oxidative marker) of muscle fiber types were assessed in slow (soleus)- and fast (tibialis cranialis)-twitch muscles of obese Zucker rats and compared with age (16 wk)- and sex (females)-matched lean Zucker rats (n = 16/group). Muscle mass and lesser fiber diameter were lower in both muscle types of obese compared with lean animals even though body weights were increased in the obese cohort. A faster fiber-type phenotype also occurred in slow- and fast-twitch muscles of obese rats compared with lean rats. These adaptations were accompanied by a significant increment in histochemical succinic dehydrogenase activity of slow-twitch fibers in the soleus muscle and fast-twitch fiber types in the tibialis cranialis muscle. Obesity significantly increased plasma levels of proinflammatory cytokines but did not significantly affect protein levels of peroxisome proliferator-activated receptors PPARγ or PGC1α in either muscle. These data demonstrate that, in female Zucker rats, obesity induces a reduction of muscle mass in which skeletal muscles show a diminished fiber size and a faster and more oxidative phenotype. It was noteworthy that this discrepancy in muscle's contractile and metabolic features was of comparable nature and extent in muscles with different fiber-type composition and antagonist functions.NEW & NOTEWORTHY This study demonstrates a discrepancy between morphological (reduced muscle mass), contractile (shift toward a faster phenotype), and metabolic (increased mitochondrial oxidative enzyme activity) characteristics in skeletal muscles of female Zucker fatty rats. It is noteworthy that this inconsistency was comparable (in nature and extent) in muscles with different structure and function.

Mangiferin protects against adverse skeletal muscle changes and enhances muscle oxidative capacity in obese rats.

Obesity-related skeletal muscle changes include muscle atrophy, slow-to-fast fiber-type transformation, and impaired mitochondrial oxidative capacity. These changes relate with increased risk of insulin resistance. Mangiferin, the major component of the plant Mangifera indica, is a well-known anti-inflammatory, anti-diabetic, and antihyperlipidemic agent. This study tested the hypothesis that mangiferin treatment counteracts obesity-induced fiber atrophy and slow-to-fast fiber transition, and favors an oxidative phenotype in skeletal muscle of obese rats. Obese Zucker rats were fed gelatin pellets with (15 mg/kg BW/day) or without (placebo group) mangiferin for 8 weeks. Lean Zucker rats received the same gelatin pellets without mangiferin and served as non-obese and non-diabetic controls. Lesser diameter, fiber composition, and histochemical succinic dehydrogenase activity (an oxidative marker) of myosin-based fiber-types were assessed in soleus and tibialis cranialis muscles. A multivariate discriminant analysis encompassing all fiber-type features indicated that obese rats treated with mangiferin displayed skeletal muscle phenotypes significantly different compared with both lean and obese control rats. Mangiferin significantly decreased inflammatory cytokines, preserved skeletal muscle mass, fiber cross-sectional size, and fiber-type composition, and enhanced muscle fiber oxidative capacity. These data demonstrate that mangiferin attenuated adverse skeletal muscle changes in obese rats.

Skeletal muscle adaptations and muscle genomics of performance horses.

Skeletal muscles in horses are characterised by specific adaptations, which are the result of the natural evolution of the horse as a grazing animal, centuries of selective breeding and the adaptability of this tissue in response to training. These adaptations include an increased muscle mass relative to body weight, a great locomotor efficiency based upon an admirable muscle-tendon architectural design and an adaptable fibre-type composition with intrinsic shortening velocities greater than would be predicted from an animal of comparable body size. Furthermore, equine skeletal muscles have a high mitochondrial volume that permits a higher whole animal aerobic capacity, as well as large intramuscular stores of energy substrates (glycogen in particular). Finally, high buffer and lactate transport capacities preserve muscles against fatigue during anaerobic exercise. Many of these adaptations can improve with training. The publication of the equine genome sequence in 2009 has provided a major advance towards an improved understanding of equine muscle physiology. Equine muscle genomics studies have revealed a number of genes associated with elite physical performance and have also identified changes in structural and metabolic genes following exercise and training. Genes involved in muscle growth, muscle contraction and specific metabolic pathways have been found to be functionally relevant for the early performance evaluation of elite athletic horses. The candidate genes discussed in this review are important for a healthy individual to improve performance. However, muscle performance limiting conditions are widespread in horses and many of these conditions are also genetically influenced.

