Neuromuscular organization of avian flight muscle: Morphology and contractile properties of motor units in the pectoralis (pars thoracicus) of pigeon (Columba livia)

We used acid digestion and glycogen depletion to determine fascicle organization, fiber morphology, and physiological and anatomical features of individual motor units of an in-series muscle, the pectoralis (pars thoracicus) of the pigeon (Columba livia). Most fascicles are attached at one end to connective tissue. Average fiber length in the four regions examined range from 42% to 66% of average fascicle length. More than 65% of fibers are blunt at one end of a fascicle and taper intrafascicularly. Fibers with blunt–blunt endings range from 13% to 31% of the population in different regions; taper–taper fibers range from 2% to 17%. Pigeon pectoralis fibers are distinguished histochemically into fast-twitch glycolytic (FG) and fast-twitch oxidative-glycolytic (FOG) populations. Three units composed of FG fibers (FG units) contract more quickly than three units composed of FOG fibers (FOG units) (range 31–37 vs 47–62 msec), produce more tetanic force (0.11–0.32 vs 0.02–0.05 N) and are more fatigable (<18% initial force vs >50% after repeated stimulation). Most motor units are confined to one of the four muscle regions. Territory of two FOG units is <30% of parent fascicle length. Territories of other units spanned parent fascicles; most fibers in these units do not extend the full fascicle length. Compared to FG units, FOG units have lower maximum innervation ratios and density indices (ratio of depleted/total FOG fibers in territory 8–14% vs 58–76% for FG units). These differences support the hypothesis that FG units are organized to produce substantial force and power for takeoff, landing and other ballistic movements whereas FOG units are suited for sustained flight when power requirements are reduced. Implications of findings for understanding the control of in-series muscles and the use of connective tissue elastic elements during wing movements are discussed. J.Morphol. 236:179–208, 1998. © 1998 Wiley-Liss, Inc.     

Tail morphology in the Western Diamond‐backed rattlesnake,Crotalus atrox

The shaker muscles in the tails of rattlesnakes are used to shake the rattle at very high frequencies. These muscles are physiologically specialized for sustaining high‐frequency contractions. The tail skeleton is modified to support the enlarged shaker muscles, and the muscles have major anatomical modifications when compared with the trunk muscles and with the tail muscles of colubrid snakes. The shaker muscles have been known for many years to consist of three large groups of muscles on each side of the tail. However, the identities of these muscles and their serial homologies with the trunk muscles were not previously known. In this study, we used dissection and magnetic resonance imaging of the tail in the Western Diamond‐backed Rattlesnake, Crotalus atrox, to determine that the three largest muscles that shake the rattle are the M. longissimus dorsi, the M. iliocostalis, and the M. supracostalis lateralis. The architecture of these muscles differs from their serial homologs in the trunk. In addition, the rattlesnake tail also contains three small muscles. The M. semispinalis‐spinalis occurs in the tail, where it is a thin, nonvibratory, postural muscle that extends laterally along the neural spines. An additional muscle, which derives from fusion of the M. interarticularis inferior and M. levator costae, shares segmental insertions with the M. longissimus dorsi and M. iliocostalis. Several small, deep ventral muscles probably represent the Mm. costovertebrocostalis, intercostalis series, and transversohypapophyseus. The architectural rearrangements in the tail skeleton and shaker muscles, compared with the trunk muscles, probably relate to their roles in stabilizing the muscular part of the tail and to shaking the rattle at the tip of the tail. Based on comparisons with the tail muscles of a colubrid snake described in the literature, the derived tail muscle anatomy in rattlesnakes evolved either in the pitvipers or within the rattlesnakes. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.     

