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Morphological and physiological properties of non-striated muscle from the tunicate, Ciona intestinalis: parallels with vertebrate skeletal muscle.

Non-striated muscle from the longitudinal body wall muscle in an adult ascidian (tunicate) is characterized morphologically and physiologically. These muscles are unique among chordate non-striated (i.e. 'smooth') muscles in that they are composed of discrete bundles of several small diameter (4-10 microns) muscle cells (fibers) arranged in parallel functional units. Each bundle is wrapped by a basal lamina, and at least some appear to be directly innervated at a neuromuscular junction similar to an end plate. In these regards, a bundle of Ciona smooth muscle cells is analogous to a skeletal muscle fiber of a vertebrate. This analogy also extends to physiological properties. Ciona muscle generates a rapid all-or-none Ca action potential which gives rise to a brisk twitch with brief latency. These anatomical and physiological adaptations are discussed in terms of the evolution of vertebrate skeletal muscle.

Action Potentials↗

In vitro transdifferentiation of striated muscle to smooth muscle cells of a medusa.

Mononucleated striated muscle cells can be isolated from anthomedusae and cultivated in artificial seawater. In the cultivated muscle the differentiated state is maintained and DNA synthesis is not observed. The isolated striated muscle can be activated by collagenase treatment to transdifferentiate into various new cell types. Between the second and third day following collagenase treatment DNA synthesis is initiated, and mitosis and de novo flagellum formation occur in the isolated muscle. Under these circumstances all isolated striated muscle fragments produce both smooth muscle cells and y-cells (Schmid and Alder, 1984). In experiments, in which either transcription (actinomycin D) or translation (cycloheximide) is inhibited, the activated striated muscle cells do not transdifferentiate but maintain their differentiated state. Inhibition of DNA replication (aphidicolin), however, results in uniform transdifferentiation of striated muscle to smooth muscle cells in the absence of y-cell types (Schmid and Alder, 1984). The fluorescence stain NBD-phallacidin is used to monitor the characteristic change of F-actin pattern of these isolates.

Actins↗

Estimation of average muscle fiber conduction velocity from simulated surface EMG in pinnate muscles.

The aim of this simulation study was to assess the bias in estimating muscle fiber conduction velocity (CV) from surface electromyographic (EMG) signals in muscles with one and two pinnation angles. The volume conductor was a layered medium simulating anisotropic muscle tissue and isotropic homogeneous subcutaneous tissue. The muscle tissue was homogeneous for one pinnation angle and inhomogeneous for bipinnate muscles (two fiber directions). Interference EMG signals were obtained by simulating recruitment thresholds and discharge patterns of a set of 100 and 200 motor units for the pinnate and bipinnate muscle, respectively (15 degrees pinnation angel in both cases). Without subcutaneous layer and muscle fibers with CV 4m/s, average CV estimates from the pinnate (bipinnate) muscle were 4.81+/-0.18 m/s (4.80+/-0.18 m/s) for bipolar, 4.71+/-0.19 m/s (4.71+/-0.12 m/s) for double differential, and 4.78+/-0.16 m/s (4.79+/-0.15m/s) for Laplacian recordings. When subcutaneous layer was added (thickness 1mm) in the same conditions, estimated CV values were 4.93+/-0.25 m/s (5.16+/-0.41 m/s), 4.70+/-0.21 m/s (4.83+/-0.33 m/s), and 4.89+/-0.21 m/s (4.99+/-0.39 m/s), for the three recording systems, respectively. The main factor biasing CV estimates was the propagation of action potentials in the two directions which influenced the recording due to the scatter of the projection of end-plate and tendon locations along the fiber direction, as a consequence of pinnation. The same problem arises in muscles with the line of innervation zone locations not perpendicular to fiber direction. These results indicate an important limitation in reliability of CV estimates from the interference EMG when the innervation zone and tendon locations are not distributed perpendicular to fiber direction.

Action Potentials↗

Immunohistochemical localization of alpha(1a)-adrenoreceptors in muscle spindles of rabbit masseter muscle.

