Effects of marketable ages on meat quality through fiber characteristics in the goose.

Goose meat is increasingly popular among consumers because of its good quality. The fiber characteristics have been well demonstrated to be key contributing factors of meat quality, and the marketable ages are also closely related to meat quality. However, little is known about the effect of different marketable ages on the quality of goose meat through its fiber characteristics. Here, fiber characteristics of Yangzhou geese of different marketable ages (70, 90, and 120 d) and their effect on meat quality were investigated. The results showed that only fast-twitch fibers were present in breast muscle, irrespective of age, and that few slow-twitch fibers could be identified in leg muscle, especially in gastrocnemius and extensor digitorum longus. Fiber diameter in breast muscle increased rapidly from age 70 d to 90 d, from 19.88 to 26.27 µm, and remained stable for 90 d thereafter. The diameter and cross-sectional area of muscle fiber continue to grow with day increasing in leg muscle. In addition, we measured the proximate composition and physical properties at different ages. Among the 3 marketable ages investigated, the 120-day-old geese had higher intramuscular fat and protein content, as well as lower moisture content, both in breast and leg meat. Greater lightness and pressing loss, with lower redness and shear force, were observed in the breast and leg meat of 70-day-old geese when compared with 90- or 120-day-old geese. Taken together, although older marketable age hardly affected muscle fiber type in geese, it would contribute to larger muscle fiber area, higher intramuscular fat and protein content, as well as redder and chewier meat. As a result, the reasonable marketable age should be taken into account to improve quality in goose meat production, and the marketable age of 90 or 120 d was recommended and it could potentially improve meat quality in goose meat production.

Influence of sex and fiber type on the satellite cell pool in human skeletal muscle.

The repair, remodeling, and regeneration of myofibers are dependent on satellite cells (SCs), although, the distribution of SCs in different fiber types of human muscle remains inconclusive. There is also a paucity of research comparing muscle fiber characteristics in a sex-specific manner. Therefore, the aim of this study was to investigate fiber type-specific SC content in men and women. Muscle biopsies from vastus lateralis were collected from 64 young (mean age 27 ± 5), moderately trained men (n = 34) and women (n = 30). SCs were identified by Pax7-staining together with immunofluorescent analyses of fiber type composition, fiber size, and myonuclei content. In a mixed population, comparable number of SCs was associated to type I and type II fibers (0.07 ± 0.02 vs 0.07 ± 0.02 SCs per fiber, respectively). However, unlike men, women displayed a fiber type-specific distribution, with SC content being lower in type II than type I fibers (P = .041). Sex-based differences were found specifically for type II fibers, where women displayed lower SC content compared to men (P < .001). In addition, positive correlations (r-values between 0.36-0.56) were found between SC content and type I and type II fiber size in men (P = .03 and P < .01, respectively), whereas similar relationships could not be detected in women. Sex-based differences were also noted for fiber type composition and fiber size, but not for myonuclei content. We hereby provide evidence for sex-based differences present at the myocellular level, which may have important implications when studying exercise- and training-induced myogenic responses in skeletal muscle.

Tipo de fibra muscular y su relación con el abordaje fonoaudiológico en los trastornos de la deglución / Type of muscle fibreand its relationship to the Speech-Language Pathology approach in swallowing disorders

Los actuales protocolos de intervención en deglución no especifican parámetros de los ejercicios tales como la cantidad de series, duración, tiempos de repetición, entre otros. Lo anterior, es un desafío para la intervención fonoaudiológica en los trastornos de la deglución. El objetivo de este trabajo es acercar al profesional fonoaudiólogo al conocimiento de las fibras musculares, ya que es un conocimiento que se debe considerar antes de indicar ejercicios musculares cráneo-cérvico-orales. Por ello, se describen las principales fibras musculares con sus respectivas características fundamentales, como lo son: la resistencia a la fatiga y la velocidad de contracción. Se revisa la literatura sobre la dispersión de las fibras musculares, de algunos de los principales músculos que participan en el proceso deglutorio. También, se analizan diversas particularidades de los músculos de la zona cráneo-cérvico-oral. Se describen además las diferentes dificultades para evaluar esta musculatura. Finalmente, se expone la relevancia práctica de conocer estos tipos de fibras musculares y las perspectivas futuras de este enfoque basado en parámetros del ejercicio y la medición de variables objetivas.