High-phosphorus diet maximizes and low-dose calcitriol attenuates skeletal muscle changes in long-term uremic rats.

Although disorders of mineral metabolism and skeletal muscle are common in chronic kidney disease (CKD), their potential relationship remains unexplored. Elevations in plasma phosphate, parathyroid hormone, and fibroblastic growth factor 23 together with decreased calcitriol levels are common features of CKD. High-phosphate intake is a major contributor to progression of CKD. This study was primarily aimed to determine the influence of high-phosphate intake on muscle and to investigate whether calcitriol supplementation counteracts negative skeletal muscle changes associated with long-term uremia. Proportions and metabolic and morphological features of myosin-based muscle fiber types were assessed in the slow-twitch soleus and the fast-twitch tibialis cranialis muscles of uremic rats (5/6 nephrectomy, Nx) and compared with sham-operated (So) controls. Three groups of Nx rats received either a standard diet (0.6% phosphorus, Nx-Sd), or a high-phosphorus diet (0.9% phosphorus, Nx-Pho), or a high-phosphorus diet plus calcitriol (10 ng/kg 3 day/wk ip, Nx-Pho + Cal) for 12 wk. Two groups of So rats received either a standard diet or a high-phosphorus diet (So-Pho) over the same period. A multivariate analysis encompassing all fiber-type characteristics indicated that Nx-Pho + Cal rats displayed skeletal muscle phenotypes intermediate between Nx-Pho and So-Pho rats and that uremia-induced skeletal muscle changes were of greater magnitude in Nx-Pho than in Nx-Sd rats. In uremic rats, treatment with calcitriol preserved fiber-type composition, cross-sectional size, myonuclear domain size, oxidative capacity, and capillarity of muscle fibers. These data demonstrate that a high-phosphorus diet potentiates and low-dose calcitriol attenuates adverse skeletal muscle changes in long-term uremic rats.

Slow- and fast-twitch hindlimb skeletal muscle phenotypes 12 wk after ⠚ nephrectomy in Wistar rats of both sexes.

This study describes fiber-type adaptations in hindlimb muscles, the interaction of sex, and the role of hypoxia on this response in 12-wk ⅚ nephrectomized rats (Nx). Contractile, metabolic, and morphological features of muscle fiber types were assessed in the slow-twitch soleus and the fast-twitch tibialis cranialis muscles of Nx rats, and compared with sham-operated controls. Rats of both sexes were considered in both groups. A slow-to-fast fiber-type transformation occurred in the tibialis cranialis of Nx rats, particularly in males. This adaptation was accomplished by impaired oxidative capacity and capillarity, increased glycolytic capacity, and no changes in size and nuclear density of muscle fiber types. An oxidative-to-glycolytic metabolic transformation was also found in the soleus muscle of Nx rats. However, a modest fast-to-slow fiber-type transformation, fiber hypertrophy, and nuclear proliferation were observed in soleus muscle fibers of male, but not of female, Nx rats. Serum testosterone levels decreased by 50% in male but not in female Nx rats. Hypoxia-inducible factor-1α protein level decreased by 42% in the tibialis cranialis muscle of male Nx rats. These data demonstrate that 12 wk of Nx induces a muscle-specific adaptive response in which myofibers do not change (or enlarge minimally) in size and nuclear density, but acquire markedly different contractile and metabolic characteristics, which are accompanied by capillary rarefaction. Muscle function and sex play relevant roles in these adaptations.

Adaptive functional specialisation of architectural design and fibre type characteristics in agonist shoulder flexor muscles of the llama, Lama glama.

