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Effects of experimental muscle pain on muscle activity and co-ordination during static and dynamic motor function.

The relation between muscle pain, muscle activity, and muscle co-ordination is still controversial. The present human study investigates the influence of experimental muscle pain on resting, static, and dynamic muscle activity. In the resting and static experiments, the electromyography (EMG) activity and the contraction force of m. tibialis anterior were assessed before and after injection of 0.5 ml hypertonic saline (5%) into the same muscle. In the dynamic experiment, injections of 0.5 ml hypertonic saline (5%) were performed into either m. tibialis anterior (TA) or m. gastrocnemius (GA) and the muscle activity and co-ordination were investigated during gait on a treadmill by EMG recordings from m. TA and m. GA. At rest no evidence of EMG hyperactivity was found during muscle pain. The maximal voluntary contraction (MVC) during muscle pain was significantly lower than the control condition (P < 0.05). During a static contraction at 80% of the pre-pain MVC muscle pain caused a significant reduction in endurance time (P < 0.043). During dynamic contractions, muscle pain resulted in a significant decrease of the EMG activity in the muscle, agonistic to the painful muscle (P < 0.05), and a significant increase of the EMG activity of the muscle, antagonistic to the painful muscle (P < 0.05). Muscle pain seems to cause a general protection of painful muscles during both static and dynamic contractions. The increased EMG activity of the muscle antagonistic to the painful muscle is probably a functional adaptation of muscle co-ordination in order to limit movements. Modulation of muscle activity by muscle pain could be controlled via inhibition of muscles agonistic to the movement and/or excitation of muscles antagonistic to the movement. The present results are in accordance with the pain-adaptation model (Lund, J.P., Stohler, C.S. and Widmer, C.G. In: H. Vaerøy and H. Merskey (Eds.), Progress in Fibromyalgia and Myofascial Pain. Elsevier, Amsterdam, 1993, pp. 311-327.) which predicts increased activity of antagonistic muscle and decreased activity of agonistic muscle during experimental and clinical muscle pain.

Adult↗

Stimulation of chemosensitive afferents from multifidus muscle does not sensitize multifidus muscle spindles to vertebral loads in the lumbar spine of the cat.

STUDY DESIGN: Electrophysiologic recordings from muscle spindle afferents innervating the lumbar multifidus muscle of the cat while loading the L6 vertebra at its spinous process and while exposing the segmentally adjacent lumbar multifidus muscles to algesic and inflammatory mediators. OBJECTIVES: The purpose of this study was to investigate a possible mechanism underlying muscle spasm and pain in the lumbar spine. The hypothesis was tested that stimulation of chemosensitive afferents with receptive endings in the paraspinal muscle increases the discharge of paraspinal muscle spindle afferents during loading of a lumbar vertebra. The presence of such a phenomenon would provide a mechanism by which pain or inflammation could alter segmental lumbar biomechanics and contribute to lumbar spine dysfunction. SUMMARY OF BACKGROUND DATA: Muscle pain, tenderness, and altered muscle tone are often associated with musculoskeletal disorders. The literature suggests that stimulation of Group III and IV muscle afferents sensitive to algesic or inflammatory metabolites increases the stretch sensitivity of muscle spindles via a reflex pathway involving gamma-motoneurons. The reflex increase in muscle spindle activity, in turn, reflexly increases the excitability of alpha-motoneurons leading to enhanced muscle tone and the further accumulation of muscle metabolites and subsequent pain. Studies in the cervical spine support this hypothesis. It has not been investigated in the lumbar spine. METHODS: Single unit activity from muscle spindles in the L6 multifidus muscle were recorded from the cut peripheral end of the L6 dorsal root in alpha-chloralose-anesthetized cats and in decerebrate unanesthetized cats. The L6 vertebra was loaded at its spinous process using a force-feedback motor. Ramp and hold loads were delivered at 25%, 50%, 75%, and 100% body weight. Chemosensitive afferents in the L5 and L7 multifidus muscle were stimulated by bathing (subfascial injection) or infiltrating (intramuscular injection) the L5 and L7 multifidus muscles with bradykinin or capsaicin. RESULTS: Loading the L6 vertebra stimulated muscle spindles in the L6 multifidus muscle. Neither the saline volume control nor bradykinin nor capsaicin injected subfascially or intramuscularly affected the response of L6 multifidus muscle spindles to ramp and hold vertebral loads in the alpha-chloralose-anesthetized cat. In addition, neither saline nor bradykinin nor capsaicin injected intramuscularly affected the activity of L6 multifidus muscle spindles to ramp and hold vertebral loads in the unanesthetized decerebrate cat. CONCLUSIONS: These results indicate that stimulation of small diameter muscle afferents in a deep muscle of the lumbar spine does not sensitize muscle spindles to vertebral loads. These data do not support the hypothesis that fusimotor reflexes evoked by chemosensitive muscle afferents contribute to muscle spasm or to changes in muscle tone in the lumbar spine. In addition, the present results do not provide evidence for the pain-spasm-pain cycle in the lumbar spine.