The expression of alpha(1a)-adrenoreceptors (alpha(1a)-ARs) within the muscle spindles of rabbit masseter muscle was investigated. The alpha(1a)-ARs were detected by immunohistochemical fluorescent method and examined along the entire length of 109 cross serially sectioned spindles. The sympathetic fibers were visualized by the immunofluorescent labeling of the noradrenaline synthesizing enzymes tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). In order to recognize the intrafusal muscle fiber types, antibodies for different myosin heavy chain isoforms (MyHCI) were used. TH and DBH immunolabeled nerve fibers have been observed within the capsule lamellar layers, in the periaxial fluid space and close to intrafusal muscle fibers. The alpha(1a)-ARs were detected on the smooth muscle cells of the blood vessels coursing in the muscle and in the capsule lamellar layers or within the periaxial fluid space of the spindles. Moreover, at the polar regions of a high percentage (88.1%) of muscle spindles a strong alpha(1a)-ARs immunoreactivity was present on the intrafusal muscle fibers. In double immunostained sections for alpha(1a)-ARs and MyHCI it was evidenced that both bag, and nuclear chain fibers express alpha(1a)-ARs. The receptors that we have detected by immunofluorescence may support a direct control by adrenergic fibers on muscle spindle.

Animals↗

Biological progression from adult bone marrow to mononucleate muscle stem cell to multinucleate muscle fiber in response to injury.

Adult bone marrow-derived cells (BMDC) are shown to contribute to muscle tissue in a step-wise biological progression. Following irradiation-induced damage, transplanted GFP-labeled BMDC become satellite cells: membrane-ensheathed mononucleate muscle stem cells. Following a subsequent exercise-induced damage, GFP-labeled multinucleate myofibers are detected. Isolated GFP-labeled satellite cells are heritably myogenic. They express three characteristic muscle markers, are karyotypically diploid, and form clones that can fuse into multinucleate cells in culture or into myofibers after injection into mouse muscles. These results suggest that two temporally distinct injury-related signals first induce BMDC to occupy the muscle stem cell niche and then to help regenerate mature muscle fibers. The stress-induced progression of BMDC to muscle satellite cell to muscle fiber results in a contribution to as many as 3.5% of muscle fibers and is due to developmental plasticity in response to environmental cues.

Animals↗

Differential innervation of the goldfish tonic red muscles and twitch white muscles by neuropeptide-immunoreactive motoneurons.

Neuropeptides in the motor nerves innervating the red and white muscles of the goldfish Carassius auratus were examined. In the tonic red muscles, varicose nerve endings immunoreactive for both calcitonin gene-related peptide and substance P were found spread over the surface of the muscle fibers, but in the twitch white muscles only scattered nerve endings immunoreactive for calcitonin gene-related peptide were found. At the electron microscopic observation, dense electron products immunoreactive for calcitonin gene-related peptide and for substance P (SP) were detected in the motor nerve endings making synapses on the muscle fibers of the red muscles. In the spinal cord, all of the motor neurons showed immunoreactivity to calcitonin gene-related peptide, but the motor neurons immunoreactive for substance P were restricted to the ventrolateral group that has been shown to project predominantly to the red muscles. These results suggest that the motor neurons innervating the red and white muscles of the goldfish are distinct in their neuropeptide content. The present study also raises the possibility that SP might be related to the unique physiological properties of the tonic type red muscles, probably by direct binding to the acetylcholine receptors.

Animals↗

Muscle activity-resistant acetylcholine receptor accumulation is induced in places of former motor endplates in ectopically innervated regenerating rat muscles.

Expression of acetylcholine receptors (AChRs) in the extrajunctional muscle regions, but not in the neuromuscular junctions, is repressed by propagated electric activity in muscle fibers. During regeneration, subsynaptic-like specializations accumulating AChRs are induced in new myotubes by agrin attached to the synaptic basal lamina at the places of former motor endplates even in the absence of innervation. We examined whether AChRs still accumulated at these places when the regenerating muscles were ectopically innervated and the former synaptic places became extrajunctional. Rat soleus muscles were injured by bupivacaine and ischemia to produce complete myofiber degeneration. The soleus muscle nerve was permanently severed and the muscle was ectopically innervated by the peroneal nerve a few millimeters away from the former junctional region. After 4 weeks of regeneration, the muscles contracted upon nerve stimulation, showed little atrophy and the cross-section areas of their fibers were completely above the range in non-innervated regenerating muscles, indicating successful innervation. Subsynaptic-like specializations in the former junctional region still accumulated AChRs (and acetylcholinesterase) although no motor nerve endings were observed in their vicinity and the cross-section area of their fibers clearly demonstrated that they were ectopically innervated. We conclude that the expression of AChRs at the places of the former neuromuscular junctions in the ectopically innervated regenerated soleus muscles is activity-independent.