Swallowing intervention protocols in dysphagia do not specify the parameters of the exercises, namely the duration, frequencyand number of series, all of which present a challenge in speech therapy intervention in swallowing disorders. The purpose of this review is to provide Speech and Language Pathologists with background information concerning crucial muscle fibres used in swallowing therapy and to subsequently indicate appropriate skull-cervical-oral muscle exercises. We describe the primary muscle and its characteristics such as fatigue resistance and contraction speed, review the types of fibres of some of the main muscles that participate in the swallowing process, and present the particularities of the muscles of the cranio-cervical-oral area in comparison with the rest of the skeletal muscles. We also provide information regarding the difficulties in evaluating the musculature involvedin the swallowing process before finally highlighting the relevance of understanding the roles and characteristics of these muscles for clinical practice

PPARGC1A rs8192678 and NRF1 rs6949152 Polymorphisms Are Associated with Muscle Fiber Composition in Women.

PPARGC1A rs8192678 G/A (Gly482Ser) and NRF1 rs6949152 A/G polymorphisms have been associated with endurance athlete status, endurance performance phenotypes, and certain health-related markers of different pathologies such as metabolic syndrome, diabetes, and dyslipidemia. We hypothesized that they could be considered interesting candidates for explaining inter-individual variations in muscle fiber composition in humans. We aimed to examine possible associations of these polymorphisms with myosin heavy-chain (MHC) isoforms as markers of muscle fiber compositions in vastus lateralis muscle in a population of 214 healthy Japanese subjects, aged between 19 and 79 years. No significant associations were found in men for any measured variables. In contrast, in women, the PPARGC1A rs8192678 A/A genotype was significantly associated with a higher proportion of MHC-I (p = 0.042) and with a lower proportion of MHC-IIx (p = 0.033), and the NRF1 rs6949152 AA genotype was significantly associated with a higher proportion of MHC-I (p = 0.008) and with a lower proportion of MHC IIx (p = 0.035). In women, the genotype scores of the modes presenting the most significant results for PPARGC1A rs8192678 G/A (Gly482Ser) and NRF1 rs6949152 A/G polymorphisms were significantly associated with MHC-I (p = 0.0007) and MHC IIx (p = 0.0016). That is, women with combined PPARGC1A A/A and NRF1 A/A genotypes presented the highest proportion of MHC-I and the lowest proportion of MHC-IIx, in contrast to women with combined PPARGC1A GG+GA and NRF1 AG+GG genotypes, who presented the lowest proportion of MHC-I and the highest proportion of MHC-IIx. Our results suggest possible associations between these polymorphisms (both individually and in combination) and the inter-individual variability observed in muscle fiber composition in women, but not in men.

Exploring the lncRNAs Related to Skeletal Muscle Fiber Types and Meat Quality Traits in Pigs.

The alteration in skeletal muscle fiber is a critical factor affecting livestock meat quality traits and human metabolic diseases. Long non-coding RNAs (lncRNAs) are a diverse class of non-coding RNAs with a length of more than 200 nucleotides. However, the mechanisms underlying the regulation of lncRNAs in skeletal muscle fibers remain elusive. To understand the genetic basis of lncRNA-regulated skeletal muscle fiber development, we performed a transcriptome analysis to identify the key lncRNAs affecting skeletal muscle fiber and meat quality traits on a pig model. We generated the lncRNA expression profiles of fast-twitch Biceps femoris (Bf) and slow-twitch Soleus (Sol) muscles and identified the differentially expressed (DE) lncRNAs using RNA-seq and performed bioinformatics analyses. This allowed us to identify 4581 lncRNA genes among six RNA libraries and 92 DE lncRNAs between Bf and Sol which are the key candidates for the conversion of skeletal muscle fiber types. Moreover, we detected the expression patterns of lncRNA MSTRG.42019 in different tissues and skeletal muscles of various development stages. In addition, we performed a correlation analyses between the expression of DE lncRNA MSTRG.42019 and meat quality traits. Notably, we found that DE lncRNA MSTRG.42019 was highly expressed in skeletal muscle and its expression was significantly higher in Sol than in Bf, with a positive correlation with the expression of Myosin heavy chain 7 (MYH7) (r = 0.6597, p = 0.0016) and a negative correlation with meat quality traits glycolytic potential (r = -0.5447, p = 0.0130), as well as drip loss (r = -0.5085, p = 0.0221). Moreover, we constructed the lncRNA MSTRG.42019-mRNAs regulatory network for a better understanding of a possible mechanism regulating skeletal muscle fiber formation. Our data provide the groundwork for studying the lncRNA regulatory mechanisms of skeletal muscle fiber conversion, and given the importance of skeletal muscle fiber types in muscle-related diseases, our data may provide insight into the treatment of muscular diseases in humans.