Action Potentials↗

The period of latency before a muscle receptor generates an action potential as a response to a muscle stretch.

Six primary (Ia) and seven secondary (II) muscle spindle afferents and eight Golgi tendon organ afferents (Ib) from the tibial anterior muscle of the cat, recorded at the dorsal roots, were subjected to a sinusoidal stretch of the host muscle, the frequency of which increased linearly from 2 to 80 Hz over four different lengths of time. Both the amplitude of the sinusoidal stretch and the prestretch of the muscle were varied. The phase of the action potentials was determined. The phase of the action potential, driven 1:1, increased linearly with frequency. From the gradient of the phase of this action potential the muscle-muscle receptor latency was determined, i.e., the period of latency between the stretch of the muscle and the occurrence of the action potential at the muscle nerve where it enters the muscle. The muscle-muscle receptor latency had values lying between 3 and 8 ms: it was dependent on the experimental parameters and became shorter as the conduction velocity of the afferent fiber increased. In three experiments the muscle latency was determined, i.e., the period of latency before the stretch was transferred from the tendon of the muscle to the proximal third of the muscle belly. The muscle was stretched sinusoidally under the same varying parameters as given above. The length changes occurring in the proximal third of the muscle were measured with a piezo element. The muscle latency was determined from the slope of the phase of the zero points of the sinusoidal piezo length changes; the phase increases linearly with frequency. The muscle latency had values lying between 6 and 15 ms: it was dependent on the experimental parameters. The muscle spindle latency, i.e., the period of latency between the stretch of the polar parts of the intrafusal muscle fibers and the recording of the action potentials from the spindle nerve near the spindle capsule, was determined from 5 Ia fibers and 1 II fiber of isolated muscle spindles. The isolated muscle spindle was stretched under the same varying parameters as given above. The muscle spindle latency was determined from the slope of the phase of the phase-locked action potential. The muscle spindle latency as measured by our method proved to be 0 ms. The latencies of the three elements and their dependence on the experimental parameters are discussed in the light of the transfer properties of the muscle and the muscle receptors.

Action Potentials↗

Muscle trophic factor: I. Assay of a muscle trophic factor by measurement of muscle cell nuclei.

A method for assaying a skeletal muscle cell trophic factor in chicken serum is described. The number of myoblast and myotube nuclei was considered to reflect the rate of myoblast multiplication and was thus adopted as an index for the activity of the trophic factor. In a 35-mm Falcon plastic Petri dish, 2.5 x 10(40) to 10(5) myoblasts were cultured. The culture medium consisted of 85% Eagle's minimum essential medium and 15% horse serum. The assay medium was composed of 2.4 ml of this culture medium and 0.1 ml of partially purified trophic factor or appropriately diluted chicken serum. Four dishes were prepared for each sample under identical conditions. On day 4 of incubation, cells were fixed, stained and observed under a microscope, and the number of nuclei in 20 fields was counted. The mean number of nuclei obtained from the four dishes was used to denote a single point. A three-point common-zero method of slope-ratio assay was used to calculate activity. The activity of the trophic factor was expressed with reference to the effect of the control serum on myoblast multiplication.

Animals↗

Smooth muscle-type myosin heavy chain isoforms in bovine smooth muscle and non-muscle tissues.

The distribution of smooth muscle (SM)-type myosin heavy chain isoforms in several bovine muscular and non-muscular (NM) tissues was evaluated by immunofluorescence tests using monoclonal antibodies SM-E7, reactive with 204 (SM1) and 200 (SM2) kDa isoforms, and SM-F11, specific for SM2 isoform. SM-E7 reacted equally with vascular, respiratory and intestinal SM tissues, whereas SM-F11 stained heterogeneously SM cells in the various muscular systems examined and in some peculiar tissues was unreactive (perisinusoidal cells of hepatic lobule, pulmonary interstitial cells and intestinal muscularis mucosae) or uniquely reactive (nerve cells). On the whole, our findings indicate that SM1 and SM2 isoforms are unequally distributed at the cellular level in various SM and NM tissues and support previous results obtained with tissue extracts and electrophoretic procedures.

Animals↗

Heterogeneity among muscle precursor cells in adult skeletal muscles with differing regenerative capacities.

