Histochemical studies on the amphibian opercularis muscle (Amphibia)

Summary Histochemical studies of the opercularis muscle of the bullfrog (Rana catesbeiana) and the tiger salamander (Ambystoma tigrinum) provide evidence that the opercularis muscle of anurans is a specialized, tonic portion of the levator scapulae superior muscle. Staining results for myosin adenosine triphosphatase (ATPase) and succinate dehydrogenase (SDH), combined with measurements of muscle fiber diameters, demonstrate that the opercularis/levator scapulae superior muscle mass of both the tiger salamander and bullfrog consists of an anterior tonic portion, a middle fast oxidative-glycolytic (FOG) twitch portion, and a posterior fast-glycolytic (FG) twitch portion. In R. catesbeiana the tonic fibers represent 57.3% of the fiber total and (because they have relatively narrow diameters) about 29% of the cross-sectional area of the muscle mass, and form that part of the muscle (=opercularis muscle) that inserts on the operculum. In Ambystoma the tonic fibers represent only 8.8% of the fiber total and represent about 4% of the cross-sectional area. In the tiger salamander, the entire levator scapulae superior muscle inserts on the operculum and therefore represents the opercularis muscle. The bullfrog differs from the tiger salamander, therefore, in that the anterior tonic part of the opercularis/levator scapulae superior complex is greatly enlarged and the insertion on the operculum is limited to these tonic fibers. No evidence of a columellar muscle was found in R. catesbeiana. Previous reports of one in this species and in other anurans may be based on the tripartite nature of the opercularis/levator scapulae superior muscle mass. The middle FOG portion of the muscle may have been considered a muscle distinct from the anterior tonic portion (=opercularis muscle) and the posterior FG portion.     

Role of the opercularis muscle in seismic sensitivity in the bullfrog Rana catesbeiana

The inner ear of anuran amphibians appears to be exceptionally sensitive to substrate vibration. The opercularis system, consisting of an opercularis muscle running from the shoulder girdle to a movable, cartilaginous operculum lying next to the inner ear, has been hypothesized to be involved in driving these seismic responses. Removal of the opercularis muscle of adult bullfrogs, Rana catesbeiana, caused clear decreases in microphonic responses of the inner ear to vibrations from 20-250 Hz and 0.05-5.0 cm/sec2 accelerations. Degree of decrease in responsiveness was variable between individuals and between different frequencies of stimulation, ranging up to 90% reduction at certain frequencies and in certain specimens. Decreases were most marked at lower frequencies below about 50 Hz. Additional removal of the levator scapulae superior muscle, which runs alongside the opercularis muscle from the shoulder girdle to ventrolateral portions of the otic capsule, also tended to depress responses, although this effect was substantially less (generally less than 10%) and also less consistent. As the opercularis muscle appears to be derived from the levator scapulae musculature, it is speculated that primitively seismic sensitivity was enhanced by a muscular connection that could transmit motion from the forelimb to the otic region, responsiveness being further enhanced by the subsequent evolution of the specialized opercularis system.     

Properties of standard avian slow muscle grafts following long-term regeneration

The tonic anterior latissimus dorsi (ALD) of adult pigeons was orthotopically homografted and evaluated after 11 months of regeneration for histological, histochemical, electromyographic (EMG), and mechanical properties. The resting EMG activity of the grafts was lower in amplitude than that of the controls, but showed the tonic pattern typical for these tonic muscles. The control and grafted muscles had a histochemically homogeneous population of fibers with moderate myofibrillar adenosine triphosphatase activity. Succinic dehydrogenase activity was moderate for the control muscles, but low for the grafts. The regenerated muscles had fewer and smaller fibers and had much larger intersynaptic distances. Both the regenerated and the contralateral control muscles were slow contracting and maintained tetanic tension for prolonged periods with direct electrical stimulation. The relaxation was slower in the grafted muscle than in the control. The grafts produced 40% of the maximum tension of the control muscles, but the rate of tension development was similar between the two groups. The results indicate that the tonic properties were regenerated, but the innervation pattern was altered and the grafted muscles did not have normal mature fibers even after long-term regeneration.     

