Isometric contractile properties and instantaneous stiffness of amphibian skeletal muscle in the temperature range from 0 to 20 degrees C

The isometric contractile properties of frog (Rana pipiens) and toad (Bufo bufo) sartorii have been studied over the temperature range from 0 to 20 degrees C. The isometric twitch tension was found to vary considerably between these two species and between muscles in the same species. Between 0 and 4 degrees C there was very little change in maximum isometric twitch tension. Between 4 and 12 degrees C several muscles from frog or toad showed a potentiation of twitch tension whereas others showed a decline. Over this temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature range the toad sartorii consistently demonstrated a greater potentiation. By 12 degrees C a steady decline in twitch tension in both muscles was seen as the temperature approached 20 degrees C. The maximum isometric tetanic tension recorded between 18 and 20 degrees C increased fractionally to an average of 1.504 +/- 0.029 (n = 4) for frog sartorii and to 1.377 +/- 0.008 (n = 5) for toad sartorii. The time to peak twitch tension and the half-relaxation time decreased markedly with an increase in temperature. Moreover, the half-relaxation time was reduced by a greater proportion than the time to peak twitch tension. Measurements of instantaneous stiffness by controlled velocity releases from the plateau of isometric tetani revealed that the large increase in isometric tetanus tension as the muscle was warmed was not accompanied by a corresponding increase in the total number of active cross-bridges. The possibility that a decreased availability of intracellular Ca2+ ions at the contractile sites contributing to the fall of isometric twitch tension at elevated temperatures is discussed. The possibility exists that at elevated temperatures a change inthe intrinsic contractile ability of the muscle occurs which produces an increased tension per cross-bridge.     

Properties of twitch motor units of the ankle extensor muscles in the bullfrog Rana catesbeiana

A study of the mechanical properties of the twitch motor units in the ankle extensor muscles of bullfrogs was undertaken to expand our view of the diversity of motor unit properties among vertebrates. Two muscles were chosen that represent a wide range of extensor function: the plantaris longus (PL) is a large muscle providing most of the force for ankle extension in hopping and swimming, and the tibialis posticus (TP) is relatively small and may act as an ankle stabilizer or be primarily postural in function. Both muscles have highly fatigable motor units, but also some (especially in TP) low or non-fatigable ones. Mean tetanic tensions of motor units in both muscles are relatively large as compared with those of mammals but are especially large in PL, No clear correlations were found between contraction times and either motor unit tetanic tensions or fatigability, nor did the motor units fall into clearly defined types based on any functional parameters. Overall contraction and relaxation times are slow compared with those of mammals and are somewhat slow compared to those of other frogs; unlike results from earlier studies, the large units of PL are slower than the small units of TP. This results in PL units reaching fused tetani at lower stimulus frequencies. The twitch/tetanus and force/frequency ratios in PL motor units are much larger than those of TP, giving PL units greater relative forces at lower stimulus rates. These results are discussed in the context of motor unit function. © 1994 Wiley-Liss, Inc.     

Hindlimb muscle fiber types in two frogs (Rana catesbeiana and Litoria caerulea) with different locomotor behaviors: histochemical and enzymatic comparison

To test how differences in locomotor behaviors may be reflected in muscle fiber-type diversity within anurans, a comparison of hindlimb muscles between the powerful terrestrial hopper, Rana catesbeiana, and the tree frog, Litoria caerulea, was done. One postural muscle (tibialis posticus, TP) and one primary hopping muscle (plantaris longus, PL), were characterized to identify muscle fiber types using standard histochemical methods. In addition, spectophotometric analysis of activity levels of the oxidative enzyme citrate synthase (CS) and the glycolytic enzyme lactate dehydrogenase (LDH) were done in each muscle. In spite of presumed differences in behavior between the species, we found no significant differences in the proportions of the identified fiber types when the muscles were compared across species. In addition, there were no significant differences in the proportions of the different fiber types between the postural versus phasic muscles within species. Within Rana, the postural muscle (TP) had greater oxidative capacity (as measured by CS activity) than did the phasic muscle (PL). Both muscles had equivalent LDH activities. Within Litoria, PL and TP did not differ in either LDH or CS activities. Both PL and TP of Litoria had less LDH activity and greater CS activity than their homologs in Rana. Thus, in spite of the uniform populations of fiber types between muscles and species, the metabolic diversity based on enzyme activity is consistent with behavioral differences between the species. These results suggest that the range of functional diversity within fiber types may be very broad in anurans, and histochemical fiber typing alone is not a clear indicator of their metabolic or functional properties.     

