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Measurement of muscle hardness using a hardness meter: application to the masseter and temporal muscles and reproducibility of measurement.

This study was designed to evaluate the intra-examiner and inter-examiner reliability of measurements of masticatory muscle hardness and to confirm that the muscle hardness increases with contraction using a commercially available muscle meter. Twenty healthy asymptomatic female subjects participated in this study. Hardness was expressed as numerical relative values (0-100). First, muscle hardness was measured at a standardized point located in the masseter muscle and temporal muscle in a randomized order by two examiners, and again by one of the same examiners after ten minutes for the reproducibility study. Then the muscle hardness was measured at each point for 0 kgf, 3 kgf, 6 kgf, and 9 kgf levels of bite force. As a result, intraclass correlation coefficient (ICC) analysis revealed good intra-examiner reliability in the masseter muscle (ICC = 0.711), good intra-examiner reliability in the temporal muscle (ICC = 0.643), good inter-examiner reliability in the masseter muscle (ICC = 0.631), and unacceptable inter-examiner reliability in the temporal muscle (ICC = 0.008). Also, our results showed that muscle hardness increased with contraction, and relationships with a slope of 1.229, a y-intercept of 62.513, and a correlation coefficient of 0.448 were observed in the masseter muscle. However, no correlation was found between muscle hardness and bite force in the temporal muscle. The findings indicate that measurement of hardness provides reliable physiological information about the masseter muscle in this setting.

Adolescent↗

Comprehensive expression profiling by muscle tissue class and identification of the molecular niche of extraocular muscle.

Muscle tissue is an elegant model for biologic integration of structure with function and is frequently affected by a variety of inherited diseases. Traditional muscle classes--skeletal, cardiac, and smooth--share basic aspects of contractile and energetics mechanisms but also have distinctive role-specific adaptations. We used large-scale oligonucleotide microarrays to broaden knowledge of the adaptive expression patterns underlying muscle tissue differences and to identify transcript subsets that are most likely to represent candidate disease genes. Using stringent analysis criteria, we found >or=95 transcripts, which were preferentially expressed by each muscle class and were validated by inclusion of known muscle class-specific and inherited disease-related genes. Differentially expressed transcripts not previously identified as class-specific extend understanding of muscle class transcriptomes and may represent novel muscle-specific disease genes. We also analyzed the expression profile of extraocular muscle, which is divergent from other skeletal muscles, in the broader context of all major muscle classes. Data show that the extraocular muscle phenotype results from the combination of tissue-specific transcripts, novel expression levels of skeletal muscle transcripts, and partial sharing of gene expression patterns with cardiac and smooth muscle. These, and additional proteomic data, establish that extraocular muscle does not constitute a distinctive muscle class but that it does occupy a novel niche within the skeletal muscle class.

Animals↗

Beta 2-agonist administration reverses muscle wasting and improves muscle function in aged rats.

The beta2-adrenoceptor agonist (beta2-agonist) fenoterol has potent anabolic effects on rat skeletal muscle. We conducted an extensive dose-response study to determine the most efficacious dose of fenoterol for increasing skeletal muscle mass in adult rats and used this dose in testing the hypothesis that fenoterol may have therapeutic potential for ameliorating age-related muscle wasting and weakness. We used adult (16-month-old) rats that had completed their growth and development, and old (28-month-old) rats that exhibited characteristic muscle wasting and weakness, and treated them daily with either fenoterol (1.4 mg kg(-1), i.p), or saline vehicle, for 4 weeks. Following treatment, functional characteristics of fast-twitch extensor digitorum longus (EDL) and predominantly slow-twitch soleus muscles of the hindlimb were assessed in vitro. Untreated old rats exhibited a loss of skeletal muscle mass and a decrease in force-producing capacity, in both fast and slow muscles, compared with adult rats (P < 0.05). However, there was no age-associated decrease in skeletal muscle beta-adrenoceptor density, nor was the muscle response to chronic beta-agonist stimulation reduced with age. Thus, muscle mass and force-producing capacity of EDL and soleus muscles from old rats treated with fenoterol was equivalent to, or greater than, untreated adult rats. The increase in mass and strength was attributed to a non-selective increase in the cross-sectional area of all muscle fibre types, in both the EDL and soleus. Fenoterol treatment caused a small increase in fatiguability due to a decrease in oxidative metabolism in both EDL and soleus muscles, with some cardiac hypertrophy. Further studies are needed to fully separate the desirable effects on skeletal muscle and the undesirable effects on the heart. Nevertheless, our results demonstrate that fenoterol is a powerful anabolic agent that can restore muscle mass and strength in old rats, and provide preliminary evidence of therapeutic potential for age-related muscle wasting and weakness.