A proposal for a new classification of soleus muscle morphology.

INTRODUCTION: The soleus muscle (SM) is located in the superficial posterior compartment of the leg, together with the gastrocnemius muscle (GM) and plantaris muscle. There is little information on the morphological variability (pennation) of the SM. The aim of the study is to characterize the variations in the morphology of the pennation of the SM and to create the first accurate classification. MATERIAL AND METHODS: Eighty lower limbs (40 left, and 40 right) fixed in 10% formalin solution were examined. The morphology of the central tendon, medial and lateral aponeuroses was evaluated, together with the pennation angle. RESULTS: In all cases, the soleus muscle was present. Four types could be distinguished based on muscle fibre morphology. In order of frequency: Type 1 - bipennate (43.75%); Type 4 - non-pennate (36.25%); Type 2 - unipennate (15%); Type 3 - multipennate (5%). No significant difference in type distribution was observed with regard to body side (p=0.9018) or gender (p=0.0844). CONCLUSION: The soleus muscle is characterized by high morphological variability. Based on the pennation angle, four types (1-4) of soleus muscle are distinguishable.

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples.

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.

A data-driven methodology reveals novel myofiber clusters in older human muscles.

Skeletal muscles control posture, mobility and strength, and influence whole-body metabolism. Muscles are built of different types of myofibers, each having specific metabolic, molecular, and contractile properties. Fiber classification is, therefore, regarded the key for understanding muscle biology, (patho-) physiology. The expression of three myosin heavy chain (MyHC) isoforms, MyHC-1, MyHC-2A, and MyHC-2X, marks myofibers in humans. Typically, myofiber classification is performed by an eye-based histological analysis. This classical approach is insufficient to capture complex fiber classes, expressing more than one MyHC-isoform. We, therefore, developed a methodological procedure for high-throughput characterization of myofibers on the basis of multiple isoforms. The mean fluorescence intensity of the three most abundant MyHC isoforms was measured per myofiber in muscle biopsies of 56 healthy elderly adults, and myofiber classes were identified using computational biology tools. Unsupervised clustering revealed the existence of six distinct myofiber clusters. A comparison with the visual assessment of myofibers using the same images showed that some of these myofiber clusters could not be detected or were frequently misclassified. The presence of these six clusters was reinforced by RNA expressions levels of sarcomeric genes. In addition, one of the clusters, expressing all three MyHC isoforms, correlated with histological measures of muscle health. To conclude, this methodological procedure enables deep characterization of the complex muscle heterogeneity. This study opens opportunities to further investigate myofiber composition in comparative studies.

La rigidez cadavérica, el espasmo cadavérico y tipos de fibras musculares / The cadaveric rigidity, the cadaveric spasm and types of muscle fibers

La rigidez cadavérica (rigor mortis) es un proceso no muy bien comprendido por la mayoría de los médicos. El conocimiento de la intimidad del proceso de la rigidez cadavérica es de vital importancia ya que es una de las variables que junto con las livideces (livor mortis) y la temperatura (algor mortis) del cadáver ayudan a determinar el cronotanatodiagnóstico, tanatocronodiagnóstico o intervalo postmortal del período inmediato de la muerte. Para entender el mecanismo de la rigidez y el espasmo cadavérico es preciso hacer un repaso de la contracción muscular fisiológica en el vivo. Hay que tener presente que el tipo de fibra muscular predominante modificará las características de la contracción muscular fisiológica en el vivo, y también la rigidez y el espasmo cadavérico. (AU)

The cadaveric rigidity (rigor mortis) is a process which is not very well understood by the majority of the doctors. The knowledge of the intimacy of the cadaveric stiffness process is of vital importance since it is one of the variables that, as well as the postmortem lividity (livor mortis) and the body temperature post mortem (algor mortis) help determine the chronotanatodiagnostic, tanatochronodiagnostic or postmortal interval of the immediate period of death. In order to understand the mechanism of stiffness and cadaveric spasm, it is necessary to review the physiological muscle contraction in vivo. We should keep in mind that the predominant type of muscle fiber will modify the characteristics of physiological muscle contraction in vivo, as well as stiffness and cadaveric spasm. (AU)

Are there consistent relationships between major connective tissue components, intramuscular fat content and muscle fibre types in cattle muscle?