Skeletal muscle has a remarkable capacity to regenerate after injury, although studies of muscle regeneration have heretofore been limited almost exclusively to limb musculature. Muscle precursor cells in skeletal muscle are responsible for the repair of damaged muscle. Heterogeneity exists in the growth and differentiation properties of muscle precursor cell (myoblast) populations throughout limb development but whether the muscle precursor cells differ among adult skeletal muscles is unknown. Such heterogeneity among myoblasts in the adult may give rise to skeletal muscles with different regenerative capacities. Here we compare the regenerative response of a masticatory muscle, the masseter, to that of limb muscles. After exogenous trauma (freeze or crush injuries), masseter muscle regenerated much less effectively than limb muscle. In limb muscle, normal architecture was restored 12 days after injury, whereas in masseter muscle, minimal regeneration occurred during the same time period. Indeed, at late time points, masseter muscles exhibited increased fibrous connective tissue in the region of damage, evidence of ineffective muscle regeneration. Similarly, in response to endogenous muscle injury due to a muscular dystrophy, widespread evidence of impaired regeneration was present in masseter muscle but not in limb muscle. To explore the cellular basis of these different regenerative capacities, we analyzed the myoblast populations of limb and masseter muscles both in vivo and in vitro. From in vivo analyses, the number of myoblasts in regenerating muscle was less in masseter compared with limb muscle. Assessment of population growth in vitro indicated that masseter myoblasts grow more slowly than limb myoblasts under identical conditions. We conclude that the impaired regeneration in masseter muscles is due to differences in the intrinsic myoblast populations compared to limb muscles.

Animals↗

Effects of vecuronium and rocuronium in antagonistic laryngeal muscles and the anterior tibial muscle in the cat.

BACKGROUND: Adequate vocal cord paralysis and full recovery of laryngeal muscle function are important when muscle relaxants are used perioperatively. This study was designed to compare the effects of vecuronium and rocuronium at the vocal cord abductor and adductor muscles and the anterior tibial muscle in cats. METHODS: Twelve adult cats were studied under pentobarbitone-N2O/O2-anesthesia. After supramaximal electrical stimulation of the peroneal nerve and the recurrent laryngeal nerve (0.1 Hz and intermittent train-of-four) evoked electromyographic responses were obtained from the anterior tibial muscle, the posterior cricoarytenoid muscle (vocal cord abductor) and two vocal cord adductor muscles, the lateral cricoarytenoid and the vocal muscle. Six cats received bolus doses of increasing size of vecuronium (ED90 22.5 microg x kg(-1)) and six cats rocuronium (ED90 90 microg x kg(-1)). RESULTS: Equipotent doses of vecuronium and rocuronium caused a similar degree of paralysis in all muscles (vecuronium ED90: 70% blockade at the posterior cricoarytenoid, 83% at the lateral cricoarytenoid, 84% at the vocal muscle and 90% at the anterior tibial muscle; rocuronium ED90: 71% at the posterior cricoarytenoid, 67% at the lateral cricoarytenoid, 78% at the vocal muscle and 90% at the anterior tibial muscle; vecuronium 2 x ED90: 93% blockade at the posterior cricoarytenoid, 95% at the lateral cricoarytenoid, 97% at the vocal muscle and 99% at the anterior tibial muscle; rocuronium 2 x ED90: 89% blockade at the posterior and lateral cricoarytenoid, 93% at the vocal muscle and 100% at the anterior tibial muscle). Onset time was significantly shorter at the posterior cricoarytenoid muscle (290 s) compared to the lateral cricoarytenoid muscle (400 s) after vecuronium ED90 and to the vocal muscle (150 s versus 210 s) after rocuronium ED90. Compared to the anterior tibial muscle (interval 25-75%: 6.5 min after vecuronium 2 x ED90 and 3.3 min after rocuronium 2 x ED90 and to the posterior cricoarytenoid muscle (interval 25-75%: 7 min after vecuronium 2 x ED90 and 4.3 min after rocuronium 2 x ED90), recovery of laryngeal adductor muscle function was markedly delayed with both neuromuscular blocking drugs (interval 25-75% at the lateral cricoarytenoid and vocal muscle: 14 min and 15.8 min after vecuronium 2 x ED90 and 10.3 min and 11.6 min after rocuronium 2 x ED90 respectively). CONCLUSION: In cats, the time course of neuromuscular blockade after vecuronium and rocuronium differs in antagonistic laryngeal muscles. The protective laryngeal function of glottis closure recovers later than vocal cord abduction after both vecuronium and rocuronium.

Androstanols↗

Three-dimensional architecture of the intrinsic tongue muscles, particularly the longitudinal muscle, by the chemical-maceration method.