Adrenergic beta-2 Receptor Agonists↗

Effect of altering starting length and activation timing of muscle on fiber strain and muscle damage.

Muscle strain injuries are some of the most frequent injuries in sports and command a great deal of attention in an effort to understand their etiology. These injuries may be the culmination of a series of subcellular events accumulated through repetitive lengthening (eccentric) contractions during exercise, and they may be influenced by a variety of variables including fiber strain magnitude, peak joint torque, and starting muscle length. To assess the influence of these variables on muscle injury magnitude in vivo, we measured fiber dynamics and joint torque production during repeated stretch-shortening cycles in the rabbit tibialis anterior muscle, at short and long muscle lengths, while varying the timing of activation before muscle stretch. We found that a muscle subjected to repeated stretch-shortening cycles of constant muscle-tendon unit excursion exhibits significantly different joint torque and fiber strains when the timing of activation or starting muscle length is changed. In particular, measures of fiber strain and muscle injury were significantly increased by altering activation timing and increasing the starting length of the muscle. However, we observed differential effects on peak joint torque during the cyclic stretch-shortening exercise, as increasing the starting length of the muscle did not increase torque production. We conclude that altering activation timing and muscle length before stretch may influence muscle injury by significantly increasing fiber strain magnitude and that fiber dynamics is a more important variable than muscle-tendon unit dynamics and torque production in influencing the magnitude of muscle injury.

Animals↗

Stiffness of cat soleus muscle and tendon during activation of part of muscle.

Experiments have been carried out on the soleus muscle and its tendon in the anesthetized cat. Measurements of isometric tension and muscle stiffness were made during contraction of whole or part of the muscle in response to stimulation of ventral root filaments. In an attempt to determine the distribution of tension in different portions of the tendon during activation of only part of the muscle, the free tendon of insertion was split longitudinally into two halves and a strain gauge attached to each piece. From a large number of measurements, it was found that the mean fraction of tension recorded in one-half of the tendon remained about the same, over a wide range of tensions. However, the scatter of values, which increased as the portion of muscle contracting was reduced, was greater than expected if muscle fibers were randomly distributed throughout the muscle. Measurements of muscle and tendon stiffness were made from length and tension changes during stretch of the actively contracting muscle. Ventral root stimulation that engaged 20% or more of the muscle yielded a value for tendon compliance (0.09 mm/N), which was the same as for stimulating the whole muscle. This result suggested that for contraction of portions as small as 20% of the muscle, fibers were effectively attached to the whole tendon, indicating that tendinous attachments of individual muscle fibers ran independent of one another over only a short distance and were bound together over most of their remaining course. It was concluded that groups of muscle fibers selected by stimulation of ventral root filaments are not entirely randomly distributed throughout the muscle. However, for groups representing larger fractions of the total tension, (greater than 20%) the distribution is uniform enough and the connections between their tendinous attachments firm enough for the force applied by such a group to act through a tendon compliance, which is the same as that seen by the whole muscle.

Achilles Tendon↗

Self-reinnervated cat medial gastrocnemius muscles. II. analysis of the mechanisms and significance of fiber type grouping in reinnervated muscles.

