Correlation between histochemically assessed fiber type distribution and isomyosin and myosin heavy chain content in porcine skeletal muscles.

Highly sensitive enzyme assays developed to differentiate skeletal muscle fibers allow the recognition of three main fiber types: slow-twitch oxidative (SO), fast-twitch oxidative glycolytic (FOG), and fast-twitch glycolytic (FG). Myosin, the predominant contractile protein in mammalian skeletal muscle, can be separated based on the electrophoretic mobility under nondissociating conditions into SM2, SM1, IM, FM3, and FM2 isoforms, or under dissociating conditions into myosin heavy chain (MHC) I, IIb, IIx/d, and IIa. The purpose of the present study was to determine whether the histochemical method of differentiation of fiber types is consistent with the electrophoretically identified isomyosin and MHC isoforms. These comparisons were made using serratus ventralis (SV), gluteus medius (GM), and longissimus muscles (LM) from 13 pigs. Two calculation methods for the histochemical assessed fiber type distribution were adopted. The first method incorporated the number of fibers counted for each fiber type and calculated a percentage of the total fiber number (fiber number percentage: FNP). The second method expressed the cross-sectional area of each fiber type as a percentage of the total fiber area measured per muscle (fiber area percentage: FAP). Independent of the calculation methods, correlation analyses revealed in all muscles a strong relation between SO fibers, the slow isomyosin (SM1 and SM2), and MHCI, as well as between the FG fibers, the fast isomyosin (FM3 and FM2), and MHCIIx/b content (P<.05). There were no correlations between FOG fiber population assessed by histochemical analysis and intermediate isoform (IM) or MHCIIa content. The present results did not provide conclusive evidence as to which of the calculation methods (FNP or FAP) was more closely related to myosin composition of skeletal muscles. Despite some incompatibility between the methods, the present study shows that histochemical as well as electrophoretic analyses yielded important information about the composition of porcine skeletal muscle. The combination of the two methods may be essential to accurately characterize porcine skeletal muscles.     

Muscle characteristics and meat quality traits are affected by divergent selection on residual feed intake in pigs

Residual feed intake (RFI) is defined as the difference between the observed feed intake and that expected based on requirements for maintenance and production. A divergent selection was conducted during 4 generations in Large White male pigs to produce low and high RFI lines. The present study aims at determining the influence of this selection on biochemical and histological traits of skeletal muscle, and relating these changes to correlated effects on growth, carcass composition, and meat quality traits. At 8 d preslaughter, biopsies from the LM were taken in the fed state on 14 females from each RFI line fed ad libitum. Animals were slaughtered at 107.8 ± 8.0 kg of BW without any previous fasting. Samples of LM, semimembranosus (SM), biceps femoris (BFM), and rhomboideus muscles were taken at both 30 min and 24 h postmortem. Myofiber typing was only assessed in LM. Low RFI pigs ("efficient") had leaner carcasses with greater muscle content (P < 0.001), less backfat thickness (P < 0.001), and less intramuscular fat content in all 4 muscles (P < 0.01 to P = 0.04). Their greater muscle content was associated with hypertrophy of all fast-twitch fibers. Glycogen content in all glycolytic muscles (i.e., LM, SM and BFM), was greater in low than high RFI pigs. The greater accumulation of glycogen in LM of low RFI pigs was specifically located in the fast-twitch glycolytic IIBW fibers, which correspond to fibers containing IIb, IIb + IIx, or IIx myosin heavy chains. The difference in muscle glycogen content between RFI line pigs was more significant in the living animals (P = 0.0003) than at 30 min postmortem (P = 0.08). This was associated with a decreased ultimate pH (P = 0.001), and greater lightness of color (P = 0.002) and drip loss (P = 0.04) in LM of low than high RFI line pigs, suggesting that selection for reduced RFI may impair some meat quality traits, such as water-holding capacity. Pigs from the low RFI line exhibited a greater (P = 0.02) percentage of IIBW fibers in LM and tended (P < 0.10) to have less lipid β-oxidative capacity in LM, SM, and BFM. In contrast, no difference (P > 0.10) between lines was found for citrate synthase and lactate dehydrogenase activities, mitochondrial activity, and expression of genes coding for uncoupling proteins 2 and 3. Differences between RFI pigs in plasma leptin, cortisol, and thyroid hormone concentrations are presented and discussed. In conclusion, selection for low RFI influenced muscle properties in a way favoring muscle mass, but likely impairing meat quality.     