Muscle bundles of the transverse and vertical muscles of the tongue become flat when they enter the longitudinal muscle layers of the tongue, where they form a tunnel-like structure that surrounds the longitudinal muscle of the tongue. However, the three-dimensional architecture of longitudinal muscle fibers of the tongue has not been clarified. In the present study, we evaluated the function of the intrinsic muscles of the tongue by studying the three-dimensional architecture of the longitudinal muscle. Muscle bundles of the longitudinal muscle of the anterior part of a rabbit's tongue were exposed by the chemical-maceration and modified chemical-maceration methods and examined by scanning electron microscopy. In the longitudinal muscle of the tongue, muscle bundles running in the anteroposterior direction were arranged at regular intervals. These muscle bundles bifurcated or ramified at a sharp angle at each level from the superficial layer to the deep layer and joined or fused with adjacent muscle bundles. In addition, these ramified muscle bundles ran obliquely into shallower or deeper layers of the muscle, as well as in the same plane. Consequently, the longitudinal muscle of the tongue as a whole had a three-dimensional mesh-like structure. The transverse and vertical muscles of the tongue entered this mesh-like structure of muscle bundles of the longitudinal muscle as flat muscle bundles. The transverse and vertical muscles showed no ramification in the center of the tongue, where there is no longitudinal muscle. These results suggest that the three intrinsic muscles of the tongue are interlaced with one another and are bound tightly in the longitudinal muscle. This structure may enable the dorsum of the tongue to harden for pressing food during mastication and shifting the food posteriorly for swallowing.

Animals↗

Distribution, density, and structure of muscle spindles in the vastus intermedius and the peroneus longus muscles of sheep.

Muscle spindles are not always distributed more in postural muscles with many slow-twitch-oxidative (SO) myofibers than in locomotory muscles with few SO myofibers. The purpose of present study was to examine the distribution, density, and structure of muscle spindles in the vastus intermedius muscle: an antigravity muscle and the peroneus longus muscle: a locomotory muscle in the sheep. Muscle spindles were reconstructed from serial sections at 300 microns intervals throughout the muscles. Myofiber types were classified into SO, fast-twitch-oxidative-glycolytic, and fast-twitch-glycolytic myofibers by differences in histochemical reactivity. No significant difference in the density of muscle spindles (DMS) existed between the vastus intermedius (DMS: 5.3) and peroneus longus (DMS: 5.7) muscles. The muscle spindles were distributed more in the distal portion than in the proximal portion of the vastus intermedius muscle. The muscle spindles were distributed in the proximal and middle portion but hardly in the distal portion of the peroneus longus muscle. Muscle spindles were classified into simple, tandem, and compound muscle spindles. Most of the muscle spindles were the simple type. The differences in size of the muscle spindle and numbers of the intrafusal myofibers were not significant between the two muscles. The results show that the density and structure of the muscle spindles do not differ between the postural and locomotory muscles in the sheep.

Animals↗

Shortening of muscle fibres during stretch of the active cat medial gastrocnemius muscle: the role of tendon compliance.

1. The length of muscle fibres in the medial gastrocnemius (MG) muscle of the anaesthetized cat was measured using ultrasound techniques. During the course of 'isometric' contractions, the muscle fibres shortened by stretching the compliant tendons, until the muscle fibres could no longer produce enough force to stretch the tendons further. At optimal muscle length (Lo) the maximal shortening of muscle fibres was 28%. 2. At muscle lengths much longer than Lo, 'isometric' contractions produced a slow shortening of the muscle fibres as the tendons were stretched and this resulted in a slow rise in tension. This phenomenon, usually referred to as 'creep', is due to low power at long muscle fibre length. This study shows that the series compliance present in the tendons is the major contributor to 'creep' in the cat MG muscle. As the tendons stretched during the course of the contraction, the average sarcomere length became shorter providing greater filament overlap and increasing power. 3. Slow to medium speed stretches applied shortly after the onset of contraction, as occurs in cat MG during walking and trotting, were entirely taken up in the tendons and the muscle fibres actually shortened throughout the imposed muscle stretch. 4. When early stretches were applied at muscle lengths longer than Lo, stretch of the muscle resulted in a peak force that was less than if the stretch had not been applied. This was the reverse of the situation for stretches at lengths less than Lo. When stretch was applied after attaining peak force, the force was greatly enhanced and the muscle fibres were also stretched. 5. Using the same techniques in a freely walking cat, the muscle fibres shortened by 1.0 +/- 0.3 mm during the stance phase of the step-cycle when the muscle was being stretched, in 198 consecutive step-cycles. 6. The tendons act as a mechanical buffer to protect muscle fibres from damage during eccentric contractions. 7. Since stretches of the MG muscle are not faithfully imposed on the muscle fibres, studies of muscle spindle function during locomotion need to take into consideration these effects of tendon compliance. The dominant view, when the foot lands on the ground during normal locomotion, is that muscle spindles are stretched along with the muscle resulting in reflex enhancement of contractile force. This study shows that the muscle fibres do not stretch under these circumstances, except at high speeds of locomotion when the stretch rate is also high.