1. The technique of glycogen depletion was used to determine whether regenerating motor axons reestablish the normal regionalization of motor units (MUs) in the cat medial gastrocnemius (MG) muscle, 2) whether the extent of clumping between MU fibers and/or type grouping of muscle fibers progressively increases with a decrease in reinnervated MU numbers, and 3) whether the pattern of innervation can explain why MUs fail to increase significantly in size when the cut nerve is sutured directly to the muscle, even when few axons make functional connections. 2. Distributions of MU fibers were analyzed in 5 normal and 14 reinnervated cat MG muscles 4.5-16 mo after sectioning of its nerve and suturing of the proximal end to the distal nerve sheaths (N-N suture) or directly to the muscle fascia (N-M suture). Muscle unit distributions were quantified according to location, territory size, density, and extent of clumping between fibers from the same MU. 3. Normal MU fibers were regionalized within five regions along the muscle's longitudinal and transverse axes. Reinnervated MUs were located within similar regions, indicating that regenerating axons follow the major proximal nerve branches to restore normal compartmentalization. 4. Muscle unit fibers were diffusely scattered within discrete MU territories in normal muscles. Territory size tended to increase with MU size, whereas density of muscle unit fibers within the territory decreased. 5. Territories increased with MU size after N-N suture but were smaller and showed little size variation after N-M suture. The extent of muscle unit fiber clumping was inversely related to the number of reinnervated MUs. On average, the extent of clumping was substantially higher in muscles reinnervated after N-M suture. These results indicate that distal nerve sheaths facilitate proximal axon branching, which establishes MU territory size. Once the territory is established, motor axons branch distally to increase MU size, which in turn compensates for reduced MU numbers. 6. Muscles reinnervated by < 80% of the MUs exhibited fiber type grouping of type I fibers, and on average the extent of clumping was substantially higher in muscles reinnervated after N-M suture. With less innervation, type grouping increased inversely with the number of reinnervated MUs. However, for a similar number of MUs, type I fiber type grouping was substantially higher in muscle reinnervated after N-M suture. Type grouping therefore reflects muscle unit fiber clumping under conditions where MU size increased (N-N suture) or MU territory size decreased (N-M suture).

Animals↗

Vascular smooth muscle cells spontaneously adopt a skeletal muscle phenotype: a unique Myf5(-)/MyoD(+) myogenic program.

Smooth and skeletal muscle tissues are composed of distinct cell types that express related but distinct isoforms of the structural genes used for contraction. These two muscle cell types are also believed to have distinct embryological origins. Nevertheless, the phenomenon of a phenotypic switch from smooth to skeletal muscle has been demonstrated in several in vivo studies. This switch has been minimally analyzed at the cellular level, and the mechanism driving it is unknown. We used immunofluorescence and RT-PCR to demonstrate the expression of the skeletal muscle-specific regulatory genes MyoD and myogenin, and of several skeletal muscle-specific structural genes in cultures of the established rat smooth muscle cell lines PAC1, A10, and A7r5. The skeletal muscle regulatory gene Myf5 was not detected in these three cell lines. We further isolated clonal sublines from PAC1 cultures that homogeneously express smooth muscle characteristics at low density and undergo a coordinated increase in skeletal muscle-specific gene expression at high density. In some of these PAC1 sublines, this process culminates in the high-frequency formation of myotubes. As in the PAC1 parental line, Myf5 was not expressed in the PAC1 sublines. We show that the PAC1 sublines that undergo a more robust transition into the skeletal muscle phenotype also express significantly higher levels of the insulin-like growth factor (IGF1 and IGF2) genes and of FGF receptor 4 (FGFR4) gene. Our results suggest that MyoD expression in itself is not a sufficient condition to promote a coordinated program of skeletal myogenesis in the smooth muscle cells. Insulin administered at a high concentration to PAC1 cell populations with a poor capacity to undergo skeletal muscle differentiation enhances the number of cells displaying the skeletal muscle differentiated phenotype. The findings raise the possibility that the IGF signaling system is involved in the phenotypic switch from smooth to skeletal muscle. The gene expression program described here can now be used to investigate the mechanisms that may underlie the propensity of certain smooth muscle cells to adopt a skeletal muscle identity.(J Histochem Cytochem 48:1173-1193, 2000)

Animals↗

Genome-Wide Association Study on Muscle Stiffness Identified Novel Locus for Predisposition to Muscle Strain Injury.