Adaptation of Equine Locomotor Muscle Fiber Types to Endurance and Intensive High Speed Training

Skeletal muscle constitutes the largest organ system in the mammalian body and is essential for movement and force generation. Muscle tissue has the unique ability to adapt and remodel with regular exercise. Adaptation of equine contractile apparatus to exercise training with a different character occurs at the structural to the cellular and molecular levels and depends on age, breed, and sex. In Andalusian and Arabian horses from 3 months to 24 years of age, it was found that the mean cross-sectional area occupied by IIA and IIX fibers was greater in stallions than in mares. In muscle of Dutch Warmblood foals from birth to 1 year of age, a significant number of fibers coexpress either developmental and type IIa or cardiac–alpha and type I myosin heavy chain (MyHC) isoforms. Endurance training results in increased mitochondrial density, capillary supply, changes in key metabolic enzymes, and increased maximal oxygen uptake and promotes a transition from type II to type I muscle fiber. In horses, prolonged aerobic exercise training has been shown to induce a further decline in the percentage of type IIx MyHC isoform expression and an increase of type I and IIa MyHC isoform expression. Short-duration, high-intensity exercise training stimulates type IIA and hybrid (IIA/IIX) fibers. Therefore, intensive high-speed trotting facilitates muscle fiber hypertrophy and increases the oxidative capacity of type IIX fibers.     

Age-Related Changes in Myosin Light and Heavy Chain Isoforms’ Composition in Gluteus Medius Muscle of Sedentary Akhal-Teke Horses

The primary purpose of the present study was to examine age dependence of gluteus medius muscle phenotypic plasticity with regard to myosin light chain (MyLC) and myosin heavy chain (MyHC) isoform expression of sedentary Akhal-Teke horses. A total of 23 Akhal-Teke horses (11 horses, 1.5 to 2.5-year-old; nine horses, 9 to 20-year-old; and three horses, 21 to 23.5-year-old) were studied. A muscle biopsy of depth 60 mm from the left gluteus medius muscle was obtained. MyHC and MyLC isoform pattern was determined in three age groups. The current study demonstrated that in the 21 to 23.5-year-old age group, the relative content of MyHC I isoform and MyLC 1_slow isoform decreased significantly. The results of the current study indicate that gluteus medius muscle of Akhal-Teke horses consists of a relatively high percentage of MyHC I and IIa isoforms, which together compose about 50% of MyHC. MyHC IIa and IIx isoforms together compose 88% to 94% of MyHC isoforms. Age-related decrease of the relative content of MyHC I and MyLC 1_slow isoforms in gluteus medius muscle starts in Akhal-Teke horses older than 20 years.     

Muscle fibre distribution in forelimb,hindlimb and trunk muscles in three bat species: The little Japanese horseshoe,greater horseshoe and Egyptian fruit

This study characterised muscle fibres in trunk, forelimb and hindlimb muscles of three bat species: little Japanese horseshoe (Rhinolophus cornutus), greater horseshoe (Rhinolophus ferrumequinum) and Egyptian fruit (Rousettus aegyptiacus). Twenty-seven muscles from trunk, forelimb and hindlimb were dissected, weighed and analysed by immunohistochemistry and sodium didecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and determined their cross-sectional areas (CSA). Results showed that Type IIa and Type IIa/x made the highest proportion of total muscle mass. Moderate proportion was formed by Type IIb. Type I and IIx appeared at very low levels in all bats. Type IIb was the only fibre type detected in patagial muscles in wing membrane of greater horseshoe while other fibre types were not observed. Type I muscle fibres were very few and appeared infrequently in fifteen muscles of Egyptian fruit and in only one muscle in each, greater horseshoe and little Japanese horseshoe. Type IIx was also detected in three muscles in greater horseshoe and only one muscle in Egyptian fruit but none in little Japanese horseshoe. The highest average CSA μm² was detected in Type IIb and values were 734.2μm² for LHB; 1537.9μm² for GHB and 1,720.9μm² for EFB. Lowest and undetermined values were observed for Type I and IIx. These data demonstrate that Type IIa, IIa/x and IIb form significant proportion of adult bat muscle mass and Type IIb is the largest fibre type. The distribution pattern is suggestive of specialised functions of the fibres in relation to orientation and speed of bats during flight.     

