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.     

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.     

The fiber composition of the semitendinosus muscle of the rabbit

Summary The semitendinosus muscle of the rabbit is composed of a homogeneous fiber population. Fiber typing was carried out by means of light and electron microscopy according to which this muscle exhibits structural features that are either characteristic for both red and white fibers, or that do not allow for any classification according to the A-, B-, C-fiber system. Hence the long-held assumption that the semitendinosus muscle of the rabbit represents a classical paradigm of a red muscle should be revised in light of the present results.     

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.     

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.     

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.     

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.     

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     

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.     

A comparative analysis of the vasomotor responses and innervation of small arteries in rat locomotor and respiratory muscles

Characteristics of the small arteries (with a diameter of 200-250 μm) feeding the medial gastrocnemius muscle and diaphragm were studied. Recording of the mechanical activity of ring segments under isometric conditions demonstrated that, similar to other arteries feeding the muscles with a high content of slow fibers, the diaphragm arteries are highly sensitive to adrenoceptor agonists and acetylcholine. The differences in the endothelium-dependent relaxation in response to acetylcholine were retained in the presence of L-NAME and diclofenac. The diaphragm and gastrocnemius arteries similarly responded to serotonin. On the other hand, a high innervation density was characteristic of the diaphragm arteries unlike the arteries of other slow muscles. The density of adrenergic nerve plexus in the diaphragm arteries was considerably higher than in the gastrocnemius arteries. The results suggest that the characteristics of small diaphragm arteries are determined not only by the oxidative capacity of diaphragm muscle fibers, but also by the fact that this is a respiratory muscle.     

Role of innervation on the embryonic development of skeletal muscle

Summary The extent to which the motor innervation regulates the embryonic development of skeletal muscle was investigated by comparing changes in normal, aneural, and paralyzed superior oblique muscle of the duck embryo. The muscle was made aneural by permanently destroying the trochlear motor neurons with electrocautery on day 7 i.e., three days prior to innervation. Embryos were paralyzed by daily application of -bungarotoxin onto the chorioallantoic membrane from day 10 onwards. The differentiation of myoblasts and myotubes in the aneural muscle was severely affected and did not progress to the myofiber stage. A mass of dead cells in the aneural muscle was replaced by connective tissue. Although the differentiation of myoblasts and myotubes was also retarded in the paralyzed muscle, numerous muscle cells progressed to the myofiber stage. Neuromuscular junctions of normal ultrastructure were seen in all paralyzed muscles. Degeneration of some cells in the paralyzed muscle occurred but there was no evidence of a massive wave of cell death similar to that observed in the aneural muscle. These observations suggest that both the trophic factors from the nerve and the nerve-evoked muscle activity are essential for the execution of the developmental program of the muscle. Trophic factors may play a larger role in differentiation, and maintenance of the muscle than muscle activity.Supported by a grant from the Muscular dystrophy Association and a grant from NIHWe are grateful to Beth McBride and Greg Oblak for their technical assistance     

Terminal sprouting in the sternocostalis muscle of the rat

Summary Partial denervation of the sternocostalis muscle was achieved by sectioning two out of five of its intercostal nerves. The terminal sprouting response was markedly increased compared to that found following section of only one nerve. The increase in the response was greater for B type than for C type end plates, although B type end plates appear unable to produce terminal sprouts longer than 20 m after partial denervation.Double nerve section allowed terminal sprouts from C type end plates to increase in length up to three days, after which time they appeared to retract. It is postulated that the onset of collateral sprouting resulted in reinnervation of empty end plate sites and hence removed the target for terminal sprouts.     