举重训练对男子肌比力、肌纤维的影响

安排9名有一定训练的青少年运动员参加10周举重训练。每周6次,每次8小时。训练前后用电子计算机—X线断层扫描技术(CT)测量大腿肌肉横断面积,股外肌针刺活检取样测定Ⅰ、Ⅱ型肌纤维％和面积。测定大腿伸膝肌最大随意等长力量。将结果(实验前、后)与对照组(13名无训练者)和优秀举重运动员(健将3人、一级4人)进行比较。结果表明,有训练人(实验前、后和优秀)与没有训练人(对照)在肌比力上存有差别。短、长期举重训练后,肌比力与Ⅱ型肌纤维％呈正相关。短、长期举重训练不曾改变肌纤维类型分布。举重训练对Ⅱ型肌纤维有选择性作用,且这个作用不受年龄的影响。结果还提示,肌比力的变化及肌比力与肌纤维之间的关系都会受运动项目专项特点的影响。     

Transgenic and tissue culture analyses of the muscle creatine kinase enhancer Trex control element in skeletal and cardiac muscle indicate differences in gene expression between muscle types

The muscle creatine kinase (MCK) gene is expressed at high levels only in differentiated skeletal and cardiac muscle. The activity of the cloned enhancer–promoter has previously been shown to be dependent on the Trex element which is specifically bound by a yet unidentified nuclear factor, TrexBF. We have further characterized the function of the Trex site by comparing wild-type and Trex-mutated MCK transgenes in five mouse skeletal muscles: quadriceps, extensor digitorum longus (EDL), soleus, diaphragm, and distal tongue, as well as in heart ventricular muscle. Several types of statistical analysis including analysis of variance (ANOVA) and rank sum tests were used to compare expression between muscle types and between constructs. Upon mutation of the Trex site, median transgene expression levels decreased 3- to 120-fold in the muscles examined, with statistically significant differences in all muscles except the EDL. Expression in the largely slow soleus muscle was more affected than in the EDL, and expression in the distal tongue and diaphragm muscles was affected more than in soleus. Median expression of the transgene in ventricle decreased about 18-fold upon Trex mutation. Transfections into neonatal rat myocardiocytes confirmed the importance of the Trex site for MCK enhancer activity in heart muscle, but the effect is larger in transgenic mice than in cultured cells.     

A mutation affecting larval muscle development in Drosophila melanogaster

The phenotypic analysis of a new spontaneous recessive lethal mutation of Drosophila melanogaster is described. The lethal(2)thin mutation maps at 85.6 on chromosome 2 and produces a characteristic long, thin puparium due to an inability to shorten the larval form prior to pupariation. Histological examination of larval muscles and behavioural studies support the hypothesis that the mutation affects the striated structure of the larval muscles in late larval stages. Lethality largely occurs due to an inability to perform the movements necessary for pupation, although there is evidence for larval and possibly embryonic lethal phases.     

Implication of basal lamina dependency in survival of Nrf2-null muscle stem cells via an antioxidative-independent mechanism

Nuclear factor erythroid 2–related factor 2 (Nrf2) is a master regulator for the induction of antioxidative genes and plays roles in diverse cellular functions. The roles of Nrf2 in muscle regeneration have been investigated, and both important and unimportant roles of Nrf2 for muscle regeneration have been reported. Here, using aged Nrf2-null and Nrf2–dystrophic double-null mice, we showed nonsignificant phenotypes in the muscle regeneration ability of Nrf2-null mice. In contrast with these results, strikingly, almost all Nrf2-null muscle stem cells (MuSCs) isolated by fluorescence-activated cell sorting died in vitro of apoptosis and were not rescued by antioxidative reagents. Although their proliferation was still impaired, the Nrf2-null MuSCs attached to myofibers activated and divided normally, at least in the first round. To elucidate these discrepancies of MuSCs behaviors, we focused on the basal lamina, because both in vivo and single myofiber culture allow MuSCs within the basal lamina to become activated. In a basal lamina–disrupted model, Nrf2-null mice exhibited remarkable regeneration defects without increased levels of reactive oxidative species in MuSCs, suggesting that the existence of the basal lamina affects the survival of Nrf2-null MuSCs. Taken together, these results suggest that the basal lamina compensates for the loss of Nrf2, independent of the antioxidative roles of Nrf2. In addition, experimental conditions might explain the discrepant results of Nrf2-null regenerative ability.     