Intramuscular connective tissue (IMCT) is mainly composed of several fibrils (known as total collagen (TCol)) linked between each other by different chemical cross-links (CLs), the whole being embedded in a matrix of proteoglycans (PGs). In the field of beef quality, there is limited information on the role of CLs and PGs. Accordingly, several authors suggest that, to investigate the role of IMCT, it is important to investigate them just like TCol and insoluble collagen (ICol). In muscle, there are two other components, the muscle fibres and intramuscular fat (IMF) content. There are limited data on the relationships between these three components of muscle and then on possibility to independently manipulate these characteristics in order to control the final quality of meat. The present study aimed to investigate whether consistent relationships exist between these different components of muscle. Therefore, the present study compared four muscles of two cattle types (dairy and beef) to determine associations between TCol, ICol, CLs and PGs. Data were analysed across and within muscle (M) and animal type (AT) based on residuals. There was a strong M and AT effect for all muscle characteristics and an interaction M × AT for type I muscle fibres and IMF. Correlations between TCol, ICol and their CLs were M- and AT-independent. Total proteoglycans were positively correlated with TCol and ICol in a muscle-dependent manner irrespective of AT, but no correlation was found with CLs. On the contrary, CLs were negatively correlated with the ratio TPGs : TCol in an M-dependent manner, irrespective of AT. TCol, ICol and CLs were positively and negatively correlated with type IIA and IIB+X muscle fibres only in longissimus thoracis (LT) muscle, regardless the AT. Insoluble collagen was the only parameter of IMCT to be correlated with type I muscle fibres but only in LT muscle, irrespective of AT. There was no correlation between PGs and muscle fibre types, but PGs were the only IMCT component to be related with IMF in an M-dependent manner, irrespective of AT. Finally, there was no correlation between muscle fibre types and IMF content within M and AT. This study revealed that there is a strong relationship between IMCT components irrespective of M, an M-dependent relationship between the IMCT components and muscle fibre types and few (only with PGs) or no relationship between IMF and IMCT and muscle fibres.

Dietary thymol supplementation promotes skeletal muscle fibre type switch in longissimus dorsi of finishing pigs.

As one of the key points related to meat quality, skeletal muscle fibre type is determined by energy metabolism and genetic factors, but its transformation could be also greatly influenced by many factors. Thymol, the primary effective ingredients of thyme, is well known for its anti-oxidation and anti-inflammatory, while little is known about its effect on skeletal muscle oxidative metabolism and fibre type switch. Therefore, in order to investigate its effects and possibility to be applied in livestock production, 36 150-day-old fattening Pigs were fed with different diet for six-week experiment. As a result, the drip loss ratio of longissimus dorsi (LD) was significantly reduced (p < .05). Oxidative metabolism-related enzyme activity, the mRNA levels and protein expression of COX5B and PGC1α, mRNA level of myosin heavy chain I (MyHC I) and protein level of MyHC IIa were significantly upregulated (p < .05). While compared with control group, the protein expression of MyHC IIb was significantly decreased (p < .05). The result revealed that thymol could promote the oxidative metabolism in the muscle of pigs and improve the meat quality to a certain extent.

Automated image-analysis method for the quantification of fiber morphometry and fiber type population in human skeletal muscle.

BACKGROUND: The quantitative analysis of muscle histomorphometry has been growing in importance in both research and clinical settings. Accurate and stringent assessment of myofibers' changes in size and number, and alterations in the proportion of oxidative (type I) and glycolytic (type II) fibers is essential for the appropriate study of aging and pathological muscle, as well as for diagnosis and follow-up of muscle diseases. Manual and semi-automated methods to assess muscle morphometry in sections are time-consuming, limited to a small field of analysis, and susceptible to bias, while most automated methods have been only tested in rodent muscle. METHODS: We developed a new macro script for Fiji-ImageJ to automatically assess human fiber morphometry in digital images of the entire muscle. We tested the functionality of our method in deltoid muscle biopsies from a heterogeneous population of subjects with histologically normal muscle (male, female, old, young, lean, obese) and patients with dermatomyositis, necrotizing autoimmune myopathy, and anti-synthetase syndrome myopathy. RESULTS: Our macro is fully automated, requires no user intervention, and demonstrated improved fiber segmentation by running a series of image pre-processing steps before the analysis. Likewise, our tool showed high accuracy, as compared with manual methods, for identifying the total number of fibers (r = 0.97, p < 0.001), fiber I and fiber II proportion (r = 0.92, p < 0.001), and minor diameter (r = 0.86, p < 0.001) while conducting analysis in ~ 5 min/sample. The performance of the macro analysis was maintained in pectoral and deltoid samples from subjects of different age, gender, body weight, and muscle status. The output of the analyses includes excel files with the quantification of fibers' morphometry and color-coded maps based on the fiber's size, which proved to be an advantageous feature for the fast and easy visual identification of location-specific atrophy and a potential tool for medical diagnosis. CONCLUSION: Our macro is reliable and suitable for the study of human skeletal muscle for research and for diagnosis in clinical settings providing reproducible and consistent analysis when the time is of the utmost importance.