Influence of halothane genotype and body-weight on myosin heavy chain composition in pig muscle as related to meat quality

In an effort to understand the relationship between muscle fiber type, live weight, genotype, and PSE development, enzyme-linked immunosorbent analyses were used to evaluate myosin heavy chain (MyHC) isoform content in the longissimus muscle of pigs differing in halothane gene status (nn, homozygous mutant; Nn, heterozygous; NN, homozygous normal) that were slaughtered at three different weights (100, 120 and 140 kg). Pigs carrying the n gene (Nn and nn) exhibited more IIB MyHC and less slow type I MyHC than those pigs free of the n gene, while NN pigs had greater amounts of IIAX MyHC. The relative abundance of IIB and IIAX MyHC in muscle of all pigs studied was strongly negatively correlated (r=−0.834). Heavier pigs (140 kg) had the greatest amounts of slow and IIA MyHC. Across all genotypes, the relative abundance of IIB MyHC and muscle pH at 45 min postexsanguination (pH45) was negatively correlated (r=−0.418). In addition, the relative amount of slow was positively correlated with pH45 (r=0.386). Because muscle of homozygous nn positive pigs exhibited similar IIB/slow MyHC ratios to that of heterozygous Nn pigs, yet less desirable pH45 values and ultimate meat quality scores argues against a role of MyHC content per se in contributing to PSE development. However, these data do not preclude that those pigs with greater amounts of IIB MyHC are more ‘susceptible’ to adverse pork quality development than those pigs with less IIB MyHC.     

Differential response of cull cow muscles to the hypertrophic actions of ractopamine-hydrogen chloride

Ractopamine-HCl (RAC) is a beta-adrenergic agonist with variable effects on cattle performance and carcass variables. Cull cows fed RAC (200 mg . head(-1) . d(-1)) demonstrate an increased size of type I and II muscle fibers that does not translate into a larger ribeye area. The objective of this study was to examine the dose-dependent effects of RAC on cull cow muscle morphometrics. Eighty-eight cull beef cows representing 2 breed types (n = 44 each) were fed 0, 100, 200, and 300 mg . head(-1) . d(-1) of RAC for the last 28 d of a 54-d feeding period. On d 54, cows were slaughtered, and samples of the LM and semimembranosus muscle (SM) from 16 randomly selected carcasses (n = 4 per treatment) were taken for measurement of beta (2)-adrenergic receptors and type I, IIA, and IIX myosin heavy chain (MyHC) gene expression. Twenty-four hours postmortem, LM, SM, infraspinatus (INF), and vastus lateralis samples from 40 randomly selected carcasses (n = 10 per treatment) were obtained and frozen for immunohistochemical analysis. Muscle fiber cross-sectional area and diameter, MyHC isoform expression, and fiber-associated nuclei numbers were measured. Ractopamine dosage exhibited differential effects on muscle morphometrics and MyHC gene expression. Muscle fiber cross-sectional area and diameter were increased (P < 0.05) by RAC in INF type I and IIA fibers and SM type IIA fibers. Ractopamine increased (P < 0.05) MyHC type IIX mRNA and tended to increase (P < 0.10) beta(2)-adrenergic receptors in the SM; a change in mRNA abundance was not detected for either gene in the LM. Treatment with RAC decreased (P < 0.05) fiber-associated nuclei numbers in the INF, vastus lateralis, and LM but did not affect (P > 0.05) MyHC or beta-adrenergic receptor expression. These results indicate that cull cow feeding programs may consider supplementing RAC as a means of adding value to cuts within the chuck, such as the INF.     