Protein synthesis in skeletal muscle measured at different times during a 24 hour period

Six groups of 5 male rats (starting body weight 109 g) were allowed free access to a conventional rat diet. At 4 hourly intervals, starting at 10.00 h muscle protein synthesis was measured. By relating the weights of the gastrocnemius and soleus muscles to the initial body weights of the animals (i.e., at 09.30, day 1), a linear increase in muscle weight throughout the day was demonstrated. The fractional rate of muscle protein synthesis varied from 16.8% per day to 20.3% per day in gastrocnemius muscle and from 17.9% per day and 22.1% per day in the soleus. It was calculated that the maximum error incurred in estimating daily muscle protein synthesis by extrapolation of the value at any one time was 6% in gastrocnemius and 9% in soleus. It is concluded that calculations of the average rate of muscle protein degradation based on the difference between the rates of synthesis and deposition are generally valid in rats allowed free access to an adequate diet.     

Water compartments in living,glycerinated and fixed skeletal muscles of the frog

Summary The kinetics of water replacement with heavy water (deuterium oxide) in the gastrocnemius and sartorius muscles of the frog under isotonic conditions, studied both gravimetrically and by infrared photometry, reveals three water compartments: (i) non-exchangeable ( 80 ml/kg fresh weight), (ii) slowly exchanging ( 500 ml/kg fresh weight), (iii) rapid exchanging — extracellular ( 200 ml/kg fresh weight). Exposure to both glycerol and glutaraldehyde increases the permeability coefficients and the amount of rapid exchanging water; glutaraldehyde also increases the amount of nonexchangeable water. Approximately 90% of the water is kept in the tissue only by weak intermolecular forces, the energies of which amount to 1 kcal/mol. The amount of non-exchangeable water is equivalent to about six continuous adsorption layers covering the myofilaments. Approximately 70 % of the tissue water appears to be replaced by glutaraldehyde during standard fixation.     

Distinction between smooth muscle,fibroblasts and endothelial cells in culture by the use of fluoresceinated antibodies against smooth muscle actin

Summary FITC-labelled antibodies against native actin from chicken gizzard smooth muscle (Gröschel-Stewart et al., 1976) have been used to stain cultures of guinea-pig vas deferens and taenia coli, rabbit thoracic aorta, rat ventricle and chick skeletal muscle. The I-band of myofibrils of cardiac muscle cells and skeletal muscle myotubes stains intensely. In isolated smooth muscle cells, the staining is located exclusively on long, straight, non-interrupted fibrils which almost fill the cell. Smooth muscle cells which have undergone morphological dedifferentiation to resemble fibroblasts with both phase-contrast microscopy and electronmicroscopy still stain intensely with the actin antibody. In those muscle cultures which contain some fibroblasts or endothelial cells, the non-muscle cells are not stained with the actin antibody even when the reactions are carried out at 37° C for 1 h or after glycerination. Prefusion skeletal muscle myoblasts also do not stain with this antibody.It is concluded that the actin antibody described in this report is directed against a particular sequence of amino acids in muscle actin which is not homologous with non-muscle actin. The usefulness of this antibody in determining the origin of cells in certain pathological conditions such as atherosclerosis is discussed.This work was supported by the Life Insurance Medical Research Fund of Australia and New Zealand, the National Heart Foundation of Australia, the Deutsche Forschungsgemeinschaft and the Wellcome Trust (London). We thank Janet D. McConnell for excellent technical assistance     

Electron microscopy and 3D reconstruction of F-actin decorated with cardiac myosin-binding protein C (cMyBP-C)