PURPOSE: We aimed to screen the entire genome for genetic variants associated with passive muscle stiffness, which has been suggested as a risk factor for muscle strain injury. METHODS: This genome-wide association study (GWAS) on passive muscle stiffness included 350 physically active young Japanese individuals. Three hamstring constituents were measured using ultrasound shear wave elastography. Skeletal muscle transcriptomes were compared across the genotypes of GWAS-identified variants in 48 healthy Japanese individuals. Association between GWAS-identified variants and history of muscle strain injury was examined in 1428 Japanese athletes. RESULTS: Two loci on chromosome 11 demonstrated a genome-wide significant association with passive muscle stiffness of the biceps femoris long head (rs12807854 T/C: P = 5.19 &#xd7; 10 -10 , rs78405694 T/C: P = 2.09 &#xd7; 10 -8 ; linear regression analysis adjusted for sex, age, and stretching exercise habits). Skeletal muscle RNA sequencing revealed significantly elevated expression of extracellular matrix-related genes in muscles carrying stiffness-increasing alleles of these variants. Among athletes, rs12807854 T/C was significantly associated with a history of muscle strain injury ( P = 0.0254; logistic regression analysis adjusted for age, sex, competitive level, and main sport). Carriers of the C allele, associated with increased muscle stiffness, exhibited a heightened risk of muscle strain injury (odds ratio = 1.62; 95% confidence interval = 1.06-2.47 per C allele increase). By contrast, rs78405694 did not show a significant association with muscle strain injury in this population. CONCLUSIONS: A novel locus associated with passive muscle stiffness and muscle strain injury was identified. Elucidating the detailed mechanisms linking the identified locus to passive muscle stiffness may lead to the development of new strategies to prevent muscle strain injuries.

Humans↗

Correlation between muscle structure and filter characteristics of the muscle-joint system in three orthopteran insect species

In orthopteran insects, neural networks for joint control exhibit different characteristics due to behavioural specializations. We investigated whether these differences are generated purely by the neuronal networks, or whether characteristics of the muscles or joint architecture (muscle&shy;joint system) are also involved in these behavioural specializations. We compared the properties of the muscle system moving the femur&shy;tibia joint of the middle and hindleg of three species, Carausius morosus, Cuniculina impigra and Locusta migratoria. Four aspects were analysed for the tibial extensor muscle: (i) the frequency-dependence of motoneuronal activity in response to sinusoidal stimulation of the femoral chordotonal organ (fCO), (ii) the muscle structure, (iii) the innervation pattern of the muscle and (iv) the histochemical properties of the muscle fibres. These aspects were compared with the filter characteristics of the open-loop femur&shy;tibia control system and of the muscle&shy;joint system involved. Whereas in both phasmid species (Carausius morosus and Cuniculina impigra) the motoneuronal activity steadily increases with sinusoidal stimulation of the fCO in the frequency range 0.01&shy;5 Hz, in Locusta migratoria there is a decrease in motoneuronal activity between 0.01 and 0.3 Hz. The muscle structure is basically similar in all three species, as the number of singly innervated muscle fibres (supplied by the fast extensor tibiae motor neurone, FETi) decreases from proximal to distal. The number of triply innervated fibres supplied by the FETi, the slow extensor tibiae (SETi) and the common inhibitor 1 (CI1) is maximal in the middle of the muscle, and the number of dually innervated fibres (supplied by SETi, CI1) increases from proximal to distal. Differences between the locust and the two phasmid species exist in the distal portion of the muscle. The phasmid extensor tibiae muscle contains a morphologically distinct bundle of muscle fibres, not present in the locust, which is mostly dually innervated and which is larger in Cuniculina impigra. Similar results were obtained for the histochemical characterisation of the muscle fibres as revealed from their staining for myofibrillar ATPase activity. The number of histochemically identified fast fibres decreased from proximal to distal, while the number of slow fibres increased. In Carausius morosus and Locusta migratoria, the percentage of slow fibres increased by up to 60&shy;70 % at the distal end, while this increase was to almost 100 % in Cuniculina impigra. Apparently, the larger this distal region and the higher the percentage of slow, dually innervated fibres in it, the lower is the upper corner frequency (the stimulus frequency at which the joint control system produces a movement with 70 % of its maximal response amplitude) of the muscle&shy;joint system. In summary, it appears that the upper corner frequency of the open-loop system in Locusta migratoria (<0.05 Hz) results at least in part from properties of the neuronal joint control network, but in Carausius morosus (0.5&shy;1.0 Hz) and Cuniculina impigra (0.1&shy;0.2 Hz) it results from the upper corner frequency of the muscle&shy;joint system.