The unique physiological features of the broiler pectoralis major muscle as suggested by the three-dimensional ultrastructural study of mitochondria in type IIb muscle fibers

Typical skeletal muscles are composed of mixed muscle fiber types, which are classified as slow-twitch (type I) and fast-twitch (type II) fibers, whereas pectoralis major muscles (PMs) in broiler chickens are 100% composed of type IIb fast-twitch fibers. Since metabolic properties differ among muscle fiber types, the combination of muscle fiber types is involved in physiological functions and pathological conditions in skeletal muscles. In this study, using serial block-face scanning electron microscopy, we compared three-dimensional (3D) mitochondrial properties in type IIb fibers in broiler PMs and those in type I fibers of broiler gastrocnemius muscles (GMs) heterogeneously composed of slow- and fast-twitch muscle fibers. In type I fibers in the GMs, elongated mitochondria with numerous interconnections to form a substantial network among myofibrils were observed. Along with lipid droplets sandwiched by mitochondria, these features are an adaptation to effective oxidative respiration and constant oxidative damage in slow-twitch muscle fibers. In contrast, type IIb fibers in the PMs showed small and ellipsoid-shaped mitochondria with few interconnections and no lipid droplets, forming a sparse network. The mitochondrial spatial network comprises of active mitochondrial dynamics to reduce mitochondrial damage; therefore, type IIb fibers possess physiologically low capacity to maintain mitochondrial wellness due to static mitochondrial dynamics. Based on 3D mitochondrial properties, we discussed the contrasting physiological functions between type I and IIb fibers and proposed a high contractile power and low stress resistance as unique physiological properties of broiler PMs.     

Within-litter variation in muscle fiber characteristics, pig performance, and meat quality traits

The objective of this study was to examine the intralitter variation in postnatal growth performance, meat quality, and muscle fiber characteristics when littermates were categorized by carcass weight. Thirty-nine litters were weaned at 4 wk of age and had free access to feed from 2 wk of age until slaughter. They were slaughtered by litter at an average BW of 104 +/- 14 kg, and six pigs per litter were selected for analysis: the heaviest- (HW), middle- (MW), and lightest-weight (LW) pig of each sex. Categorizing littermates in LW, MW, and HW pigs at the same age reflected the differences in postnatal growth rate within a litter; thus ADG, muscle mass, and muscle deposition rate differed across pig weight groups (P < 0.001). Also, the total DNA content was different among pig weight groups (P < 0.001) and reflected differences in muscle growth rate. The difference in muscle growth rate between LW and MW pigs could be explained by a larger (P < 0.05) mean fiber area (MFA) in MW pigs, whereas the number of muscle fibers was similar. Growth rate differences between MW and HW pigs could in part be explained by a higher number (P < 0.01) of equal-sized muscle fibers in HW pigs. The difference in MFA was due to a higher estimated DNA and RNA content per muscle fiber in MW and HW compared with LW pigs (P < 0.05). Pigment content was higher in MW and HW compared with LW pigs (P < 0.01), but no other measured meat quality traits were significantly different across pig weight groups. These results indicate that both the number and the growth rate of muscle fibers contribute to intralitter variation in postnatal growth performance.     

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.     