Like other camelids, llamas (Lama glama) have the natural ability to pace (moving ipsilateral limbs in near synchronicity). But unlike the Old World camelids (bactrian and dromedary camels), they are well adapted for pacing at slower or moderate speeds in high-altitude habitats, having been described as good climbers and used as pack animals for centuries. In order to gain insight into skeletal muscle design and to ascertain its relationship with the llama's characteristic locomotor behaviour, this study examined the correspondence between architecture and fibre types in two agonist muscles involved in shoulder flexion (M. teres major - TM and M. deltoideus, pars scapularis - DS and pars acromialis - DA). Architectural properties were found to be correlated with fibre-type characteristics both in DS (long fibres, low pinnation angle, fast-glycolytic fibre phenotype with abundant IIB fibres, small fibre size, reduced number of capillaries per fibre and low oxidative capacity) and in DA (short fibres, high pinnation angle, slow-oxidative fibre phenotype with numerous type I fibres, very sparse IIB fibres, and larger fibre size, abundant capillaries and high oxidative capacity). This correlation suggests a clear division of labour within the M. deltoideus of the llama, DS being involved in rapid flexion of the shoulder joint during the swing phase of the gait, and DA in joint stabilisation during the stance phase. However, the architectural design of the TM muscle (longer fibres and lower fibre pinnation angle) was not strictly matched with its fibre-type characteristics (very similar to those of the postural DA muscle). This unusual design suggests a dual function of the TM muscle both in active flexion of the shoulder and in passive support of the limb during the stance phase, pulling the forelimb to the trunk. This functional specialisation seems to be well suited to a quadruped species that needs to increase ipsilateral stability of the limb during the support phase of the pacing gait. Compared with other species, llama skeletal muscles are well suited for greater force generation combined with higher fatigue resistance during exercise. These characteristics are interpreted as being of high adaptive value, given the llama's habitat and its use as a pack animal.

Alterations in oxidative gene expression in equine skeletal muscle following exercise and training.

Intense selection for elite racing performance in the Thoroughbred horse (Equus caballus) has resulted in a number of adaptive physiological phenotypes relevant to exercise; however, the underlying molecular mechanisms responsible for these characteristics are not well understood. Adaptive changes in mRNA expression in equine skeletal muscle were investigated by real-time qRT-PCR for a panel of candidate exercise-response genes following a standardized incremental-step treadmill exercise test in eight untrained Thoroughbred horses. Biopsy samples were obtained from the gluteus medius before, immediately after, and 4 h after exercise. Significant (P < 0.05) differences in gene expression were detected for six genes (CKM, COX4I1, COX4I2, PDK4, PPARGC1A, and SLC2A4) 4 h after exercise. Investigation of relationships between mRNA and velocity at maximum heart rate (VHR(max)) and peak postexercise plasma lactate concentration ([La]T(1)) revealed significant (P < 0.05) associations with postexercise COX4I1 and PPARCG1A expression and between [La]T(1) and basal COX4I1 expression. Gene expression changes were investigated in a second cohort of horses after a 10 mo period of training. In resting samples, COX4I1 gene expression had significantly increased following training, and, after exercise, significant differences were identified for COX4I2, PDK4, and PPARGC1A. Significant relationships with VHR(max) and [La]T(1) were detected for PPARGC1A and COX4I1. These data highlight the roles of genes responsible for the regulation of oxygen-dependent metabolism, glucose metabolism, and fatty acid utilization in equine skeletal muscle adaptation to exercise.

A comprehensive characterisation of the fibre composition and properties of a limb (flexor digitorum superficialis, membri thoraci) and a trunk (psoas major) muscle in cattle.