Effect of skeletal muscle phenotype and gender on fasting-induced myokine expression in mice.

Skeletal muscle secretes myokines, which are involved in metabolism and muscle function regulation. The role of fasting on myokine expression in skeletal muscle is largely unknown. In this study, we used gastrocnemius skeletal muscle RNA sequencing data from fasting male mice in the Gene Expression Omnibus (GEO) database. Adopted male and female C57BL/6J mice that fasted for 24 h were included to examine the effect of fasting on myokine expression in slow-twitch soleus and fast-twitch tiabialis anterior (TA) skeletal muscle. We found that fasting significantly affected many myokines in muscle. Fasting reduced Fndc5 and Igf1 gene expression in soleus and TA muscles in both male and female mice without muscle phenotype or gender differences, but Il6, Mstn and Erfe expression was influenced by fasting with fibre type- and gender-dependent effects. Fasting also induced muscle atrophy marker genes Murf1 and Fbxo32 and reduced myogenesis factor Mef2 expression without muscle fibre or gender differences. We further found that the expression of transcription factors Pgc1α, Pparα, Pparγ and Pparδ had muscle fibre type-dependent effects, and the expression of Pgc1α and Pparα had gender-dependent effects. The sophisticated expression pattern of myokines would partially explain the complicated cross-talk between skeletal muscle and other organs in different genders and muscles phenotypes, and it is worth further investigation.

A fast, reliable and sample-sparing method to identify fibre types of single muscle fibres.

Many skeletal muscle proteins are present in a cell-specific or fibre-type dependent manner. Stimuli such as exercise, aging, and disease have been reported to result in fibre-specific responses in protein abundances. Thus, fibre-type-specific determination of the content of specific proteins provides enhanced mechanistic understanding of muscle physiology and biochemistry compared with typically performed whole-muscle homogenate analyses. This analysis, however, is laborious and typically not performed. We present a novel dot blotting method for easy and rapid determination of skeletal muscle fibre type based on myosin heavy chain (MHC) isoform presence. Requiring only small amounts of starting muscle tissue (i.e., 2-10 mg wet weight), muscle fibre type is determined in one-tenth of a 1-3-mm fibre segment, with the remainder of each segment pooled with fibre segments of the same type (I or II) for subsequent protein quantification by western blotting. This method, which we validated using standard western blotting, is much simpler and cheaper than previous methods and is adaptable for laboratories routinely performing biochemical analyses. Use of dot blotting for fibre typing will facilitate investigations of fibre-specific responses to diverse stimuli, which will advance our understanding of skeletal muscle physiology and biochemistry.

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.

High-throughput data-driven analysis of myofiber composition reveals muscle-specific disease and age-associated patterns.

Contractile properties of myofibers are dictated by the abundance of myosin heavy chain (MyHC) isoforms. MyHC composition designates muscle function, and its alterations could unravel differential muscle involvement in muscular dystrophies and aging. Current analyses are limited to visual assessments in which myofibers expressing multiple MyHC isoforms are prone to misclassification. As a result, complex patterns and subtle alterations are unidentified. We developed a high-throughput, data-driven myofiber analysis to quantitatively describe the variations in myofibers across the muscle. We investigated alterations in myofiber composition between genotypes, 2 muscles, and 2 age groups. We show that this analysis facilitates the discovery of complex myofiber compositions and its dependency on age, muscle type, and genetic conditions.-Raz, V., Raz, Y., van de Vijver, D., Bindellini, D., van Putten, M., van den Akker, E. B. High-throughput data-driven analysis of myofiber composition reveals muscle-specific disease and age-associated patterns.

Muscle fiber type diversity revealed by anti-myosin heavy chain antibodies.