Myosin-binding protein C (MyBP-C) is an ∼ 130-kDa rod-shaped protein of the thick (myosin containing) filaments of vertebrate striated muscle. It is composed of 10 or 11 globular 10-kDa domains from the immunoglobulin and fibronectin type III families and an additional MyBP-C-specific motif. The cardiac isoform cMyBP-C plays a key role in the phosphorylation-dependent enhancement of cardiac function that occurs upon β-adrenergic stimulation, and mutations in MyBP-C cause skeletal muscle and heart diseases. In addition to binding to myosin, MyBP-C can also bind to actin via its N-terminal end, potentially modulating contraction in a novel way via this thick-thin filament bridge. To understand the structural basis of actin binding, we have used negative stain electron microscopy and three-dimensional reconstruction to study the structure of F-actin decorated with bacterially expressed N-terminal cMyBP-C fragments. Clear decoration was obtained under a variety of salt conditions varying from 25 to 180 mM KCl concentration. Three-dimensional helical reconstructions, carried out at the 180-mM KCl level to minimize nonspecific binding, showed MyBP-C density over a broad portion of the periphery of subdomain 1 of actin and extending tangentially from its surface in the direction of actin's pointed end. Molecular fitting with an atomic structure of a MyBP-C Ig domain suggested that most of the N-terminal domains may be well ordered on actin. The location of binding was such that it could modulate tropomyosin position and would interfere with myosin head binding to actin.     

Myotube production in fast myotomal muscle is switched-off at shorter body lengths in male than female Atlantic halibut Hippoglossus hippoglossus (L.) resulting in a lower final fibre number

A sampling method is described to determine accurately the number of fast myotomal muscle fibres (N_F) in a large flatfish species, the Atlantic halibut Hippoglossus hippoglossus. An unusual feature of the fast myotomal muscle is the presence of internalized strips of slow muscle fibres. In fish of 1·5–3·5 kg (n = 24), the total cross‐sectional area (A_TC) of fast muscle was 18% greater in the dorsal than ventral myotomal compartments (P < 0·05), whereas there was no significant difference between left‐ and right‐hand sides of the body. Due the bilateral asymmetry, muscle blocks (5 × 5 × 5 mm) were prepared to systematically sample each myotomal quadrant (dorsal, ventral, left‐ and right‐side) and the diameters of 150 fast fibres measured per block. Smooth non‐parametric probability functions were fitted to a minimum of 800 measurements of fibre diameter per quadrant (n = 5). There were no significant differences in the distribution of muscle fibre diameters between myotomal compartments and therefore N_F could be estimated from a single quadrant. The number of blocks required to estimate N_F with a repeatability of ±2·5% increased from six at 300 g body mass to 17 at 96·5 kg, caused by variation within and between blocks. Gompertz curves were fitted to measurements of fibre number and fork length (L_F). The estimated final fibre number was 8·96 × 10⁵ (7·99–9·94 × 10⁵, 95% CI) for males and 1·73 × 10⁶ (1·56–1·90 × 10⁶, 95% CI) for female fish. The estimated L_F for cessation of fibre recruitment in the fast muscle of female fish (1775 mm) was almost twice that in males (810 mm), reflecting their greater ultimate body size.     

The relationship of creatine kinase variability with body composition and muscle damage markers following eccentric muscle contractions

[Purpose]

The purpose of the study was to investigate the relationship between CK variability and body composition and muscle damage markers following eccentric exercise.

[Methods]

Total 119 healthy male subjects were recruited to perform 50 eccentric contractions consisted of 2 sets of 25 contractions. Then, blood creatine kinase (CK) activity was analyzed to divide into three groups based on their CK activity levels. Maximum isometric strength (MIS), muscle soreness (SOR) and body composition data were obtained before and after exercise.

[Results]

The results showed that high CK responders had a significant decrease in MIS (p<0.001) and greater SOR (p<0.01) following eccentric exercise compared to low CK responders. Percent body fat was also higher in high responders compared to low responders (p=0.014). Peak CK activity was significantly correlated with MIS and SOR but no correlation with % body fat, muscle mass, and body mass index.

[Conclusion]

CK variability following eccentric exercise is closely related to MIS and SOR and % body fat may be a potent factor for CK variability.     