Morphometric relationships of take-off speed in anuran amphibians

Locomotory speed correlates with muscle mass (determining force and stride rate), limb length (stride rate and distance), and laterally compressed body trunk (force and stride distance). To delineate generalization of the locomotory-morphometric relationships specifically in anuran amphibians, we investigated take-off speed and the three morphological variables from seven species, Rana nigromaculata, R. rugosa, and Bombina orientalis, Eleuthrodectilus fitzingeri, E. diastema, Bufo typhonius, Colostethus flotator and Physalaemus pustulosus. The fastest jumper E. fitzingeri (3.41 m s(-1)) showed 2.49-fold greater speed than the slowest B. typhonius. Take-off speed correlated well with both thigh muscle mass relative to body mass and hindlimb length relative to snout-vent length (HL/SVL), but poorly correlated with the inter-ilial width relative to SVL. The best morphological predictor was HL/SVL (speed=-3.28+3.916 HL/SVL, r=0.968, P<0.0001), suggesting that anuran take-off speed is portrayed well with high gear and acceleration distance characterized by hindlimbs.     

Preparation and characterization of the major isotype of parvalbumin from skeletal muscle of the toad (Bufo bufo japonicus)

The major isotype of parvalbumin has been isolated from the skeletal muscle of the toad, Bufo bufo japonicus. Unlike the skeletal muscle of every frog so far examined (Rana esculenta, Rana temporaria, and Rana catesbeiana), which contains two major isotypes of parvalbumins, toad skeletal muscle has been shown to contain only one isotype, but the content of parvalbumin in toad skeletal muscle was similar to the sum of those of the two isotypes in skeletal muscles of frogs. This feature of toad skeletal muscle is advantageous to clarify the physiological role of parvalbumin. The relative molecular mass of toad parvalbumin was estimated to be 12,200 by SDS-polyacrylamide gel electrophoresis. The isoelectric point was determined to be 4.81 by polyacrylamide gel isoelectric focusing. The amino acid composition indicated that toad parvalbumin corresponds to bullfrog (R. catesbeiana) pI 4.97 parvalbumin, showing that toad parvalbumin is genetically an alpha-parvalbumin. It was also revealed by the amino acid composition that toad parvalbumin is distinctly different from any of the parvalbumins from frogs. The ultraviolet spectrum of toad parvalbumin is consistent with its amino acid composition. The ultraviolet difference spectrum of the Ca2+-loaded form vs. the metal-free form indicates that some Phe residues in the toad parvalbumin molecule are affected by a conformational change associated with Ca2+ binding. On electrophoresis in polyacrylamide gel in 14 mM Tris and 90 mM glycine, the metal-free and Mg2+-loaded forms of toad parvalbumin migrated twice as fast as the Ca2+-loaded form.(ABSTRACT TRUNCATED AT 250 WORDS)     

Breeding habitat selection and larval performance of two anurans in freshwater rock-pools

I studied breeding habitat choice of common toad Bufo bufo and common frog Rana temporaria in 221 freshwater rock-pools on 15 small islands in the southwestern coast of Finland. I monitored tadpole growth rate in a number of rock-pools differing in size, competitors and predator regime. Furthermore, I carried out a short-term field experiment to investigate the effects of different predators on larval survival. Rana bred in pools of virtually all sizes, although the very smallest ones were usually avoided. A logistic regression analysis showed that the amount of aquatic macrophyte vegetation and pool surface area were the most important pool characters for Rana. Bufo preferred the largest pools, and pool surface area was by far the most important habitat variable. Rana avoided pools with the lowest pH. but was more common than expected at pools with pH around 6. In Bufo such an effect was not found, but this may be due to relatively low number of pools occupied by Bufo. In Rana there was a positive correlation between pool area and tadpole body length in early June, but two weeks later this relationship was not significant. In Bufo tadpole body size was not significantly affected by pool area. Presence of predators or presence of the other tadpole species did not affect growth or tadpole body size in either Rana or Bufo. A short-term experiment in wading pools indicated that both species suffered heavy mortality in the presence of a diving beetle larva. In the presence of predatory fish, mortality of Bufo did not differ from controls, whereas mortality was high among Rana tadpoles. It appears that although predators prey heavily on Rana tadpoles, they do not exclude Rana from larger rock-pools. However, it is not evident from this study why Bufo avoids smaller pools.     