Low birth weight is associated with enlarged muscle fiber area and impaired meat tenderness of the longissimus muscle in pigs

The objective of this study was to determine the relationships between birth-weight-associated modifications in histological or chemical muscle characteristics and meat quality traits in pigs. At 68 d of age, Pietrain x (Large White x Landrace) female littermates were allocated into 2 groups on the basis of low birth weight (LW = 1.05 +/- 0.04 kg; n = 15) or high birth weight (HW = 1.89 +/- 0.02 kg; n = 15). Pigs were reared in individual pens with free access to a standard diet up to slaughter at approximately 112 kg of BW. During the growing-finishing period, LW and HW pigs had a similar daily feed consumption, whereas G:F was lower (P = 0.009) for LW pigs than for HW littermates. At final BW, LW pigs were 12 d older (P < 0.001) than HW littermates. Estimated lean meat content, relative proportions of loin and ham in the carcass, and weights of LM and semitendinosus muscle (SM) were decreased (P < 0.05) in LW pigs compared with HW pigs. Conversely, the LW pigs exhibited a fatter carcass, greater activity levels of fatty acid synthase and malic enzyme in backfat (n = 15 per group), and enlarged subcutaneous adipocytes (n = 8 per group) compared with the HW pigs. Similarly, lipid content was increased by 25% (P = 0.009), and mean adipocyte diameter was 12% greater (P = 0.008) in the SM from LW pigs compared with that from HW pigs, whereas lipid content did not vary in the LM of either group. Mean myofiber cross-sectional areas were 14% greater in the LM (P = 0.045) and the SM (P = 0.062) of LW pigs than of HW pigs. Conversely, the total number of myofibers was less (P = 0.003) in the SM of LW vs. HW pigs. There were no differences between groups for glycolytic potential at slaughter and rate and extent of postmortem pH decline in both muscles, as well as for LM drip losses. A trained sensory test panel judged the roast loin meat to be less tender (P = 0.002) in LW pigs relative to HW pigs. Scores for juiciness, flavor, flouriness, and fibrousness of meat did not differ between groups. Overall, negative but somewhat low correlation coefficients were found between LM tenderness score and ultimate pH (r = -0.36; P = 0.06) and between LM tenderness and mean cross-sectional area of myofibers (r = -0.34; P = 0.07). This study demonstrates a lower tenderness of meat from pigs that had a LW, partly as a result of their enlarged myofibers at market weight.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

肌球蛋白重链基因与肉品质的关系

肌纤维是构成肌肉的基本单位,肌纤维的特性直接影响猪肉品质.猪的肌纤维根据肌球蛋白重链(MyHC)的多态性可分为1、2a、2b和2x 4种类型,在代谢上分别与慢速氧化型、快速氧化型、快速酵解型和中间类型相对应.肌纤维的生成在分子水平上受到肌细胞生成素基因的精确调控,肌纤维的类型在生长过程中不断发生转化,并受遗传、生长、性别和营养等多因素的影响.     

The effects of selection of pigs on growth rate vs leanness on histochemical characteristics of different muscles

The microstructure of two type of muscles was studied in a selection experiment conducted with Dutch Large White pigs (boars and gilts) selected for either low backfat thickness (L-line) or fast growth (F-line). Second- and fourth-generation pigs were used to determine effects of selection on fiber type composition, fiber area, and capillary density in the longissimus lumborum (LL) and biceps femoris (BF) muscles. Immediately after slaughter samples were taken from the LL and BF muscles. The latter was divided into an inside (BFi) and outside (BFo) portion, which refer to the red and white portions of the biceps femoris. Serial sections were stained for ATPase (pH 4.60), succinate dehydrogenase (SDH), and alpha-amylase-periodic acid shiff (PAS) to determine fiber type and capillary density. The LL and BFo muscles had predominantly type IIBw fibers, whereas the BFi muscle had a 2 to 4 times higher amount of type I fibers. In most muscles there were more type I and fewer type IIBw fibers in F- than in L-line pigs (P < .05), except in the LL muscle of second-generation pigs and in the red part of the BF muscle of fourth-generation pigs. In both selection lines lower type I and higher type IIBw percentages were found in muscles from gilts than in those from boars (P < .05). Capillary density and fiber area of L- and F-lines showed minor differences, which could be explained by differences in weight and age of the pigs of both lines. The results suggest that selection for low backfat thickness in pig breeding compared with increased growth rate resulted in fewer oxidative and more glycolytic muscle fibers. The magnitude of the effect depended on muscle type and duration of the selection period.