Animals↗

Characterization of several invertebrate muscle cell types: a comparison with vertebrate muscles.

Ultrastructural classification of invertebrate muscles is complex and not always clear. The aim of the present paper was to establish some criteria that might be useful for classification of invertebrate muscles and for a better understanding of the differences between them. The procedures used were: (1) immunochemical evaluation of those proteins that differentiated striated from smooth muscle (troponin, caldesmon, and calponin), and (2) calculations of several myofilament parameters to establish differences among muscles. The muscles studied were: striated muscles from the rat, Drosophila, the crab Callinectes, and the snail Helix (heart); obliquely striated muscles from the earthworm Eisenia foetida and Helix (mouth); and smooth muscles from the rat, and Helix (retractor, body wall, and intestinal wall). Immunochemical studies revealed that troponin was only present in the striated muscles and the obliquely striated muscle from Eisenia, whereas caldesmon and calponin were only present in the smooth muscles and the obliquely striated muscle from Helix. The highest thick filament/thin filament volume ratio was found in the striated muscles, followed by the obliquely striated muscles, and the smooth muscles. This suggests the order in which the contraction strength decreases. The myofilament length is inversely related to the contraction speed, which was higher in the striated muscles than in the obliquely striated muscles. In vertebrates, the smooth muscle seems to be less rapid than the striated muscle because their myofilaments are longer. This assertion cannot be generalized for invertebrate smooth muscle, because myofilament lengths vary widely in both striated and smooth muscles. In smooth muscles, the presence of apparently unordered electron-dense bodies instead of ordered Z lines and the absence of true sarcomeres permit a certain overlapping of thin filaments increasing the range of shortening.

Actin Cytoskeleton↗

Interrelations of myogenic response, progressive atrophy of muscle fibers, and cell death in denervated skeletal muscle.

Little is known concerning the time-course and structural dynamics of reactivation of compensatory myogenesis in denervated muscle, its initiating cellular mechanisms, and the relationship between this process and the progression of postdenervation atrophy. The purpose of this study was to investigate the interrelations between temporal and spatial patterns of the myogenic response in denervated muscle and progressive atrophy of muscle fibers. Another objective was to study whether reactivation of myogenesis correlates with destabilization of the differentiated state and death of denervated muscle cells. It has remained unclear whether muscle fiber atrophy was the primary factor activating the myogenic response, what levels of cellular atrophy were associated with its activation, and whether the initiation and intensity of myogenesis depended on the local and individual heterogeneity of atrophic changes among fibers. For this reason, our objective was also to identify the levels of atrophic and degenerative changes in denervated muscle fibers that are correlated with activation of the myogenic response. We found that the reactivation of myogenesis in the tibialis anterior and extensor digitorum longus muscles of the rat starts between days 10-21 following nerve transection, before atrophy has attained advanced level, long before dead cells are found in the tissue. Formation of new muscle fibers reaches its maximum between 2 and 4 months following denervation and gradually decreases with progressive postdenervation atrophy. The myogenic response is biphasic and includes two distinct processes. The first process resembles the formation of secondary and tertiary generations of myotubes during normal muscle development and dominates during the first 2 months of denervation. During this period, activated satellite cells form new myotubes on live differentiated muscle fibers. Most of the daughter myotubes in 1- and 2-month denervated muscle develop on the surface of fast type parent muscle fibers, and some of the newly formed muscle fibers express slow myosin. Some fast type parent fibers are weakly or, more rarely, moderately immunopositive for embryonic isomyosin. This indicates that reactivation of myogenesis may also depend on the fiber type. The level of atrophy, destabilization of the differentiated myofiber phenotype, and degenerative changes of individual fibers in denervated muscle are very heterogeneous. The myogenic response of the first type is associated predominantly with fibers of average and higher than average levels of atrophy. Muscle cells that undergo a lesser degree of atrophy also form daughter fibers, although with a lower incidence. We did not find any correlation between the size of newly formed fibers and the level of atrophy of parent fibers. The topographical distribution of new myotubes both in the peripheral and central areas of the mid-belly equatorial sections at the early stages following nerve transection indicates that myogenesis of the first type represents a systemic reaction of muscle to the loss of neural control. These data indicate that activation of the myogenic response does not depend on cell death and degenerative processes per se. The second type of myogenesis is a typical regenerative reaction that occurs mainly within the spaces surrounded by the basal laminae of dead muscle fibers. Myocytes of different sizes are susceptible to degeneration and death, which indicates that cell death in denervated muscle does not correlate with levels of muscle cell atrophy. The regenerative process frequently results in development of abnormal muscle cells that branch or form small clusters. Replacement of lost fibers becomes activated between 2 and 4 months following nerve transection, i.e., mainly at advanced stages of postdenervation atrophy, when cell death becomes a contributing factor of the atrophic process. In long-term denervated muscle, the first and second types of myogenesisoccur concurrently, and the topographical distribution of the myogenic response becomes more heterogeneous than during the first weeks following denervation. Thus, our data demonstrate differential temporal and spatial expression of two patterns of myogenesis in denervated muscle that appear to be controlled by different regulatory mechanisms during the postdenervation period. (c) 2001 Wiley-Liss, Inc.