Acetylcholinesterase↗

Muscle development, insulin-like growth factor-I and myostatin mRNA levels in chickens selected for increased breast muscle yield.

Insulin-like growth factor-I (IGF-I) and myostatin (MSTN) are paracrine regulators of muscle growth. The present study was conducted to relate their expression with muscle fibre development in chickens selected for high breast meat yield and their controls. Both mRNA levels were measured by real-time RT-PCR in the Pectoralis major (PM) muscle between 14 days in ovo and 6 weeks post-hatch and in the Sartorius (SART) muscle between 2 and 6 weeks. The data show that PM growth was slow during in ovo development and rapid in the early post-hatch period. Chickens from the selected genotype exhibited significantly higher breast muscle yields from 2 to 6 weeks of age, and muscle fibre hypertrophy. In the PM, IGF-I and MSTN mRNA levels decreased markedly around hatch, while the IGF-I/MSTN ratio increased, suggesting that it could contribute to the explosive growth observed in the early post-hatch period. Between 4 and 6 weeks of age in selected chickens, IGF-I mRNA levels were significantly higher (p=0.04) with a similar trend in MSTN mRNA levels (p=0.07) in the PM muscle but not in the SART muscle. Our results support the hypothesis that the relative levels of IGF-I and MSTN mRNA may participate to set muscle growth rate along development, while other factors are required to explain differences between genotypes.

Animals↗

Quadriceps concentric and eccentric exercise 2: differences in muscle strength, fatigue and EMG activity in eccentrically-exercised sore and non-sore muscles.

The purpose of this study was to determine whether the previously reported muscle weakness and increases in EMG and EMG/force ratio after eccentric exercise were related to ensuing soreness or simply to the exercise mode. Delayed-onset muscle soreness, maximal voluntary force and the surface electromyogram (EMG) of the quadriceps were studied in 10 healthy male subjects following 20 min of bench-stepping with a constant leading leg. Prior to stepping and at 0, 0.25, 0.50, 0.75, 1, 3, 24 and 48 h afterwards the subjects performed a 30 s isometric maximal voluntary contraction (MVC) of the extensors of each leg during which the isometric force and the root mean square voltage of the surface EMG (rms EMG) were recorded. Muscle soreness was recorded prior to and at 0, 0.75, 3, 24 and 48 h after the stepping task by means of a force probe. No soreness was experienced in the concentrically exercised muscles, but in the eccentrically exercised muscles 5 subjects were not sore and 5 experienced soreness after 24 h, increasing to 48 h. There were no significant differences between sore and non-sore muscles in muscle weakness, fatigue during 30 s, the EMG amplitudes or the EMG/force ratios during peak force and the 30 s MVC (p >0.05). Muscle weakness following eccentric exercise was related to the exercise mode and was independent of subsequent soreness. Both weakness and soreness may be related to muscle damage, but involve different mechanisms.

Adult↗

Immunocytochemical electron microscopic study and western blot analysis of myosin, paramyosin and miniparamyosin in the striated muscle of the fruit fly Drosophila melanogaster and in obliquely striated and smooth muscles of the earthworm Eisenia foetida.