Timing of development of the middle ear of Anura (Amphibia)

Summary The opercularis system and tympanum-stapes complex of the anuran middle ear develop at different times relative to metamorphosis. In early larvae, the fenestra ovalis is represented by a large lateral opening in the otic capsule filled with connective tissue. At later larval stages, but well before metamorphosis, a cartilaginous operculum begins to form at the posterior margin of the fenestra ovalis, and proceeds to expand to fill all except the anterior part of the fenestra. The opercularis muscle forms along with the levator scapulae superior muscle at the anteromedial edge of the developing suprascapular cartilage of the shoulder girdle. The muscle fibers extend anteroventrally towards the operculum and otic capsule, and, just before emergence of the forelimbs, that portion that will form the opercularis muscle inserts on the lateral surface of the operculum. At this stage, when the metamorphosing frogs first show terrestrial habits, the opercularis system is complete and presumably functional. Timing of development of the tympanum-stapes complex is more variable. The stapes begins as a cartilaginous condensation in the anterior part of the fenestra ovalis, and develops laterally to eventually contact the epidermis and dermis that together will form the tympanum. Meanwhile a middle ear cavity and tympanic annulus form to complete the complex. In several species, especially those that metamorphose at a smaller body size, the tympanum-stapes complex is quite incomplete by the end of metamorphosis, and in Hyla crucifer it takes about 60 days to fully develop. The presence of a complete opercularis system by the start of terrestrial activity is consistent with an hypothesized seismic function of the system. The independent timing of development of the opercularis system and tympanum-stapes complex does not support functional hypotheses linking the opercularis system with modulation of responsiveness of the tympanum-stapes complex to aerial sound. Newly metamorphosed frogs with poorly developed tympanum-stapes complexes are presumably either insensitive to aerial sound or employ alternate mechanisms for transmission of sound energy to the inner ear, possibly involving the opercularis system.     

Distribution and morphology of motoneurons innervating different muscle fiber types in an amphibian muscle complex

The distribution and morphology of motoneurons innervating specific types of muscle fibers in the levator scapulae superior (LSS) muscle complex of the bullfrog (Rana catesbeiana) and tiger salamander (Ambystoma tigrinum) were studied by retrograde labelling with cholera toxin-conjugated horseradish peroxidase (CT-HRP). The LSS muscle complex in both of these amphibians has a segregated pattern of muscle-fiber types (tonic; fast oxidative-glycolytic twitch [FOG]; fast glycolytic twitch [FG]) along an anteroposterior axis. The entire motor pool was labelled by injection of CT-HRP into the whole LSS muscle complex. The motoneurons innervating specific fiber types were labelled by injection of CT-HRP into certain muscle regions. The organization of the motoneuron pool of the LSS complex of both species was arranged in two columns—one ventrolateral and one medial. In bullfrogs, the ventrolateral column contains motoneurons innervating FG and tonic fiber types and the medial column contains motoneurons innervating FOG fiber types. In tiger salamanders, the ventrolateral column contains motoneurons innervating FG fiber types and the medial column contains motoneurons innervating FOG and tonic fiber types. The different motoneuron types also have different soma sizes and patterns of dendritic arborization. In both species, FG motoneurons are the largest, whereas FOG motoneurons are intermediate in size and tonic motoneurons are the smallest. In bullfrogs, the main dendrites of FG motoneurons extend into the dorsolateral and the ventrolateral gray matter of the spinal cord, whereas the dendrites of FOG motoneurons extend into the ventral and medial cord. In the tiger salamander, dendrites of FG motoneurons extend into the ventrolateral spinal cord and dendrites of the FOG motoneurons extend more generally into the ventral cord. Dendrites of tonic motoneurons in both amphibians were small and short, and difficult to observe. These results establish that motoneurons innervating different types of muscle fibers in the LSS muscle complex are segregated spatially and display consistent morphological differences. © 1993 Wiley-Liss, Inc.     

Low frequency sounds from sustained contraction of human skeletal muscle.