BACKGROUND: The fibre type attributes and the relationships among their properties play an important role in the differences in muscle capabilities and features. Comprehensive characterisation of the skeletal muscles should study the degree of association between them and their involvement in muscle functionality. The purposes of the present study were to characterise the fibre type composition of a trunk (Psoas major, PM) and a limb (Flexor digitorum, membri thoraci, FD) muscle in the bovine species and to study the degree of coordination among contractile, metabolic and histological properties of fibre types. Immunohistochemical, histochemical and histological techniques were used. RESULTS: The fibre type composition was delineated immunohistochemically in calf muscle samples, identifying three pure (I, IIA, and IIX) and two hybrid type fibres (I+IIA, and IIAX). Most of the fibres in FD were types I and IIA, while pure IIX were absent. All fibre types were found in PM, the IIX type being the most frequent. Compared to other species, small populations of hybrid fibres were detected. The five fibre types, previously identified, were ascribed to three different acid and alkaline mATPase activity patterns. Type I fibres had the highest oxidative capacity and the lowest glycolytic capacity. The reverse was true for the IIX fibres, whereas the type IIA fibres showed intermediate properties. Regarding the histological properties, type I fibres tended to be more capillarised than the II types. Correlations among contractile, metabolic and histological features on individual fibres were significantly different from zero (r values varied between -0.31 and 0.78). Hybrid fibre values were positioned between their corresponding pure types, and their positions were different regarding their metabolic and contractile properties. CONCLUSION: Coordination among the contractile, metabolic and histological properties of fibres has been observed. However, the magnitude of the correlation among them is always below 0.8, suggesting that the properties of muscles are not fully explained by the fibre composition. These results support the concept that, to some extent, muscle plasticity can be explained by the fibre type composition, and by the properties derived from their metabolic and histological profiles.

Slow- and fast-twitch rat hind limb skeletal muscle phenotypes 8 months after spinal cord transection and olfactory ensheathing glia transplantation.

Paralysed skeletal muscle of rats with spinal cord injury (SCI) undergoes atrophy and a switch in gene expression pattern which leads to faster, more fatigable phenotypes. Olfactory ensheathing glia (OEG) transplants have been reported to promote axonal regeneration and to restore sensory-motor function in animals with SCI. We hypothesized that OEG transplants could attenuate skeletal muscle phenotypic deterioration and that this effect could underlie the functional recovery observed in behavioural tests. A variety of morphological, metabolic and molecular markers were assessed in soleus (SOL) and extensor digitorum longus (EDL) muscles of spinal cord transected (SCT), OEG-transplanted rats 8 months after the intervention and compared with non-transplanted SCT rats and sham-operated (without SCT) controls (C). A multivariate analysis encompassing all the parameters indicated that OEG-transplanted rats displayed skeletal muscle phenotypes intermediate between non-transplanted and sham-operated controls, but different from both. A high correlation was observed between behaviourally tested sensory-motor functional capacity and expression level of slow- and fast-twitch hind limb skeletal muscle phenotypic markers, particularly the histochemical glycerol-3-phosphate dehydrogenase activity (-0.843, P < 0.0001) and the fraction of variant 2s of the slow regulatory myosin light chain isoform (0.848, P < 0.0001) in SOL. Despite the mean overall effect of OEG transplants in patterning skeletal muscle protein expression towards normal, in 6 out of 9 animals they appeared insufficient to overcome fibre type switching and to support a consistent and generalized long-term maintenance of normal skeletal muscle characteristics. The interplay of OEG and exercise-mediated neurotrophic actions is a plausible mechanism underlying OEG transplantation effects on paralysed skeletal muscle.

Effects of intensity and duration of exercise on muscular responses to training of thoroughbred racehorses.