Skull allometry in the marine toad,Bufo marinus

Scaling predictions pioneered by A.V. Hill state that isometric changes in kinematics result from isometric changes in size. These predictions have been difficult to support because few animals display truly isometric growth. An exception to this rule is said to be the toads in the genus Bufo, which can grow over three orders of magnitude. To determine whether skull shape increases isometrically, I used linear measurements and geometric morphometrics to quantify shape variation in a size series of 69 skulls from the marine toad, B. marinus. Toads ranged in body mass from 1.8 gm to a calculated 1,558.9 gm. Of all linear measurements (S/V length, skull width, skull length, levator mass, depressor mass, adductor foramen area), only the area of the adductor foramen increased faster than body mass; the remaining variables increased more slowly. In addition, modeling the lower jaw as a lever‐arm system showed that the lengths of the closing in‐ and out‐levers scaled isometrically with body mass despite the fact that the skull itself is changing allometrically. Geometric morphometrics discerned areas of greatest variability with increasing body mass at the rear of the skull in the area of the squamosal bone and the adductor foramen. This increase in area of the adductor foramen may allow more muscle to move the relatively greater mass of the lower jaw in larger toads, although adductor mass scales with body mass. If B. marinus feeds in a similar manner to other Bufo, these results imply that morphological allometry may still result in kinematic isometry. J. Morphol. 241:115–126, 1999. © 1999 Wiley‐Liss, Inc.     

Effect of temperature and Zn2+ on isometric contractile properties and electrical phenomena of frog (Rana) and Xenopus skeletal muscle fibers

Effects of temperature and Zn2+ on the isometric contractile properties of toe muscle fibers of Rana catesbeiana and Xenopus laevis were studied. The maximum twitch tension almost doubled when the temperature was lowered from 20 to 4 degrees C in Rana muscles but not in Xenopus muscles, although the duration of action potential in Xenopus muscle was increased slightly more than that seen in the Rana species. The maximum rate of rise of tension was greater in Xenopus muscle than in the Rana muscle, at 20 degrees C. The prolongation of the time-to-peak tension following exposure to low temperature (4 degrees C) was more pronounced in Rana than in Xenopus muscles. These results suggest that the speed of release and reuptake of Ca2+ by the sarcoplasmic reticulum (SR) differs in Rana and Xenopus muscles and that these factors may be related to differences in the SR and the T-tubular morphology. In Rana muscles, Zn2+ prolonged the falling phase of the action potential and potentiated the twitch tension. In Xenopus muscles, Zn2+ marginally prolonged the duration of action potential and the twitch tension was not markedly potentiated. These results indicate that Zn2+ potentiates the twitch by prolonging the action potential and that Rana muscles are more sensitive to the effects of Zn2+.     

Coexistence of twitch potentiation and tetanic force decline in rat hindlimb muscle

An experimental protocol designed to assess fatigability in motor units has been applied to two hindlimb muscles of anesthetized adult rats to study the effects of whole-muscle fatigue on the isometric twitch. Both soleus and extensor digitorum longus exhibited a linear relationship between fatigability (i.e., force decline after a 360-s fatigue test) and the magnitude of the twitch force following the fatigue test. Twitch force after the fatigue test was potentiated (i.e., greater than the value before the fatigue test) in many muscles, despite the development of considerable fatigue. This coexistence of fatigue and twitch potentiation was observed in 7% (5/70) of soleus and 48% (31/64) of extensor digitorum longus muscles. The coexistence was exhibited only by the least fatigable muscles of the fast-contracting extensor digitorum longus. The extensor digitorum longus muscles that did not exhibit twitch potentiation probably experienced a higher proportion of muscle-fiber inactivation, such as due to failure of neuromuscular propagation, that was induced by the fatigue regimen.     