Low frequency audible vibrations are produced by human skeletal muscles undergoing sustained contraction. The effect is easily demonstrable with an electronic stethoscope which amplifies sound below 50 Hz. Autocorrelation analysis of the signal shows that it is periodic with a frequency 25 +/- 2.5 Hz. The quality of the sound is the same for all the skeletal muscles tested and is unaffected by changes in tension, ambient temperature, and blood flow. Electrically-stimulated contraction produces a sound which is indistinguishable from voluntary contraction. The amplitude of the sound increases linearly with tension. The sound signals are uncorrelated both in frequency and phase with electromyographic signals obtained simultaneously while the muscle is contacted. Arguments are presented to show that the sounds may be an intrinsic property of muscle contraction.     

Function of Peripheral Inhibitory Axons in Insects

There are now many examples in insects of axons which elicithyperpolarizing junctional potentials in the muscle fibers theyinnervate. With the muscles bathed in haemolymph, electricalstimulation of these axons causes a decrease in the magnitudeof slow contractions. This property allows them to be definedas inhibitory. Although inhibitory axons have the ability toregulate the magnitude of maintained slow contractions, thereis little evidence that this is their normal function. The inhibitoryaxons supplying at least three insect muscles function to increasethe rate of relaxations following each contraction of a rhythmicsequence. Moreover, when the haemolymph potassium concentrationis high, some inhibitory axons probably ensure complete relaxationbetween rhythmic contractions by preventing potassium contractionsin tonic muscle fibers. There is no convincing evidence thatinhibitory axons can facilitate muscular contractions by becomingactive immediately before the excitatory input.     

Pre-imaginal flight motor pattern in Locusta

In a wind stream, larval stages of Locusta usually show a tonic muscle activity but they can also exhibit a rhythmic motor output. With ageing such a pattern can be released sooner, the trains become longer. The basic rhythm of 10 Hz does not change. The initial co-contraction of specific muscles is substituted later in development by an antagonistic recruitment. This activity resembles the flight motor pattern of young locusts which lack phasic sensory feedback from the wing region. Azadirachtin, an insect growth regulator, has been used to produce a permanent 5th larval instar. However, the extension of the last larval stage does not lead to a further development of the motor pattern to a level comparable to mature animals.     

The use of opercular muscle electromyograms as an indicator of the metabolic costs of fish activity in rainbow trout, Salmo gairdneri Richardson, as determined by radiotelemetry

Averages of electromyogram (EMG) signals emanating from the levator arcus palatini , a small muscle involved in the operation of the operculum in rainbow trout, Salmo gairdneri , were analysed in terms of their relationship to the fish's oxygen consumption rates under various activity levels. The EMG signals were detected and transmitted with a radio-telemetry system. The EMG values showed a good correlation with corresponding oxygen consumption rates for fish under forced-swimming conditions but not when the fish was swimming spontaneously; this is attributed to an ability to regulate oxygen uptake at the gill surfaces by other means than increasing the ventilation volume, including alterations in the gill blood flow dynamics (e.g. secondary lamellar recruitment), and changes in the cardiac output. Under forced-swim conditions, where the oxygen demands by the respiring muscles were higher, increased ventilation volume, as indicated by increased opercular muscle activity, was directly related to swimming speed and oxygen uptake.     

The effect of postural correction on muscle activation amplitudes recorded from the cervicobrachial region

In clinical practice, postural correction is a common treatment approach for individuals with neck and shoulder pain. As chronic static muscle use is thought to be associated with the onset of some neck and shoulder pain syndromes, it is important to understand the impact a postural correction program might have on muscle activation amplitudes in the neck and shoulder regions. Normalized surface electromyographic data were recorded from the levator scapulae, upper trapezius, supraspinatus, posterior deltoid, masseter, rhomboid major, cervical erector spinae, and sternocleidomastoid muscles of the dominant side of each of eighteen healthy subjects. Subjects performed five repetitions of each of four seated typing postures (habitual, corrected, head-forward and slouched) and four standing postures (habitual, corrected, and head-forward and slouched). Repeated-measures analysis of variance models (α = 0.05) revealed that in sitting postural correction tended to decreased the level of muscle activation required in all muscles studied during seated computer work, however this finding was not statistically significant. Corrected posture in sitting did, however produce a statistically significant reduction in muscle activity compared to forward head posture. Corrected posture in standing required more muscle activity than habitual or forward head posture in the majority of cervicobrachial and jaw muscles, suggesting that a graduated approach to postural correction exercises might be required in order to train the muscles to appropriately withstand the requirements of the task. A surprising finding was that muscle activity levels and postural changes had the largest impact on the masseter muscle, which demonstrated activation levels in the order of 20% maximum voluntary electrical activation.     