Different forms of myosin heavy chains (MyHCs), coded by a large family of sarcomeric MYH genes, are expressed in striated muscles. The generation of specific anti-MyHC antibodies has provided a powerful tool to define the fiber types present in skeletal muscles, their functional properties, their response to conditions that affect muscle plasticity and their changes in muscle disorders. Cardiomyocyte heterogeneity has been revealed by the serendipitous observation that different MyHCs are present in atrial and ventricular myocardium and in heart conduction tissue. Developmental MyHCs present in embryonic and fetal/neonatal skeletal muscle are re-expressed during muscle regeneration and can be used to identify regenerating fibers in muscle diseases. MyHC isoforms provide cell type-specific markers to identify the signaling pathways that control muscle cell identity and are an essential reference to interpret the results of single-cell transcriptomics and proteomics.

Promoter analysis of the fish gene of slow/cardiac-type myosin heavy chain implicated in specification of muscle fiber types.

Vertebrate skeletal muscles consist of heterogeneous tissues containing various types of muscle fibers, where specification of the fiber type is crucial for muscle development. Fish are an attractive experimental model to study the mechanisms of such fiber type specification because of the separated localization of slow and fast muscles in the trunk myotome. We examined regulation of expression of the torafugu gene of slow/cardiac-type myosin heavy chain, MYH M5 , and isolated an operational promoter in order to force its tissue-specific expression across different fish species via the transgenic approach in zebrafish and medaka. This promoter activity was observed in adaxial cell-derived superficial slow muscle fibers under the control of a hedgehog signal. We also uncovered coordinated expression of MYH M5 and Sox6b, which is an important transcriptional repressor for specification of muscle fiber types and participates in hedgehog signaling. Sequence comparison in the 5'-flanking region identified three conserved regions, CSR1-CSR3, between torafugu MYH M5 and its zebrafish ortholog. Analysis of deletion mutants showed that CSR1 significantly stimulates gene expression in slow muscle fibers. In contrast, deletion of CSR3 resulted in ectopic expression of a reporter gene in fast muscle fibers. CSR3 was found to contain a putative Sox family protein-binding site. These results indicate that the dual mechanism causing inhibition in fast muscle fibers and activation in slow muscle fibers is essential for slow muscle fiber-specific gene expression in fish.

Skeletal Muscle Toolbox.

Evaluation of skeletal muscle frequently combines morphologic and morphometric techniques. As is the case with many organ systems, skeletal muscle has limited responses to insult or injury. Over the past several years, crucial interactions between skeletal muscle, bone, and the nervous system have been described. The aim of this lecture was to give attendees the necessary background information in basic skeletal muscle morphology, important species differences, introduction to skeletal muscle biomarkers, approaches to morphologic and morphometric evaluation, and examples of background findings and typical responses of skeletal muscle to insult or injury.

High-fat diet induces skeletal muscle oxidative stress in a fiber type-dependent manner in rats.

This study investigated the effects of high-fat (HF) diet on parameters of oxidative stress among muscles with distinct fiber type composition and oxidative capacities. To accomplish that, male Wistar rats were fed either a low-fat standard chow (SC) or a HF diet for 8 weeks. Soleus, extensor digitorum longus (EDL), and epitrochlearis muscles were collected and mitochondrial H2O2 (mtH2O2) emission, palmitate oxidation, and gene expression and antioxidant system were measured. Chronic HF feeding enhanced fat oxidation in oxidative and glycolytic muscles. It also caused a significant reduction in mtH2O2 emission in the EDL muscle, although a tendency towards a reduction was also found in the soleus and epitrochlearis muscles. In the epitrochlearis, HF diet increased mRNA expression of the NADPH oxidase complex; however, this muscle also showed an increase in the expression of antioxidant proteins, suggesting a higher capacity to generate and buffer ROS. The soleus muscle, despite being highly oxidative, elicited H2O2 emission rates equivalent to only 20% and 35% of the values obtained for EDL and epitrochlearis muscles, respectively. Furthermore, the Epi muscle with the lowest oxidative capacity was the second highest in H2O2 emission. In conclusion, it appears that intrinsic differences related to the distribution of type I and type II fibers, rather than oxidative capacity, drove the activity of the anti- and pro-oxidant systems and determine ROS production in different skeletal muscles. This also suggests that the impact of potentially deleterious effects of ROS production on skeletal muscle metabolism/function under lipotoxic conditions is fiber type-specific.