Journal Article↗

The splenius capitis muscle of the rat, architecture and histochemistry, afferent and efferent innervation as compared with that of the quadriceps muscle.

The splenius muscle of the rat was investigated with regard to its structure and innervation. The latter was compared with that of the quadriceps muscle. The results can be summarized as follows: The splenius muscles of both sides form a bipennate muscle plate connecting the occipital bone with the spinous process of the second thoracic vertebra. The lateral parts of both muscles are attached directly to this prominent bony process, whereas the medial parts end in a median raphe which forms a tendinous cranial extension of the second thoracic vertebra. This tendinous extension, showing no connection to the cervical vertebrae, serves also for the attachment of acromio-trapezius muscle fibers. The lateral part of the splenius muscle is divided into two parts by a tendinous intersection. The splenius muscle consists mainly of fast twitch fibers: 55% were characterized as IIB and 40% as IIA fibers by histochemical demonstration of myosin ATPase-activity. A high content of muscle spindles--57 spindles per gram of muscle tissue--was found. Comparing several aspects of the innervation of the splenius to that of the quadriceps muscle, the following results could be obtained: The ratio of motor end plate size to muscle fiber volume is significantly higher in the splenius than in the quadriceps muscle. As demonstrated by transganglionary HRP-transport, the main part of labeled splenius afferents to the spinal cord terminates in the central cervical nucleus. Quadriceps afferents, entering the lower thoracic and upper lumbar segments, mainly end in the area of Clarke's column. Several labeled fibers descend to the sixth lumbar and first sacral segments, where they terminate in the area of Stilling's nucleus. A group of primary afferents from both muscles--most probably III- and IV-afferents--projects to the dorsal laminae of the dorsal horn; terminals from the splenius are accumulated in the lateral parts of these laminae, whereas those of the quadriceps are more concentrated in the medial areas. Within the brain stem, most afferents from the splenius terminate in the external cuneate nucleus. Most of the quadriceps afferents course to the gracile nucleus. Terminals from both muscle nerves were found in the area of the spinal vestibular nucleus. In conclusion, the most conspicuous results were: 1) Besides the segmental projection to the dorsal horn there is an almost exclusive projection of splenius primary afferents to relay nuclei to the cerebellum. 2) The relatively high ratio: end plate size/muscle fiber volume, which is characteristic of finely adjusting muscles.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Differentiation of smooth muscle cells in chicken gizzard. Self-catalytic mechanism for the production of smooth muscle cells.