Animals↗

Electrical coupling of circular muscle to longitudinal muscle and interstitial cells of Cajal in canine colon.

1. Electrical communication between circular muscle, longitudinal muscle and interstitial cells of Cajal (ICC) was investigated; the hypothesis was tested that the resting membrane potential (RMP) gradient in the circular muscle of canine colon is caused by electrical coupling to neighbouring cells. 2. Isolated longitudinal muscle exhibited spike-like action potentials at a RMP of -45 mV with a frequency and amplitude of 20 cycles/min and 12 mV, respectively. 3. The circular muscle (CM), devoid of longitudinal muscle, myenteric plexus and submuscular ICC-smooth-muscle network, was electrically quiescent at a uniform RMP of -62 mV across the entire circular muscle layer. 4. Preparations consisting of only the submuscular ICC network and a few adjacent layers of circular muscle cells exhibited slow wave-type action potentials at a RMP of about -80 mV. 5. In ICC-CM preparations, consisting of the submuscular ICC network and circular muscle, a RMP gradient of 10 mV was observed near the submucosal border, whereas the RMP was constant at -62 mV in the myenteric half of the circular muscle. 6. In full thickness (FT) preparations, a RMP gradient of 23 mV was observed. The RMP decreased gradually from -71 mV at the submucosal border to -48 mV at the myenteric border of the circular muscle. 7. Coupling of longitudinal muscle to circular muscle caused circular muscle cells at the myenteric surface to depolarize by 14 mV and longitudinal muscle cells to hyperpolarize by 3 mV. 8. In the ICC-CM preparations, the slow wave amplitudes did not decay exponentially away from the ICC network indicating that slow waves propagated actively into the circular muscle; in the FT preparations there was an apparent exponential decay but this was due to the RMP gradient. 9. Spike-like action potentials (SLAPs) superimposed on the plateau phase of slow waves did not decay exponentially away from the myenteric border suggesting that SLAPs were generated within the circular muscle layer. 10. In summary, circular muscle cells possess a uniform intrinsic RMP of -62 mV. The RMP gradient in situ is caused by electrical coupling of circular muscle cells to longitudinal muscle cells and the submuscular network of ICC. In situ, slow wave-type action potentials propagate actively into the circular muscle layer, and, dependent on the level of excitation, circular muscle cells actively generate spikes.

Action Potentials↗

Mechanisms causing effects of muscle position on proximo-distal muscle force differences in extra-muscular myofascial force transmission.

Certain recent studies showed that extra-muscular myofascial force transmission affects the length-force characteristics of rat extensor digitorium longus (EDL) muscle significantly after distal or proximal lengthening. This suggested that the relative position of a muscle with respect to its surrounding connective tissues is a co-determinant of muscle force in addition to muscle length, and indicated major effects on muscular mechanics. The specific goal of the present study is to investigate such effects by studying: (1) distributions of lengths of sarcomeres within muscle fibres and (2) the relative contributions of muscle fibres and the extra-cellular matrix to muscle total force, using a finite element model. The length of the muscle modelled was kept constant at a high and at a low muscle length whereas the relative position of the muscle was altered exclusively. For both muscle lengths, the forces exerted at distal and proximal tendons were unequal at almost all muscle relative positions. The proximo-distal force difference was enhanced as the muscle was repositioned away from its reference position. This confirmed the role of relative position of a muscle as a co-determinant of muscle force. At higher muscle lengths, distributions of lengths of sarcomeres arranged in series within muscle fibres were substantial. The force transmitted by the muscles' extra-cellular matrix comprised a sizable part of muscle total force. At lower muscle lengths distribution of sarcomere lengths was relatively limited indicating that the extra-cellular matrix is bearing the extra-muscular force. However, minor sarcomere length changes were shown to accumulate to sizable effects on the summed forces exerted by the muscle fibres. In addition, the extra-muscular load was shown to manipulate the force exerted by the extra-cellular matrix. We conclude that the relative position of a muscle has substantial effects on intra-muscular mechanics and the importance of the role of the extra-cellular matrix in determining the proximo-distal force differences is comparable to that of the intra-cellular domain.