Ultrastructure of the muscles of the dorsal diaphragm in Locusta migratoria

Summary The alary muscles of Locusta migratoria adults make up the major tissue of the dorsal diaphragm which separates pericardial and perivisceral sinuses in the abdomen. The alary muscles are striated with a sarcomere at rest measuring about 9 m. The Z-line has a staggered-beaded arrangement with A-bands and I-bands readily discernable. Thick myofilaments are surrounded by 10 or more thin filaments. The sarcoplasm has few mitochondria near the area of the Z-line, dyads are present and sarcoplasmic reticulum is poorly developed. Axons which innervate the alary muscle are either contained within invaginated folds of the sarcolemma of the muscle cells or the muscle cells send finger-like projections to envelop the axons. The synaptic terminals contain synaptic vesicles between 40 and 45 nm in diameter and a few electron-dense granules near or less than 170 nm in diameter. Away from synaptic terminals the axon profiles show few or no granules. The axons are accompanied everywhere by well-developed glial cells. This then is not typical neurosecretomotor innervation, however, the presence of electron-dense granules suggests the possibility of peptidergic neurotransmission.     

Haploinsufficiency of myostatin protects against aging‐related declines in muscle function and enhances the longevity of mice

The molecular mechanisms behind aging-related declines in muscle function are not well understood, but the growth factor myostatin (MSTN) appears to play an important role in this process. Additionally, epidemiological studies have identified a positive correlation between skeletal muscle mass and longevity. Given the role of myostatin in regulating muscle size, and the correlation between muscle mass and longevity, we tested the hypotheses that the deficiency of myostatin would protect oldest-old mice (28–30 months old) from an aging-related loss in muscle size and contractility, and would extend the maximum lifespan of mice. We found that MSTN^+/− and MSTN^−/− mice were protected from aging-related declines in muscle mass and contractility. While no differences were detected between MSTN^+/+ and MSTN^−/− mice, MSTN^+/− mice had an approximately 15% increase in maximal lifespan. These results suggest that targeting myostatin may protect against aging-related changes in skeletal muscle and contribute to enhanced longevity.     

Immunohistochemical Analysis of the Effects of Cross-innervation of Murine Thyroarytenoid and Sternohyoid Muscles

This work uses cross-innervation of respiratory muscles of different developmental origins to probe myogenic and neurogenic mechanisms regulating their fiber types. The thyroarytenoid (TA) originates from the sixth branchial arch, whereas the sternohyoid (SH) is derived from somitic mesoderm. Immunohistochemical analysis using highly specific monoclonal antibodies to myosin heavy chain (MyHC) isoforms reveals that normal rat SH comprises slow, 2a, 2x, and 2b fibers, as in limb fast muscles, whereas the external division of the TA has only 2b/eo fibers coexpressing 2B and extraocular (EO) MyHCs. Twelve weeks after cross-innervation with the recurrent laryngeal nerve, the SH retained slow and 2a fibers, greatly increased the proportion of 2x fibers, and their 2b fibers failed to express EO MyHC. In the cross-innervated TA, the SH nerve failed to induce slow and 2A MyHC expression and failed to suppress EO MyHC expression in 2b/eo fibers. However, 2x fibers amounting to 4.2% appeared de novo in the external division of the TA. We conclude that although MyHC gene expression in these muscles can be modulated by neural activity, the patterns of response to altered innervation are largely myogenically determined, thus supporting the idea that SH and TA differ in muscle allotype. (J Histochem Cytochem 58:1057–1065, 2010)     

Quantitative effects of thermal injury and insulin on the metabolism of the skeletal muscle using the perfused rat hindquarter preparation