This study examined the effects of the intensity and duration of exercise on the nature and magnitude of training adaptations in muscle of adolescent (2-3 yr old) racehorses. Six thoroughbreds that had been pretrained for 2 mo performed six consecutive conditioning programs of varying lactate-guided intensities [velocities eliciting blood lactate concentrations of 2.5 mmol/l (v2.5) and 4 mmol/l (v4), respectively] and durations (5, 15, 25 min). Pre- and posttraining gluteus muscle biopsies were analyzed for myosin heavy chain content, fiber-type composition, fiber size, capillarization, and fiber histochemical oxidative and glycolytic capabilities. Although training adaptations were similar in nature, they varied greatly in magnitude among the different training protocols. Overall, the use of v4 as the exercise intensity for 25 min elicited the most consistent training adaptations in muscle, whereas the minimal training stimulus that evoked any significant change was identified with exercises of 15 min at v2.5. Within this range, muscular adaptations showed significant trends to be proportional to the exercise load of specific training programs. Taken together, these data suggest that muscular adaptations to training in horses occur on a continuum that is based on the exercise intensity and duration of training. The practical implications of this study are that exercises for 15 to 25 min/day at velocities between v2.5 and v4 can improve in the short term (3 wk) the muscular stamina in thoroughbreds. However, exercises of 5-15 min at v4 are necessary to enhance muscular features related to strength (hypertrophy).

New insights into skeletal muscle fibre types in the dog with particular focus towards hybrid myosin phenotypes.

Electrophoresis, immunoblots, immunohistochemistry and image analysis methods were applied to characterise canine trunk and appendicular muscle fibres according to their myosin heavy chain (MyHC) composition and to determine, on a fibre-to-fibre basis, the correlation between contractile [MyHC (s), myofibrillar ATPase (mATPase) and sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) isoforms], metabolic [succinate dehydrogenase (SDH) and glycerol-3-phosphate dehydrogenase (GPDH) activities and glycogen and phospholamban (PLB) content] and morphological (cross-sectional area and capillary and nuclear densities) features of individual myofibres. An accurate delineation of MyHC-based fibre types was obtained with the developed immunohistochemical method, which showed high sensitivity and objectivity to delineate hybrid fibres with overwhelming dominance of one MyHC isoform. Phenotypic differences in contractile, metabolic and morphological properties seen between fibre types were related to MyHC content. All canine skeletal muscle fibre types had a relatively high histochemical SDH activity but significant differences existed in the order IIA>I>IIX. Mean GPDH was ranked according to fibre type such that II>IIX. Hybrid fibres, which represented nearly one third of the whole pool of skeletal muscle fibres analysed, had mean values intermediate between their respective pure phenotypes. Slow fibres expressed the slow SERCA isoform and PLB, whereas type II fibres expressed the fast SERCA isoform. Discrimination of myofibres according to their MyHC content was possible on the basis of their contractile, metabolic and morphological features. These intrafibre interrelationships suggest that myofibres of control dogs exhibit a high degree of co-ordination in their physiological, biochemical and morphological characteristics. This study demonstrates that canine skeletal muscle fibres have been misclassified in numerous previous studies and offers useful baseline data and new prospects for future work on muscle-fibre-typing in canine experimental studies.

Myosin heavy chain fibre types and fibre sizes in nuliparous and primiparous ovariectomized Iberian sows: Interaction with two alternative rearing systems during the fattening period.

To investigate the effects of the reproductive background (nuliparous/primiparous) on skeletal muscle phenotype and their interaction with two alternative finishing systems (indoors/outdoors) in Iberian sows, 32 contemporary piglets were assigned to four different groups: indoors-nuliparous, indoors-primiparous, outdoors-nuliparous and outdoors-primiparous. Nuliparous sows were ovariectomized at 4 months of age, whereas primiparous were castrated at 14 months of age, after their first lactation. Indoors sows were fattened (2 months prior to slaughter) intensively, whereas outdoors sows were finished in an extensive land with natural resources. Muscle samples (semimembranosus, gluteobiceps) were analyzed for myosin fibre types and fibre sizes. Sows finished out-of-doors had more type I and IIA fibres and smaller fibres than indoor-fattened pigs. Primiparous reared indoors had a significantly lower IIX:IIB fibre ratio and showed a clear tendency for having larger fibres than nuliparous reared indoors. Reproductive background effect interacted with rearing condition effect inducing relevant muscular adaptations. Thus, the influence of reproductive background on muscle characteristics was not significant in sows reared out-of-doors but highly significant in sows finished intensively. This interaction resulted in muscular traits less favourable for meat quality in primiparous vs. nuliparous reared indoors.