Morphofunctional and histochemical characteristics of the hindlimb muscles of the Rana temporaria in ontogeny

Mixed muscles of adult frogs respond to the increase in external potassium and to Ach by polyphasic contracture which is due to asynchronous activity of various groups of muscle fibers (fast phasic, intermediate and tonic ones). In the developing in vivo hindlimb muscles, the predominance of phasic contractile response and relatively weak tonic one were noted. In contrast to definitive muscles, in which maximum potassium and acetylcholine contractures are identical, growing muscles produce weak contractile reaction to Ach. Ach sensitivity of the developing muscles (as revealed by the contracture) is lower than in the definitive ones. Histochemical (studies on the lipid content and the activity of succinate dehydrogenase) and morphometric (the ratio of muscle fibers of different types at different stages of development, comparison of their diameters, relative size of tonic bundle, etc.) studies indicate that the development of morphological substrate for tonic contractions (tonic and intermediate muscle fibers) takes place at a lower rate as compared to the development of the substrate for phasic contractions. However, histochemically tonic fibers may be revealed already at the stage of myotubes.     

Segment-specific retention of a larval neuromuscular system and its role in a new, rhythmic, pupal motor pattern in Manduca sexta

At pupation in Manduca sexta, accessory planta retractor muscles and their motoneurons degenerate in segment-specific patterns. Accessory planta retractor muscles in abdominal segments 2 and 3 survive in reduced form through the pupal stage and degenerate after adult emergence. Electromyographic and electrophysiological recordings show that these accessory planta retractor muscles participate in a new, rhythmic `pupal motor pattern' in which all four muscles contract synchronously at ∼4 s intervals for extended bouts. Accessory planta retractor muscle contractions are driven by synaptic activation of accessory planta retractor motoneurons and are often accompanied by rhythmic activity in intersegmental muscles and spiracular closer muscles. The pupal motor pattern is influenced by descending neural input although isolated abdominal ganglia can produce a pupal motor pattern-like rhythm. The robust pupal motor pattern first seen after pupal ecdysis weakens during the second half of pupal life. Anemometric recordings indicate that the intersegmental muscle and spiracular closer muscle component of the pupal motor pattern produces ventilation. Accessory planta retractor muscle contractions lift the flexible abdominal floor, to which the developing wings and legs adhere tightly. We hypothesize that, by a bellows-like action, the accessory planta retractor muscle contractions circulate hemolymph in the appendages. Morphometric analysis shows that dendritic regression is similar in accessory planta retractor motoneurons with different pupal fates, and that accessory planta retractor motoneurons begin to participate in the pupal motor pattern while their dendrites are regressed. Accepted: 29 March 1998     

Dynamic behavior of excised dog rib cage: dependence on muscle

To assess the contribution of the rib cage to chest wall elastance and hysteresis, we measured force-displacement behavior of the isolated canine rib cage during sinusoidal forcing of the sternum in the midsagittal plane at low frequencies (0.02-2.0 Hz). Elastance of the rib cage was nearly invariant with frequency of forcing from 0.02 to 1.0 Hz and decreased with increasing amplitude. Hysteresis, the width of the force-displacement loop at middisplacement (zero displacement), was nearly constant with frequency below 1.0 Hz and increased with increasing amplitude of forcing. Removal of muscle reduced elastance and hysteresis of the rib cage substantially. The data suggest that the excised dog rib cage shows dynamic behavior similar to that of the intact human rib cage and chest wall and that respiratory muscle is responsible for a major part of the behavior of the passive chest wall. We also calculated the major and minor stiffnesses in the sagittal plane, which differed by a factor of 3-11, and their directions lay close to the dorsoventral and cephalocaudal axes, respectively. Removal of muscle reduced the stiffnesses but did not change their directions. Thus, although respiratory muscles impede motion in the sagittal plane, they do not alter its pattern.     