Animals↗

Fiber-type composition and fiber size of the human cricopharyngeal muscle and the pharyngeal constrictor muscle.

BACKGROUND: Despite a similar density of nicotinic acetylcholine receptors, the upper esophageal sphincter is sensitive to partial neuromuscular block, whereas the pharyngeal constrictor muscle is more resistant. In order to postulate possible mechanisms behind this difference in pharmacological response, basic knowledge of morphological and physiological features of these muscles is needed. The aim of this study was to compare the muscle fiber-type composition, the size and the morphology of the muscle fibers of the cricopharyngeal muscle, the main component of the upper esophageal sphincter, with that of the pharyngeal constrictor muscle. METHODS: Muscle specimens were obtained from five patients undergoing surgery with laryngectomy. Muscle fiber type was determined by myosin heavy chain immunohistochemistry and the muscle fiber cross-sectional area was measured for each fiber type by planimetry. Morphology of muscle fibers was evaluated by histochemistry. RESULTS: The muscle fiber cross-sectional area was generally smaller in the cricopharyngeal muscle compared with the pharyngeal constrictor muscle (P < 0.001). The composition of fiber types showed a large interindividual variability with no distinct difference between the studied muscles. Aberrant histological features were common in both the cricopharyngeal muscle and the pharyngeal constrictor muscle. CONCLUSION: The main morphological difference between the neuromuscular blocking agents sensitive cricopharyngeal muscle and the more resistant pharyngeal constrictor muscle is a uniformly smaller size of contributing fiber types in the cricopharyngeal muscle than in the pharyngeal constrictor muscle. The muscle fiber-type composition does not differ between the two studied muscles.

Adult↗

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

BACKGROUND: There is little information about troponin in invertebrate muscles, and no previous references to this protein in annelid muscles have been found. The aim of this paper was to study the presence and distribution of troponin in different muscle cell types from the earthworm Eisenia foetida (the muscular body wall, and the inner and outer muscular layer of the pseudoheart). These results were compared with those obtained in the transversely striated muscle of Drosophila melanogaster and in skeletal and smooth muscles of the mouse. METHODS: Immunocytochemical electron microscopic study and Western blot analysis using anti-TnT antibodies were employed in this study. RESULTS: Troponin immunoreaction was detected in the mouse skeletal muscle, the fly flight muscle, and earthworm obliquely striated muscles (body wall musculature and inner muscular layer of the pseudoheart). Immunolabeling for TnT in all these muscle cells appeared in moderate amounts at any point along the sarcomere length, except for the central zone of the A band (H band). This suggests that troponin molecules were located along the thin filaments. The density of immunogold particles was similar in the three muscles, and thus the amount of troponin in each muscle type was proportional to the number and length of actin filaments in each. Troponin was found in neither the mouse smooth muscle nor the outer muscular layer of the earthworm pseudoheart. The latter muscle showed an ultrastructural pattern that was intermediate between obliquely striated and smooth muscle. The estimated molecular weight for TnT in the earthworm was 55 kDa; this is higher than the weight of this protein in the mouse skeletal muscle (40 kDa) but similar to that of the D. melanogaster muscle (52 kDa). CONCLUSIONS: Troponin is present in both types of striated muscle (transversely striated and obliquely striated) of the earthworm with a distribution that is very similar to that observed in the mammalian striated muscle. As in vertebrates, troponin is absent in the smooth muscle of the earthworm. Discrepancies in the classification of some invertebrate muscles are common in the literature, and the use of distinctive markers, such as troponin, 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↗

The pelvic floor muscles: muscle thickness in healthy and urinary-incontinent women measured by perineal ultrasonography with reference to the effect of pelvic floor training. Estrogen receptor studies.