增加刺激频率不影响大鼠萎缩比目鱼肌强直收缩疲劳性

为观察模拟失重对大鼠比目鱼肌（soleus,SOL）与趾长伸肌（extensor digitorum longus,EDL）间断强直收缩功能的影响,以及对刺激频率的调节作用,采用离体骨骼肌条灌流技术,观测其产生强直收缩最大张力的最适刺激频率、疲劳性与疲劳后恢复过程。结果表明：对照组大鼠SOL强直收缩的最适刺激频率为60Hz,尾部悬吊1周大鼠SOL的最适刺激频率亦为60Hz,尾部悬吊2周后,其最适刺激频率增高为80Hz,4周后则为100Hz;在最适刺激频率作用下,悬吊大鼠SOL间断强直收缩的最大张力（Po）在悬吊1与2周未见改变,第4周才呈现显著性降低（P〈0．01）。间断强直收缩5min后,对照组大鼠SOL张力降低到22．8％Po：悬吊1、2与4周组疲劳性均增加,与其同步对照组相比均有显著性差异（P〈0．01）。疲劳性强直收缩后,在20min内对照大鼠SOL张力基本恢复到疲劳前水平,而悬吊1、2与4周组则不能完全恢复（P〈0．05）。对照组大鼠EDL的最适刺激频率为120Hz,悬吊1、2与4周组EDL的最适刺激频率、疲劳性以及疲劳后恢复过程均未发生改变。以上结果提示,增加刺激频率可对悬吊1与2周大鼠SOL强直收缩最大张力的降低有代偿作用,但不能代偿悬吊4周大鼠SOL最大收缩张力的降低,亦不影响悬吊大鼠SOL间断强直收缩疲劳性的增加与疲劳后恢复的减缓。     

Ciliary muscle contraction force and trapezius muscle activity during manual tracking of a moving visual target

Previous studies have shown an association of visual demands during near work and increased activity of the trapezius muscle. Those studies were conducted under stationary postural conditions with fixed gaze and artificial visual load. The present study investigated the relationship between ciliary muscle contraction force and trapezius muscle activity across individuals during performance of a natural dynamic motor task under free gaze conditions. Participants (N = 11) tracked a moving visual target with a digital pen on a computer screen. Tracking performance, eye refraction and trapezius muscle activity were continuously measured. Ciliary muscle contraction force was computed from eye accommodative response. There was a significant Pearson correlation between ciliary muscle contraction force and trapezius muscle activity on the tracking side (0.78, p < 0.01) and passive side (0.64, p < 0.05). The study supports the hypothesis that high visual demands, leading to an increased ciliary muscle contraction during continuous eye–hand coordination, may increase trapezius muscle tension and thus contribute to the development of musculoskeletal complaints in the neck–shoulder area. Further experimental studies are required to clarify whether the relationship is valid within each individual or may represent a general personal trait, when individuals with higher eye accommodative response tend to have higher trapezius muscle activity.     

Surface electromyography activity of the rectus abdominis,internal oblique,and external oblique muscles during forced expiration in healthy adults

We aimed to characterize rectus abdominis, internal oblique, and external oblique muscle activity in healthy adults under expiratory resistance using surface electromyography. We randomly assigned 42 healthy adult subjects to 3 groups: 30%, 20%, and 10% maximal expiratory intraoral pressure (PEmax). After measuring 100% PEmax and muscle activity during 100% PEmax, the activity and maximum voluntary contraction of each muscle during the assigned experimental condition were measured. At 100% PEmax, the external oblique (p < 0.01) and internal oblique (p < 0.01) showed significantly elevated activity compared with the rectus abdominis muscle. Furthermore, at 20% and 30% PEmax, the external oblique (p < 0.05 and < 0.01, respectively) and the internal oblique (p < 0.05 and < 0.01, respectively) showed significantly elevated activity compared with the rectus abdominis muscle. At 10% PEmax, no significant differences were observed in muscle activity.Although we observed no significant difference between 10% and 20% PEmax, activity during 30% PEmax was significantly greater than during 20% PEmax (external oblique: p < 0.05; internal oblique: p < 0.01). The abdominal oblique muscles are the most active during forced expiration. Moreover, 30% PEmax is the minimum intensity required to achieve significant, albeit very slight, muscle activity during expiratory resistance.     