New insights into the skeletal muscle phenotype of equine motor neuron disease: a quantitative approach.

Equine motor neuron disease (EMND) is a neurodegenerative disorder similar to the sporadic form of human amyotrophic lateral sclerosis. This study was conducted to quantify myofiber plasticity in response to EMND. Deep M. gluteus medius biopsy samples from eight horses with an ante mortem diagnosis of EMND, which in five cases was later confirmed by post mortem examination of spinal cord and peripheral nerves, were examined by combined methodologies of electrophoresis of myosin heavy chains (MyHC), muscle enzymes and substrate biochemistry, immunohistochemistry of MyHCs and sarcoendoplasmic Ca2+-ATPase (SERCA) isoforms, quantitative histochemistry of succinic dehydrogenase, glycerol-3-phosphate dehydrogenase, periodic acid-Schiff and capillaries, and photometric image analysis. The data were compared with muscle biopsies from healthy controls. Histopathological findings of EMND were observed in muscle biopsy specimens from all cases, but the severity and intra-biopsy extent varied from case to case. Compared with controls, muscle biopsy samples from EMND horses had a lower percentage of MyHC type I fibers, higher percentages of hybrid IIAX and pure IIX fibers, significant atrophy of all muscle fiber types, reduced oxidative capacity, increased glycolytic capacity, diminished intramuscular glycogen, lower capillary-to-fiber ratio, a higher ratio of myofibers expressing SERCA1a to SERCA2a isoforms, and a lower percentage of fibers expressing phospholamban. Objective discrimination of muscle biopsy specimens according to their healthy status (EMND vs controls) was possible on the basis of their muscular characteristics. A coordinated shift from slow to fast muscle characteristics in contractile and metabolic features of muscle fiber types, together with generalized myofiber atrophy, occurs in EMND and the extent of this change seems to be related to the duration of the disease.

Coordinated expression of myosin heavy chains, metabolic enzymes, and morphological features of porcine skeletal muscle fiber types.

Combined methodologies of electrophoresis, immunoblots, immunohistochemistry, histochemistry, and photometric image analysis were applied to characterize porcine skeletal muscle fibers according to their myosin heavy chain (MyHC) composition, and to determine on a fiber-to-fiber basis the correlation between contractile [MyHC (s), myofibrillar ATPase (mATPase), and sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) isoforms], metabolic [succinate dehydrogenase (SDH), and glycerol-3-phosphate dehydrogenase (GPDH) activities, glycogen, and phospholamban (PLB) contents], and morphological [cross-sectional area (CSA), capillary, and nuclear densities] features of individual myofibers. An accurate delineation of MyHC-based fiber types was obtained with the immunohistochemical method developed. This protocol showed a high sensitivity and objectivity to delineate hybrid fibers with overwhelming dominance of one MyHC isoform. The phenotypic differences in contractile, metabolic, and morphological properties seen between fiber types were related with MyHC content. Slow fibers had the lowest mATPase activity (related to shortening velocity), the highest SDH activity (oxidative capacity), the lowest GPDH activity (glycolytic metabolism), and glycogen content, the smallest CSA, the greatest capillary, and nuclear densities, and expressed slow SERCA isoform and PLB, but not the fast SERCA isoform. The reverse pattern was true for pure IIB fibers, whereas type IIA and IIX fibers had intermediate properties. Hybrid fibers had mean values intermediate in-between their respective pure phenotypes. Discrimination of myofibers according to their MyHC content was possible on the basis of their contractile and non-contractile profiles. These intrafiber interrelationships suggest that myofibers of control pigs exhibit a high degree of co-ordination in their physiological, biochemical, and anatomical features. This study may well be a useful baseline for future work on the pig meat industry and also offers new prospects for muscle fiber typing in porcine experimental studies.

Neuromuscular partitioning, architectural design, and myosin fiber types of the M. vastus lateralis of the llama (Lama glama).