Neurogenic contractions of the rat tail artery under isobaric conditions: effect of transmural pressure and function of the endothelium]

In stimulation of the rat nerve with a modulated sine pattern, an increase in the modulating frequency from 0.03 to 0.15 Hz diminished the latency between the stimulating signals and changes in the vessel resistance as well as the amplitude of the flow oscillations, but did not affect tonic contractions of the vessel. A reduction of transmural pressure from 80 to 40 mm Hg increased both the tonic and the phasic components of the vessel contraction. Following the endothelium removal no change in the response latency occurred. The data obtained suggest that, during a rhythmic neurogenic influence, the vascular endothelium may work as an "amplifier" of the vessel's phasic contractions.     

Histochemical patterns in normal and splaylegged piglet muscle fibers

Summary The histochemical pattern of muscle fiber types of the longissimus dorsi and biceps femoris muscles was investigated in normal and splaylegged piglets at birth and seven days later. Only slight differences between the muscle fibers at birth were found using histochemical reactions for alkaline adenosine triphosphatase (ATPase), succinate dehydrogenase (SDH), phosphorylase (PH) activities, and for the periodic acid-Schiff (PAS) reaction. With the method for acid-preincubated ATPase activity, high activity was observed in Type I muscle fibers and low activity in Type II muscle fibers in animals of both groups investigated. However, a higher number of Type I fibers was found in muscles of normal piglets, suggesting a faster and more advanced process of transformation of Type II into Type I muscle fibers in unaffected animals. Thus the histochemical conversion appears to be retarded in muscles of splaylegged animals, which have a histochemical pattern similar to that of normal prenatal animals. Cholinesterase activity in motor endplates was well developed; its staining revealed smaller sized and irregularly arranged endplates in muscles of affected piglets. Fiber type differentiation in muscles of animals which recovered from splayleg becomes fully developed and comparable to normal piglets seven days after birth. The number of fibers which became converted from Type II to Type I was increased; the fiber types were differentiated with regard to the PAS reaction and to their ATPase, SDH and PH activities. Morphological features of motor endplates in muscles of normal and surviving splaylegged piglets are similar.Histochemical investigation of the fiber type differentiation thus suggests that full recovery occurs within the first week of postnatal life in muscles affected by pathological changes accompanying splayleg.     

Histochemical patterns in normal and splaylegged piglet muscle fibers

The histochemical pattern of muscle fiber types of the longissimus dorsi and biceps femoris muscles was investigated in normal and splaylegged piglets at birth and seven days later. Only slight differences between the muscle fibers at birth were found using histochemical reactions for alkaline adenosine triphosphatase (ATPase), succinate dehydrogenase (SDH), phosphorylase (PH) activities, and for the periodic acid-Schiff (PAS) reaction. With the method for acid-preincubated ATPase activity, high activity was observed in Type I muscle fibers and low activity in Type II muscle fibers in animals of both groups investigated. However, a higher number of Type I fibers was found in muscles of normal piglets, suggesting a faster and more advanced process of transformation of Type II into Type I muscle fibers in unaffected animals. Thus the histochemical conversion appears to be retarded in muscles of splaylegged animals, which have a histochemical pattern similar to that of normal prenatal animals. Cholinesterase activity in motor endplates was well developed; its staining revealed smaller sized and irregularly arranged endplates in muscles of affected piglets. Fiber type differentiation in muscles of animals which recovered from splayleg becomes fully developed and comparable to normal piglets seven days after birth. The number of fibers which became converted from Type II to Type I was increased; the fiber types were differentiated with regard to the PAS reaction and to their ATPase, SDH and PH activities. Morphological features of motor endplates in muscles of normal and surviving splaylegged piglets are similar. Histochemical investigation of the fiber type differentiation thus suggests that full recovery occurs within the first week of postnatal life in muscles affected by pathological changes accompanying splayleg.     