During development of the chicken gizzard, a thick layer of undifferentiated cells (mesenchymal cells) is constructed, and the cells differentiate into smooth muscle cells or connective tissues. We found that the differentiation of smooth muscle cells occurred first near the outer surface of the gizzard and the differentiated area spread to the inside of the gizzard. Therefore, we assumed that the differentiation of most of the smooth muscle cells in the gizzard is induced by differentiated smooth muscle itself. When undifferentiated cells from gizzard of 7-day-old embryo (Hamburger and Hamilton's stages 26-27) were cultured on a coverglass coated with extract of gizzard that contained differentiated smooth muscle cells, the cells attached to the coverglass and differentiated into smooth muscle cells. On the other hand, extract of gizzard from 7-day-old embryo did not induce the differentiation of smooth muscle cells, though it induced the attachment of cells. We found that activity for the differentiation of smooth muscle cells appeared when differentiated smooth muscle cells appeared in developing gizzard. Gizzard contained higher activity for the differentiation of smooth muscle cells than the other tissues. Transforming growth factor-beta (TGF-beta), which induces the differentiation of vascular smooth muscle cells, did not induce the differentiation of smooth muscle cells in gizzard, though extract of aorta induced the differentiation of smooth muscle cells in gizzard. The results obtained here support evidence that the differentiation of most of the smooth muscle cells in gizzard is induced by a self-catalytic mechanism in which differentiated smooth muscle itself induces the differentiation of smooth muscle cells.

Animals↗

Seven skeletal muscles rich in slow muscle fibers may function to sustain neutral position in the rodent hindlimb.

Skeletal muscles consist of slow-twitch and fast-twitch muscle fibers, which have distinct physiological and biochemical properties. The muscle fiber composition determines the contractile velocity and fatigability of a particular skeletal muscle. We analyzed the systemic distribution of slow muscle fibers in all rodent skeletal muscles by myosin ATPase staining and found that only seven hindlimb skeletal muscles were extremely rich in slow muscle fibers. These included the mouse piriformis (56.5%), gluteus minimus (35.7%), vastus intermedius (24.7%), quadratus femoris (69.9%), adductor brevis (44.3%), gracilis (24.6%), and soleus muscles (35.1%). In mice, the relative proportion of slow muscle fibers did not exceed 15% in skeletal muscles in other regions. The distribution of slow muscle fibers was well conserved in rats and rabbits. The soleus muscle is an important antigravity muscle in both rodents and humans; therefore, these skeletal muscles rich in slow muscle fibers might play an important role in sustaining neutral alignment of the lower extremity.

Amino Acid Sequence↗

Shoulder muscle reflex latencies under various levels of muscle contraction.

Previous research in relaxed muscles shows that muscle reflex latencies are too slow to protect the shoulder. However, during athletic activity when injury occurs, some level of shoulder muscle contraction typically exists. The purpose of the current study was to assess shoulder muscle reflex latencies under various levels of muscle contraction. Seventeen healthy subjects participated. A perturbation consisting of an external rotation collision force to the anterior forearm in a position of apprehension under various levels of muscle contraction (0%, 20%, and 50% of a maximum voluntary muscle contraction) was applied. Muscle reflex latencies were measured as the time from perturbation application to onset of muscle activity. Electromyography measured activity onset of the rotator cuff muscles and the primary humeral movers. During 0%, the latissimus dorsi muscle reflex latency was significantly slower than most other muscles. No difference existed between muscles in the 20% and 50% conditions. For the rotator cuff muscles, the reflex latencies significantly quickened for 20% and 50% compared with the relaxed state (0%). Overall, introducing muscle contraction significantly quickened muscle reflex latencies. These results provide clinicians with a better understanding of the role that these reflexes play in joint stability in a position of injury vulnerability like a position of apprehension.

Adult↗

Effects of SZ1677, a new non-depolarizing steroidal neuromuscular blocking drug, and rocuronium on two laryngeal muscles and the anterior tibial muscle in guinea pigs.