The llama (Lama glama) is one of the few mammals of relatively large body size in which three fast myosin heavy chain isoforms (i.e., IIA, IIX, IIB) are extensively expressed in their locomotory muscles. This study was designed to gain insight into the morphological and functional organization of skeletal musculature in this peculiar animal model. The neuromuscular partitioning, architectural design, and myosin fiber types were systematically studied in the M. vastus lateralis of adult llamas (n = 15). Four nonoverlapping neuromuscular partitions or compartments were identified macroscopically (using a modified Sihler's technique for muscle depigmentation), although they did not conform strictly to the definitions of "neuromuscular compartments." Each neuromuscular partition was innervated by primary branches of the femoral nerve and was arranged within the muscle as paired partitions, two in parallel (deep-superficial compartmentalization) and the other two in-series (proximo-distal compartmentalization). These neuromuscular partitions of the muscle varied in their respective architectural designs (studied after partial digestion with diluted nitric acid) and myosin fiber type characteristics (identified immunohistochemically with specific anti-myosin monoclonal antibodies, then examined by quantitative histochemistry and image analysis). The deep partitions of the muscle had longer fibers, with lower angles of pinnation, and higher percentages of fast-glycolytic fibers than the superficial partitions of the muscle. These differences clearly suggest a division of labor in the whole M. vastus lateralis of llamas, with deep partitions exhibiting features well adapted for dynamic activities in the extension of stifle, whereas superficial portions seem to be related to the antigravitational role of the muscle in preserving the extension of the stifle during standing and stance phase of the stride. This peculiar structural and functional organization of the llama M. vastus lateralis does not confirm the generalized idea that deep muscles or the deepest portions within the same muscles somehow develop postural and/or low-intensity isometric functions. Rather, it suggests a primacy of architecture over intramuscular location in determining fiber type composition and hence division of labor within the muscle. A compartmentalization in the distribution of the three fast-subtype fibers (IIA, IIX, and IIB) also occurred, and this could also be relevant functionally, since these fiber types differed significantly in size (IIA < IIX < IIB), oxidative capacity (IIA > IIX > IIB), and capillarization (IIA = IIX > IIB). Furthermore, a typical spatial pattern in fiber type distribution was encountered in llama muscle (i.e., fiber types were consistently ranked in the order I --> IIA --> IIX --> IIB from the center to the periphery of fascicles), suggesting again peculiar and not well understood functional adaptations in these species.

Determinants of VO(2) kinetics at high power outputs during a ramp exercise protocol.

PURPOSE: To determine the relationship between the additional, nonlinear increase in oxygen uptake (Delta VO(2)) that occurs at high power outputs during a ramp cycle ergometer test, on one hand; and possible explanatory mechanisms of the phenomenon, such as cardiorespiratory work, blood lactate, fitness level, or muscle fiber distribution, on the other. METHODS: Ten healthy, sedentary young adults (age (mean +/- SEM), 22 +/- 1 yr) were chosen as subjects. A muscle biopsy specimen was taken from the vastus lateralis of the right leg to determine fiber type distribution by immunohistochemical identification of myosin heavy chain (MHC) isoforms. During the ramp tests (power output increases of 5 W every 15-s interval), the ventilatory threshold (VT) and lactate threshold (LT) were measured. We defined Delta VO(2) as the difference between "true" VO(2) values observed at the maximal power output (VO(2)obs) and those expected (VO(2)exp) from the previous linear VO2:power output relationship below the VT. RESULTS: A nonlinear increase was observed in VO2 (Delta VO(2) = 239 +/- 79 mL x min(-1), P < 0.05 for VO(2)obs vs VO(2)exp), which was significantly correlated with the percentage of type IIX fibers (r = 0.80, P < 0.05). No other correlations were found between Delta VO(2) and possible explanatory mechanisms. CONCLUSION: A greater percentage of type IIX fibers is associated with a higher excess VO(2) at high power outputs (above VT).