Miniparamyosin is a paramyosin isoform (55-60 kDa) that has been isolated in insects (Drosophila) and immunolocalized in several species of arthropods, molluscs, annelids and nematodes. In this study, the presence and distribution of this protein, in comparison with that of paramyosin and myosin, has been examined in the striated muscle (tergal depressor of trochanter) of Drosophila melanogaster, and the obliquely striated muscle (body wall) and the smooth muscle (outer layer of the pseudoheart) of the earthworm Eisenia foetida by means of immunocytochemical electron microscopic study and Western blot analysis miniparamyosin paramyosin and myosin antibodies from Drosophila. In the striated muscle of D. melanogaster, the three proteins were immunolocalized along the length of the thick filaments (A-bands). The distribution of immunogold particles along these filaments was uniform. The relative proportions miniparamyosin/paramyosin/myosin (calculated by counting the number of immunogold particles) were: 1/10/68. In the obliquely striated muscle of E. foetida, immunoreactions to the three proteins were also found in the thick filaments, and the relative proportions miniparamyosin/paramyosin/myosin were 1/2.4/6.9. However, whereas the distribution of both myosin and miniparamyosin along the thick filament length was uniform, paramyosin immunolabelling was more abundant in the extremes of thick filaments (the outer zones of A-bands in the obliquely striated muscle), where the thick filaments become thinner than in the centre (the central zone of A-bands), where these filaments are thicker. The relative proportions of paramyosin in the outer and of paramyosin in the central zones of A-bands were 4/1. This irregular distribution of paramyosin along the thick filament length might be actual but it may also be explained by the fusiform shape of thick filaments in the earthworm: assuming that paramyosin is covered by myosin, paramyosin antigens would be more exposed in the tips than in the centre of thick filaments. If miniparamyosin is, in turn, covered by paramyosin, the exposure of miniparamyosin antigens would be low even in the tips of thick filaments, and this might explain the scanty immunoreaction observed for this protein and the absence of a higher number of immunogold particles in the extremes of thick filaments. The distribution of the three proteins in the earthworm smooth muscle was as in the obliquely striated muscle, although the proportions miniparamyosin/paramyosin/myosin were 1/1.5/5.2; this is, immunoreactions to paramyosin and miniparamyosin were lower than in the obliquely striated muscle.

Animals↗

The activation of protein degradation in muscle by Ca2+ or muscle injury does not involve a lysosomal mechanism.

By use of different inhibitors, we distinguished three proteolytic processes in rat skeletal muscle. When soleus muscles maintained under tension were exposed to the calcium ionophore A23187 or were incubated under no tension in the presence of Ca2+, net protein breakdown increased by 50-80%. Although leupeptin and E-64 inhibit this acceleration of protein breakdown almost completely, other agents that prevent lysosomal function, such as methylamine or leucine methyl ester, did not inhibit this effect. A similar increase in net proteolysis occurred in muscle fibres injured by cutting, and this response was also inhibited by leupeptin, but not by methylamine. In contrast, all these inhibitors markedly decreased the 2-fold increase in protein breakdown induced by incubating muscles without insulin and leucine, isoleucine and valine. In addition, the low rate of proteolysis seen in muscles under passive tension in complete medium was not affected by any of these inhibitors. Thus the basal degradative process in muscle does not involve lysosomes or thiol proteinases, and muscle can enhance protein breakdown by two mechanisms: lack of insulin and nutrients enhances a lysosomal process in muscle, as in other cells, whereas Ca2+ and muscle injury activate a distinct pathway involving cytosolic thiol proteinase(s).

Animals↗

Limits of muscle-to-nerve ratio in functional muscle transplantation.

The aim of the present study was to investigate the functional limits of over-dimensioning a free functioning muscle transplant for neuromuscular reconstruction. A part of the rectus femoris muscle, two and three times bigger than a scutuloauricularis muscle, was used to functionally replace the latter after removal. The major finding of these experiments is that both double-sized and triple-sized portions of the rectus femoris muscle developed maximal tetanic tensions during isometric contractions, which were up to 175 percent of the control scutuloauricularis muscle of the unoperated, contralateral side, although the same branch of the facial nerve was used for reinnervation of the grafted muscle. This implies that the supplying branch of the facial nerve has the potency to innervate a muscle much larger than the originally supplied muscle with optimal efficiency. These results underline the usefulness of overdimensioning during functional muscle transplantation, and also in limited neural capacity situations.

Animals↗

Growth and metabolism of fetal and maternal muscles of adolescent sheep on adequate or high feed intake: possible role of protein kinase C-alpha in fetal muscle growth.