Dual action of intracellularly released calcium on the quantal mediator secretion

The mice diaphragm muscle and microelectrode technique were used to check the influence of ryanodine (0.5 mcM) on spontaneous and evoked mediator release under conditions of potassium depolarization (8-16 mM [K+]ex or rhythmic (4-100 Hz) stimulation of motor nerve terminals. Weak tonic calcium loading (by muscle exposition to 8 mM [K+]ex) caused a two-fold frequency increase if miniature and plate potentials (MEPPs), which was returned to the basal level by subsequent application of ryanodine. This inhibitory effect of ryanodine was blocked by apamin (500 nM) a blocker of K+(Ca)-channels. A greater calcium load of terminals (in solution with 16 mM [K+]ex) caused a 15-fold increase of MEPPs frequency. Subsequent ryanodine application caused an additional 2-3-fold increase of MEPPs frequency. During rhythmic activity of motor synapses, ryanodine was able to decrease the amplitude of EPP by 60% at plateau phase at short low frequency (4 Hz) of discharges and to increase the amplitude of EPP by 60-150% at high frequency (70-100 Hz) of discharges. It is concluded that rynodine induced calcium release from intraterminal Ca2+-stores can influence dual: excitatory or inhibitory, action on spontaneous and evoked mediator release, due to different intraterminal calcium loads and regimen of synaptic activity.     

Induction of a fatigue-resistant phenotype in rabbit fast muscle by small daily amounts of stimulation.

We have shown that fatigue resistance can be induced in rabbit tibialis anterior (TA) muscles without excessive power loss by continuous stimulation at low frequencies, such as 5 Hz, and that the same result is obtained by delivering a 10-Hz pattern in equal on/off periods. Here we ask whether the same phenotype could be produced with daily amounts of stimulation that would be more appropriate for clinical use. We stimulated rabbit TA muscles for 6 wk, alternating fixed 30-min on periods of stimulation at 10 Hz with off periods of different duration. All patterns transformed fast-glycolytic fibers into fast-oxidative fibers. The muscles had fatigue-resistant properties but retained a higher contractile speed and power production than muscles transformed completely to the slow-oxidative type. We conclude that in the rabbit as little as one 30-min period of stimulation in 24 h can result in a substantial increase in the resistance of the muscle to fatigue.     

Histochemical and stereological analysis of the levator ani (pubocaudal) muscle in nulliparous and multiparous beagles

Fiber-type composition and several stereological parameters of the levator ani (pubocaudal) muscle were evaluated in five nulliparous and five multiparous beagles using myosin ATPase-histochemistry and systematically selected muscle cross-sections. With respect to the narrow canine pelvic cavity, this study was also undertaken to determine whether vaginal birth of at least seven litters causes similar neuromuscular changes in the canine levator ani (pubocaudal) muscle analoguous to those seen in the pelvic floor muscles of women after vaginal delivery. The canine pubocaudal muscle is comprised of approximately equal amounts of slow twitch type I and fast twitch type II (IIA, IIS) fibers. The muscles of both the nulliparous and multiparous beagles did not display any signs indicative of denervation or myopathology. The multiparous dogs exhibited significantly increased mean absolute muscle (1720 mm(3)) and total fiber-type I volumes (850 mm(3)) as well as relevantly increased mean diameter of type I fibers (72.0 microm) when compared with the nulliparous group. The canine levator ani (pubocaudal) muscle is not pathologically affected by vaginal deliveries and seems to adapt to numerous successive pregnancies and births through fiber-type I hypertrophy.     

Effects of agonists and antagonists of rhyanodine receptors on potassium contractures in twitch and tonic frog skeletal muscle fibers

A comparative analysis of the effects of the concentrations of Ca2+ in external medium and the inhibitor (dantrolene) and activator (4-chloro-m-cresol) of rhyanodine-sensitive Ca2+ channels of carcoplasmic reticulum on the characteristics of potassium contracture in frog twitch and tonic skeletal muscles has been performed. It was shown that the duration of contracture in tonic muscles is not restricted by the presence of Ca2+, as distinct from twitch muscles. Dandrolene does not practically affect the contractile responses of tonic fibres, and the concentration of cresol eliciting the contracture for tonic fibres is substantially higher (1 mM) than for twitch fibers (0.25 mM). In twitch fibers, the potassium contracture activated in the presence of cresol is comparable in amplitude and dynamics with the contracture under control conditions, and in tonic fibers a summing of responses without relaxation after the washing of excessive potassium is observed. This suggests that, in twitch fibers, the influx of Ca2+ can directly create the concentration sufficient for the maintenance of contraction, and in tonic fibers its involvement is mediated through the Ca(2+)-dependent activation of the beta-isoform of rhyanodine-sensitive channels.