Ultrastructure of a molluscan smooth muscle during phasic contractions and in the relaxed state

Tension and X-ray diffraction patterns are not always correlated in the smooth anterior retractor muscle (ABRM) of Mytilus edulis. The muscle produces equatorial intensity profiles of X-ray diffraction patterns corresponding to either a relaxed or a contracted structure. During phasic contractions, comprising a contracted as well a a relaxed phase, the diffracted intensity on the equator at 0.003 A^?1 changes within the first 10s after onset of stimulation. The tension reaches a maximum after about the same time. The time dependence of this intensity change during phasic contraction has been measured. It shows that the tension decays within 10s, but the relaxed structure needs 30–40 s to reestablish. There is no difference between the observed intensities from the tonic and phasic contracted states. Inactivated muscles with minimum tension, normally termed relaxed, can have either a “contracted” or a relaxed structure.     

Krüppel样因子在肌肉组织中的功能研究进展

Krüppel样因子(Krüppel-like factors, KLFs)是一类C-末端含有3个C₂H₂锌指结构的转录因子,N-末端为转录调控结构域,能够结合多种特异蛋白质,介导转录调控。目前在人体基因组中共发现18种KLFs,它们在多种类型人类细胞的分化、表型维持和生理功能调控中发挥重要作用。多个KLFs参与了对人和动物的心肌、平滑肌和骨骼肌的发育和功能的调控。在心肌中,KLF4、KLF10、KLF11和KLF15参与心肌肥大的负调控,KLF6参与调控心脏纤维化,KLF13调控胚胎时期的心肌发育。在血管平滑肌中,KLF4受促增殖或促分化因子调控,介导调控血管平滑肌表型转换;KLF5促进血管平滑肌增殖,KLF8和KLF15抑制血管平滑肌增殖。在骨骼肌中,KLF2、KLF3、KLF4、KLF10和KLF15调控骨骼肌发育,此外,KLF15是肌肉组织能量代谢的调节因子。本文综述了KLFs在心肌、平滑肌和骨骼肌中的功能研究进展,为进一步揭示KLFs在肌肉组织中的作用和肌肉相关疾病的分子机制提供参考。     

Active muscle migration during insect metamorphosis

The R1 abdominal retractor muscles of the insect Tenebrio molitor change position during the course of metamorphosis. These muscles detach from the epidermal tendon cells at their anterior ends, and migrate in a posterior direction, parallel to the body axis, to form completely new attachments shortly before adult emergence. Movement is preceded by the loss of sarcomere structure, and the muscles migrate in a partially dedifferentiated condition, closely accompanied by satellite cells and haemocytes. Movement appears to result from the extension of muscle processes towards the epidermis posterior to the larval attachment sites, which contact reciprocal processes extended from the epidermis. Contacts at the new posterior sites are then reinforced, and relinquished at the anterior. This cycle is subsequently repeated. It is envisaged that migration ceases when the muscles encounter a contour in the epidermal gradient known to specify the position of the adult muscle attachment sites. This positional information may be encoded in the epidermal basal lamina. The muscles then redifferentiate, with concurrent differentiation of new epidermal tendon cells. Development of adult muscle attachments appears to require reciprocal morphogenetic interactions between muscle and epidermis.     

The fine structure of red and white myotomal muscle fibres of the coalfish (Gadus virens)

Summary The fine structure of the red and white myotomal muscles of a marine teleost, the coalfish Gadus virens, has been examined and ultrastructural measurements and analyses carried out. The sarcomere lengths of the red and white fibres were found to be 1.60 minimum, 1.82 maximum and 1.70 minimum, 1.85 maximum, respectively. No significant difference was found between the red and white fibres in their percentage of sarcoplasmic reticulum and T system. Both were found to have regularly occurring triads at the Z disk level, to have distinctive M lines and to be multiply innervated. Ultrastructurally the two fibres can be distinguished by the thicker Z line and more abundant mitochondria of the red fibre, and by the ribbon-shaped peripheral myofibrils of the white fibres. The structure of the fibres in these two types of muscle is discussed in relation to their possible role in swimming.This work was supported by a research grant from the National Environmental Research Council.