From days 4-104 of pregnancy, adolescent sheep, weighing 43.7 (SE 0.87) kg were offered a complete diet at two different intakes (approximately 5 or 15 kg/week) designed to meet slightly, or well above, maternal maintenance requirements. The fetal and maternal muscles were taken on day 104 of pregnancy and analysed for total DNA, RNA and protein. Ewes offered a high intake to promote rapid maternal weight gain, weighed more (76.5 (SE 4.5) v 50.0 (SE 1.7) kg) and had muscles with a greater fresh weight, whilst their fetuses had smaller muscles, than those fed at a lower intake. Plantaris muscle of the ewes fed at the high intake contained more RNA and protein; again the opposite situation was found in the fetal muscle. On the higher maternal intakes, the DNA, RNA and protein contents of the fetal plantaris muscle were less than in fetuses of ewes fed at the lower intake. To investigate the possible mechanisms involved in this decrease in fetal muscle mass, cytosolic and membrane-associated muscle proteins were subjected to Western immunoblotting with antibodies to nine isoforms of protein kinase C (PKC), a family of enzymes known to play an important role in cell growth. Five PKC isoforms (alpha, epsilon, theta, mu, zeta) were identified in fetal muscle. One of these, PKC-alpha was located predominantly in the cytosolic compartment in the smaller fetuses of the ewes fed at a high plane of nutrition, but was present to a greater extent in the membranes of the more rapidly growing fetuses of the ewes fed at the lower intake. This was the only isoform to demonstrate nutritionally related changes in it subcellular compartmentation suggesting that it may mediate some aspects of the change in fetal growth rate.

Animal Nutritional Physiological Phenomena↗

The influence of undernutrition during gestation on skeletal muscle cellularity and on the expression of genes that control muscle growth.

We examined the effects of two levels of gestational undernutrition (50 % and 40 % of ad libitum) on postnatal growth rate, skeletal muscle cellularity and the expression of genes that control muscle growth, in the offspring at weaning. The results showed that the rat pups born to mothers fed the 50 % diet during gestation and a control diet during lactation had an increased postnatal growth rate compared with the pups fed the more restricted diet (40 % of ad libitum). Surprisingly, the growth rate of the control group (ad libitum) was intermediate between the 50 % and 40 % groups. The restricted diets did not alter the number of muscle fibres in the semitendinosus muscle of the offspring but the number of muscle nuclei was reduced by 16 % in the 40 % group compared with the control group. In the 50 % group, the lightest pups at birth (L) had elevated muscle insulin-like growth factor (IGF)-1, IGF binding protein (BP)-5 and proliferating cell nuclear antigen (PCNA) mRNA compared with the L pups from both the control and 40 % groups. The heaviest pups at birth (H) in the 50 % group had increased levels of IGFBP-4, PCNA and M-cadherin mRNA compared with both the control and 40 % groups. Levels of IGF-1 receptor, myostatin and MyoD mRNA did not correlate with postnatal growth. Both H and L pups from the 40 % group had reduced muscle IGF-1 mRNA but all other transcripts examined were similar to control levels. The results suggest that the increased postnatal growth rate, which accompanied milder fetal undernutrition (50 %), may be due to a more active local muscle IGF system and increased muscle-cell proliferation.

Animals↗

Effects of aging on muscle T2 relaxation time: difference between fast- and slow-twitch muscles.

RATIONALE AND OBJECTIVES: To determine whether the T2 relaxation time of skeletal muscle is affected by aging and to compare the effects of aging between fast- and slow-twitch muscles in a human study. To investigate the mechanisms of age-related changes in T2 relaxation time in an animal (mouse) study. METHODS: T2 relaxation times of the soleus (slow-twitch, rich in type I fiber) and gastrocnemius (fast-twitch, rich in type II fiber) muscles were examined in 59 healthy human subjects, 22 to 76 years of age, by clinical magnetic resonance imaging. In mice, T2 relaxation times, fat ratios, and extracellular space ratios (extracellular space/intracellular plus extracellular space) of the spinalis (fast-twitch, rich in type II fiber) muscles were also examined (group of 7 old mice, 24-26 months; group of 7 young mice, 8-10 weeks). RESULTS: In the human study, the T2 relaxation time of the gastrocnemius muscle increased significantly with aging (r = 0.53, P < 0.01) while that of the soleus muscle did not. In the animal study, the T2 relaxation time of the spinalis muscle was significantly longer (P < 0.05) and the extracellular space ratio of the spinalis muscle significantly wider (P < 0.01) in old than in young mice. No significant difference in fat ratio was observed between old and young mice. A significant, positive correlation was seen between the extracellular space ratio and T2 relaxation time (r = 0.84, P < 0.01). CONCLUSIONS: The T2 relaxation time of fast-twitch muscle increases with aging, due mainly to increased extracellular space, reflecting age-related type II fiber atrophy.