BACKGROUND: SZ1677 is a new neuromuscular blocking drug structurally related to rocuronium. We compared the effect of an ED(90) of SZ1677 (25 microg/kg) with that of rocuronium (100 microg/kg) in guinea pig laryngeal and peripheral muscles. METHODS: Electromyography was used to quantify neuromuscular blockade at the posterior cricoarytenoid muscle, the thyroarytenoid muscle and the anterior tibial muscle after SZ1677 (n = 10) and rocuronium (n = 9). RESULTS: Maximum neuromuscular blockade was similar after SZ1677 and rocuronium (83 +/- 11% vs. 89 +/- 11%; thyroarytenoid muscle: 91 +/- 8% vs. 97 +/- 3%; anterior tibial muscle: 91 +/- 15% vs. 96 +/- 3%, respectively). Onset time of neuromuscular blockade at the laryngeal muscles was similar for the two neuromuscular blocking drugs; it was shorter at the thyroarytenoid muscle (67 +/- 32 s vs. 42 +/- 40 s) than at the posterior cricoarytenoid muscle (101 +/- 26 s vs. 102 +/- 108 s). Onset time at the anterior tibial muscle was longer after SZ1677 (114 +/- 34 s) than after rocuronium (68 +/- 46 s); P < 0.05. Neuromuscular recovery was faster after SZ1677 (interval 25%-75%: posterior cricoarytenoid muscle: 222 +/- 66 s; thyroarytenoid muscle: 192 +/- 92 s; tibial muscle 149 +/- 55 s) than after rocuronium (450 +/- 148 and 464 +/- 183 s, 292 +/- 86 s, respectively); P < 0.05. CONCLUSIONS: In guinea pigs, SZ1677 offers a rapid onset of neuromuscular blockade at a laryngeal adductor muscle with a shorter duration than rocuronium. Regardless of the drug used, the course of neuromuscular blockade differs not only between peripheral muscles and the larynx but also between antagonistic laryngeal muscles. The differences seem to be species specific.

Androstanes↗

Ventilatory action of the hypaxial muscles of the lizard Iguana iguana: a function of slow muscle.

Patterns of muscle activity during lung ventilation, patterns of innervation and some contractile properties were measured in the hypaxial muscles of green iguanas. Electromyography shows that only four hypaxial muscles are involved in breathing. Expiration is produced by two deep hypaxial muscles, the transversalis and the retrahentes costarum. Inspiration is produced by the external and internal intercostal muscles. Although the two intercostal muscles are the main agonists of inspiration, neither is involved in expiration. This conflicts with the widely held notion that the different fibre orientations of the two intercostal muscles determine their ventilatory action. Several observations indicate that ventilation is produced by slow (i.e. nontwitch) fibres of these four muscles. First, electromyographic (EMG) activity recorded from these muscles during ventilation has an unusually low range of frequencies (less than 100 Hz). Such low-frequency signals have been suggested to be characteristic of muscle fibres that do not propagate action potentials (i.e. slow fibres). Second, during inspiration, EMG activity is restricted to he medical sides of the two intercostal muscles. Muscle fibres from this region have multiple motor endplates and exhibit tonic contraction when immersed in saline solutions of high potassium content. Like the intercostals, the transversalis and retrahentes costarum muscles also contain fibres with multiple motor endplates. Thus, although breathing is a phasic activity, it is produced by tonic (i.e. slow) muscle fibres. The intercostal muscles are also involved in postural and locomotor movements of the trunk. However, such movements employ twitch as well as slow fibres of the intercostal muscles.

Animals↗

Effect of one stretch a week applied to the immobilized soleus muscle on rat muscle fiber morphology.

We determined the effect of stretching applied once a week to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Twenty-six male Wistar rats weighing 269 +/- 26 g were divided into three groups. Group I, the left soleus was immobilized in the shortened position for 3 weeks; group II, the soleus was immobilized in the shortened position and stretched once a week for 3 weeks; group III, the soleus was submitted only to stretching once a week for 3 weeks. The medial part of the soleus muscle was frozen for histology and muscle fiber area evaluation and the lateral part was used for the determination of number and length of serial sarcomeres. Soleus muscle submitted only to immobilization showed a reduction in weight (44 +/- 6%, P = 0.002), in serial sarcomere number (23 +/- 15%) and in cross-sectional area of the fibers (37 +/- 31%, P < 0.001) compared to the contralateral muscles. The muscle that was immobilized and stretched showed less muscle fiber atrophy than the muscles only immobilized (P < 0.05). Surprisingly, in the muscles submitted only to stretching, fiber area was decreased compared to the contralateral muscle (2548 +/- 659 vs 2961 +/- 806 microm(2), respectively, P < 0.05). In conclusion, stretching applied once a week for 40 min to the soleus muscle immobilized in the shortened position was not sufficient to prevent the reduction of muscle weight and of serial sarcomere number, but provided significant protection against muscle fiber atrophy. In contrast, stretching normal muscles once a week caused a reduction in muscle fiber area.