Maintenance of urinary continence is multifactorial and depends mainly on detrusor control and urethral closure function. The closure forces can be categorized as permanent closure forces active at rest, and adjunctive closure forces active during physical activities. The efficiency of these forces depends on the structural components in the urethral wall, the position of the bladder neck and proximal urethra, the periurethral striated muscles, and the pelvic floor muscles. By means of pudendal blockade and simultaneous recordings of pressure and cross-sectional area in the urethra, it has been demonstrated that the striated periurethral muscles and the pelvic floor muscles are of paramount importance for the closure function. This emphasizes the importance of well-functioning pelvic floor muscles to obtain continence, and probably explains the rationale for the effect of pelvic floor training in treating urinary incontinence. This study presents a review of the literature on female urinary incontinence, continence mechanisms, pelvic floor muscles, and pelvic floor training. Furthermore, a review of the literature on estrogen receptors in the pelvic floor muscles is given. Perineal ultrasonography, a method for visualization and measurement of thickness of the pelvic floor muscle, was developed and evaluated. This method was used to gain information on the thickness of the pelvic floor muscles in younger physiotherapists, healthy women, and women suffering from urinary incontinence, and to evaluate the effect of pelvic floor training. Additionally, a study of the Pelvic floor muscles was performed to assess the presence of estrogen receptors. Muscle thickness seems to decrease with age. In women over age 60 years, a significantly thinner pelvic floor muscle was found compared to younger women. The muscle increment during contraction decreased significantly with age, probably reflecting a stronger pelvic floor or a better awareness of pelvic floor function in the younger women. Incontinent women had a thinner pelvic floor muscle compared to healthy women. Hypertrophy of the muscles was demonstrated in urinary-incontinent women after pelvic floor training, and the difference in thickness of the muscles in these women before training compared to healthy women was eliminated by training. pelvic floor training reduced the use of incontinence appliances and urinary leakage both in stress and urge-incontinent women. Subjectively, 60% of the women gained a positive effect of the training. In spite of the fact that training increased muscle thickness and the increment of muscle thickness during contraction, no correlation between these parameters and subjective improvement or reduced urine loss in the pad weighing test could be demonstrated. Training may strengthen the pelvic floor without effect on the multifactorial continence mechanism in cases where urinary incontinence is caused by destruction of the urethral attachment to the surrounding tissue. No estrogen receptors were found in the nuclei of striated muscle cells in biopsies from levator ani muscles, using an immunohistochemical technique. Thus, the effect of estrogen treatment on the striated pelvic floor muscles is doubtful. A possible effect of estrogen treatment of urinary incontinence must be mediated via other structures than the pelvic floor muscles.

Exercise Therapy↗

PDGF-receptor concentration is elevated in regenerative muscle fibers in dystrophin-deficient muscle.

Dystrophin-deficient muscle undergoes sudden, postnatal onset of muscle necrosis that is either progressive, as in Duchenne muscular dystrophy, or successfully arrested and followed by regeneration, as in most muscles of mdx mice. The mechanisms regulating regeneration in mdx muscle are unknown, although the possibility that there is renewed expression of genes regulating embryonic muscle cell proliferation and differentiation may provide testable hypotheses. Here, we examine the possibility that necrotic and regenerating mdx muscles exhibit renewed or increased expression of PDGF-receptors. PDGF-binding to receptors on muscle has been shown previously to be associated with myogenic cell proliferation and delay of muscle differentiation. We find that PDGF-receptors are present in 4-week-old mdx mice in muscles that undergo brief, reversible necrosis (hindlimb muscles) or progressive necrosis (diaphragm), as well as in 4-week-old control mouse muscles. Immunoblots indicate that the concentrations of PDGF-receptors in 4-week-old dystrophic (necrotic) and control muscles are similar. Prenecrotic, dystrophic fibers and control fibers possess some cell surface labeling of fibers treated with anti-PDGF-receptor and viewed by indirect immunofluorescence. Necrotic fibers in dystrophic muscle show cytoplasmic labeling for PDGF-receptors and labeling of perinuclear regions at the muscle cell surface. Adult dystrophic muscle displays higher concentrations of PDGF-receptor in both regenerated muscle (hindlimb) and progressively necrotic muscle (diaphragm) than found in controls. Anti-PDGF-receptor labeling of regenerated, dystrophic muscle is observed primarily in granules surrounding central nuclei or surrounding nuclei located at the surface of regenerated fibers. No labeling of perinuclear regions of control muscle or prenecrotic fibers was observed. Myonuclei fractionated from adult mdx hindlimb muscles contained no PDGF-receptor, indicating that PDGF-receptor-positive structures are not tightly associated with nuclei or within nuclei. L6 myoblasts show PDGF-receptor distributed diffusely on the cell surface. Stimulation of L6 myoblasts with 10 ng/ml of PDGF-BB causes receptor internalization and concentration in granules at perinuclear regions. Thus, PDGF stimulation of myoblasts causes a redistribution of PDGF-receptors to resemble receptor localization observed during muscle regeneration. These findings implicate PDGF-mediated mechanisms in regeneration of dystrophic muscle.

Amino Acid Sequence↗