Injury from a severe burn or trauma can propel the body into a hypermetabolic state that can lead to the significant erosion of lean muscle mass. Investigations describing this process have been somewhat limited due to the lack of adequate experimental models. Here we report the use of a perfused rat hindquarter preparation to study the consequences of a moderate burn injury (approximately 20% total body surface area), with or without the addition of exogenous insulin (12.5 mU/mL), on the fluxes of major metabolites across the isolated skeletal muscle. The metabolic flux data was further analyzed using metabolic flux analysis (MFA), which allows for the estimation of the impact of these conditions on the intracellular muscle metabolism. Results indicate that this model is able to capture the increased rate of proteolysis, glutamine formation, and the negative nitrogen balance associated with the burn-induced hypermetabolic state. The inclusion of exogenous insulin resulted in significant changes in several fluxes, including an increase in the metabolism of glucose and the flux through the pentose phosphate pathway, as well as a reduction in the metabolism of glutamine, alanine, and leucine. However, insulin administration did not affect the nitrogen balance or the rate of proteolysis in the muscle, as has been suggested using other techniques. The use of the perfused hindquarter model coupled with MFA is a physiologically relevant and experimentally flexible platform for the exploration of skeletal muscle metabolism under catabolic conditions, and it will be useful in quantifying the specific metabolic consequences of other therapeutic advances.     

The morphology and cell culture of the striated musculature of the rat azygos vein

Summary The azygos vein of the rat can be divided into three regions: 1) The proximal cardiac region, where the wall is composed of two and sometimes three layers of cardiac muscle and a thin discontinuous layer of smooth muscle cells. Vascular casts of this region demonstrate layers of capillaries closely following the orientation of the cardiac fibres. 2) A transitional zone, where both cardiac and smooth muscle cells interdigitate. In this zone, close associations between smooth muscle and cardiac muscle cells can be observed, however, gap junctions do not appear to be present. 3) Beyond this transitional zone the vessel resembles a typical thin-walled vein.The cells of the media of the entire length of azygous vein have been isolated and grown in culture and two separate viable populations identified corresponding to smooth and cardiac muscle.     

肥胖诱导的骨骼肌萎缩机制研究进展

骨骼肌是人体氨基酸和蛋白质的主要贮存、代谢库,其正常功能和代谢过程受到多种病理因素的影响。骨骼肌萎缩发生于骨骼肌稳态严重失衡状态下,对患者生活和社会医疗造成了沉重负担。近年来,由于世界肥胖人群数量激增,肥胖诱导的骨骼肌萎缩正日益成为公共卫生的严峻挑战之一。肥胖诱导的骨骼肌萎缩过程涉及多种信号分子或通路的改变,如泛素蛋白酶系统、自噬溶酶体系统、胰岛素/IGF1-PI3K-Akt、肌肉生长抑制素、白细胞介素-6、肿瘤坏死因子等;这些信号分子或通路在肥胖状态下被激活或抑制后,可共同影响蛋白质合成/分解平衡进而造成骨骼肌萎缩。基于上述信号分子或通路,系统总结并讨论了肥胖诱导的骨骼肌萎缩机制,以期为寻找缓解/治疗肥胖诱导的肌萎缩靶点和进一步开发利用天然植物化学物提供理论依据。     

Effect of thyroid hormones on the gene expression of calcium transport systems in rat muscles

It has been previously shown that modification of thyroid hormone levels have a profound impact on cardiac function, predominantly through a direct regulation of the sarcoplasmic reticulum protein levels. Nevertheless, little is known about the regulation of calcium transport systems in skeletal muscle due to the altered concentration of thyroid hormones. Thus, the goal of our study was to find out whether altered thyroid status could change the gene expression of the Na(+)/Ca(2+) exchanger (NCX), the inositol 1,4,5-trisphosphate (IP(3)) receptors and ryanodine receptors (RyRs) in slow and fast skeletal muscles of rats. A hyperthyroid state was maintained in rats by triiodothyronine (T(3)) administration, while methimazole was employed for inducing hypothyroidism. After a period of 2-10 months of T(3) treatment we observed a significant increase in mRNA levels of the NCX, RyRs and IP(3) receptors. This increase was more pronounced in the slow soleus than in the fast extensor digitorum longus (EDL) muscle. It is tempting to speculate that thyroid hormones also alter calcium concentration and thus influence the process of excitation-contraction coupling in the skeletal muscle.