Animals↗

Comparison of sensitivity of sodium currents to tetrodotoxin in equine muscle specimens with that in murine and human muscle specimens.

OBJECTIVE: To determine sensitivity of equine skeletal muscle to tetrodotoxin and compare that with sensitivity of murine and human skeletal muscles. SAMPLE POPULATION: Semimembranosus, vastus lateralis, triceps brachii, and masseter muscle specimens from 22 euthanatized horses, vastus lateralis muscle biopsy specimens from 25 clinically normal humans, and diaphragmatic muscle specimens from 6 mice. PROCEDURE: Electrically elicited twitch responses were measured in muscle specimens incubated in medium alone and with tetrodotoxin (100 nM, 400 nM, 1.6 microM for equine specimens and 100 nM, 200 nM, 400 nM, 800 nM, 1.6 microM for murine and human specimens). Percentages of tetrodotoxin-sensitive and -resistant sodium channels were determined and compared among muscles and species. RESULTS: 2 sodium channels with different sensitivities to tetrodotoxin were identified in equine muscle. One was blocked with 100 nM tetrodotoxin and the other was unaffected by tetrodotoxin at concentrations up to 1.6 microM. The only difference detected among the 4 equine muscles was that masseter muscle specimens had a higher percentage of tetrodotoxin-sensitive channels than triceps brachii muscle specimens. Tetrodotoxin-resistant sodium channels constituted 31 to 66% of the equine muscle twitch response, which was greater than that determined for normal human and murine muscle (< 5%). CONCLUSION AND CLINICAL RELEVANCE: Equine skeletal muscle contains a high percentage of tetrodotoxin-resistant sodium channels. The 4 equine muscles evaluated were more similar to each other than to murine and human muscles. Shifts in expression of sodium channel subtypes may play a role in the manifestation of certain myopathies.

Animals↗

Immunocytochemical electron microscopic study and Western blot analysis of caldesmon and calponin in striated muscle of the fruit fly Drosophila melanogaster and in several muscle cell types of the earthworm Eisenia foetida.

Caldesmon and calponin are two proteins that are characteristic of vertebrate smooth muscle. In invertebrates, caldesmon has only been studied in some molluscan muscles, and no previous references to calponin have been found. The aim of this paper was to investigate the presence and distribution of caldesmon and calponin in several invertebrate muscle cell types, classified according to their ultrastructural pattern: transversely striated muscle (flight muscle from Drosophila melanogaster), obliquely striated muscle (muscular body wall and inner muscular layer of the pseudoheart from the earthworm Eisenia foetida), and a muscle of doubtful classification which seems to be intermediate between smooth muscle and obliquely striated muscle (outer muscular layer of the pseudoheart, from E. foetida), using electron microscopy immunocytochemistry and Western blot analysis. Immunoreactions to both caldesmon and calponin were observed in the outer muscular layer cells from the earthworm pseudoheart but neither in the transversely striated muscle of D. melanogaster nor in the obliquely striated muscle from the earthworm. Present findings suggest that caldesmon- and calponin-like proteins are also present in invertebrate muscle cells, but only in those that are ultrastructurally similar to the vertebrate smooth muscle cells. Since discrepancies in the classification of some invertebrate muscles are common in the literature, the use of distinctive markers, such as troponin, caldesmon and calponin may improve our understanding of the nature and properties of many invertebrate muscles showing an ultrastructural pattern that does not resemble any of the classic muscle types.

Animals↗