Adult↗

Experimental studies on masseter muscle grafts in rats: comparison of donor muscles.

Experimental studies were performed in rats to explore the feasibility of using limb muscles to replace masticatory masseter muscles. The results showed that if several small fusiform muscles such as the extensor digitorum longus (EDL), soleus (SOL) and plantaris (PLT) were used for the grafting, the graft regenerated and matured fully in the masseter bed, although the weights were lower than those of contralateral control masseter muscles. If a multipennate muscle such as the gastrocnemius was used for the grafting, the graft also regenerated but later degenerated due to the lack of innervation. This is probably due to the excessive amount of connective tissue in the multipennate muscle. The connective tissue proliferates faster than the regeneration of muscle fibres and, thus, renders the innervation of muscle fibres difficult. Other factors pertinent to the possible clinical application of muscle transplantation in oral region are discussed.

Animals↗

Changes in pennation with joint angle and muscle torque: in vivo measurements in human brachialis muscle.

1. Estimates of pennation in human muscles are usually obtained from cadavers. In this study, pennation of human brachialis was measured in vivo using sonography. Effects of static and dynamic changes in elbow angle and torque were investigated. 2. Pennation was measured in eight subjects using an 80 mm, 5 MHz, linear-array ultrasound transducer to generate sagittal images of the brachialis during maximal and submaximal isometric contractions at various elbow angles. It was shown that estimates of pennation were reproducible, representative of measurements made throughout the belly of the muscle and not distorted by compression of the muscle with the transducer or rotation of the muscle out of the plane of the transducer. 3. Mean resting pennation was 9.0 +/- 2.0 deg (S.D., range 6.5-12.9 deg). When the muscle was relaxed there was no effect of elbow angle on pennation. However, during a maximal isometric contraction (MVC), with the elbow flexed to 90 deg, pennation increased non-linearly with elbow torque to between 22 and 30 deg (mean 24.7 +/- 2.4 deg). The effect of increasing torque was small when the elbow was fully extended. The relationship between elbow angle, elbow torque and brachialis pennation suggests that the relaxed brachialis muscle is slack over much of its physiological range of lengths. 4. There was no hysteresis in the relationship between torque and pennation during slow isometric contractions (0.2 MVC s-1), and the relationship between elbow angle and pennation was similar during slow shortening and lengthening contractions. 5. Two consequences follow from these findings. Firstly, intramuscular mechanics are complex and simple planar models of muscles underestimate the increases in pennation which occur during muscle contraction. Second, spindle afferents from relaxed muscles may not encode joint angle over the full range of movement.

Adult↗

Smooth muscle myosin light chain kinase expression in cardiac and skeletal muscle.

The purpose of this study was to characterize myosin light chain kinase (MLCK) expression in cardiac and skeletal muscle. The only classic MLCK detected in cardiac tissue, purified cardiac myocytes, and in a cardiac myocyte cell line (AT1) was identical to the 130-kDa smooth muscle MLCK (smMLCK). A complex pattern of MLCK expression was observed during differentiation of skeletal muscle in which the 220-kDa-long or "nonmuscle" form of MLCK is expressed in undifferentiated myoblasts. Subsequently, during myoblast differentiation, expression of the 220-kDa MLCK declines and expression of this form is replaced by the 130-kDa smMLCK and a skeletal muscle-specific isoform, skMLCK in adult skeletal muscle. These results demonstrate that the skMLCK is the only tissue-specific MLCK, being expressed in adult skeletal muscle but not in cardiac, smooth, or nonmuscle tissues. In contrast, the 130-kDa smMLCK is ubiquitous in all adult tissues, including skeletal and cardiac muscle, demonstrating that, although the 130-kDa smMLCK is expressed at highest levels in smooth muscle tissues, it is not a smooth muscle-specific protein.

Amino Acid Sequence↗