Blood flow and glycogen use in hypertrophied rat muscles during exercise

Previous findings suggest that skeletal muscle that has enlarged as a result of removal of synergistic muscles has a similar metabolic capacity and improved resistance to fatigue compared with normal muscle. The purpose of the present study was to follow blood flow and glycogen loss patterns in hypertrophied rat plantaris plantaris and soleus muscles during treadmill exercise to provide information on the adequacy of perfusion of the muscles during in vivo exercise. Thirty days following surgical removal of gastrocnemius muscle, blood flows (determined with radiolabeled microspheres) and glycogen concentrations were determined in all of the ankle extensor muscles of experimental and sham-operated control rats during preexercise and after 5-6 min of treadmill exercise at 15 m/min. There were no differences (P greater than 0.05) in blood flows per unit mass or glycogen concentrations between control and hypertrophied plantaris or soleus muscles at either time, although both muscles were larger (P less than 0.05) in the experimental group (plantaris: 95%; soleus: 40%). None of the other secondary ankle extensor muscles (tibialis posterior, flexor digitorum longus or flexor hallicus longus) hypertrophied in response to removal of gastrocnemius. These results provide indirect evidence that O2 delivery in the enlarged muscles is not compromised during low-intensity treadmill exercise due to limited perfusion.     

Inward rectification in muscle fibres of the toad Bufo marinus

Inward rectification of the resting potassium conductance was studied in skeletal muscle fibres of the toad Bufo marinus. This conductance was shown to be blocked by Ba and Cs and located both in the surface membrane and the membranes of the tubular system. Some differences were found between the properties of this conductance channel in Bufo marinus and those reported for various species of Rana. The possible adaptative value of these differences is pointed out.     

Relation between the ankle joint angle and the maximum isometric force of the toe flexor muscles

The purpose of this study was to investigate the relationships between the ankle joint angle and maximum isometric force of the toe flexor muscles. Toe flexor strength and electromyography activity of the foot muscles were measured in 12 healthy men at 6 different ankle joint angles with the knee joint at 90 deg in the sitting position. To measure the maximum isometric force of the toe flexor muscles, subjects exerted maximum force on a toe grip dynamometer while the activity levels of the intrinsic and extrinsic plantar muscles were measured. The relation between ankle joint angle and maximum isometric force of the toe flexor muscles was determined, and the isometric force exhibited a peak when the ankle joint was at 70–90 deg on average. From this optimal neutral position, the isometric force gradually decreased and reached its nadir in the plantar flexion position (i.e., 120 deg). The EMG activity of the abductor hallucis (intrinsic plantar muscle) and peroneus longus (extrinsic plantar muscle) did not differ at any ankle joint angles. The results of this study suggest that the force generation of toe flexor muscles is regulated at the ankle joint and that changes in the length-tension relations of the extrinsic plantar muscle could be a reason for the force-generating capacity at the metatarsophalangeal joint when the ankle joint angle is changed.     

Older adults are less steady during submaximal isometric contractions with the knee extensor muscles.

This study compared the steadiness of submaximal contractions with the knee extensor muscles in young and old adults. Twenty young and twenty old subjects underwent assessment of isometric maximum voluntary contraction (MVC), one-repetition maximum (1-RM) strength, and steadiness during isometric, concentric, and eccentric contractions with the knee extensor muscles. The old adults displayed 33% lower MVC force and a 41% lower 1-RM load. The coefficient of variation for force was significantly greater for the old adults during isometric contractions at 2, 5, and 10% of MVC but not at 50% MVC. The decline in steadiness at low forces experienced by the men was marginally greater than that experienced by the women. The steadiness of concentric and eccentric contractions was similar in young and old adults at 5, 10, and 50% of 1-RM load. Old subjects exhibited greater coactivation of an antagonist muscle compared with young subjects during the submaximal isometric and anisometric contractions. These results indicate that, whereas the ability to exert steady submaximal forces with the knee extensor muscles was reduced in old adults, fluctuations in knee joint angle during slow movements were similar for young and old adults.     

Lower extremity control and dynamics during backward angular impulse generation in forward translating tasks

Observation of complex whole body movements suggests that the nervous system coordinates multiple operational subsystems using some type of hierarchical control. When comparing two forward translating tasks performed with and without backward angular impulse, we have learned that both trunk-leg coordination and reaction force-time characteristics are significantly different between tasks. This led us to hypothesize that differences in trunk-leg coordination and reaction force generation would induce between-task differences in the control of the lower extremity joints during impulse generation phase of the tasks. Eight highly skilled performers executed a series of forward jumps with and without backward rotation (reverse somersault and reverse timer, respectively). Sagittal plane kinematics, reaction forces, and electromyograms of lower extremity muscles were acquired during the take-off phase of both tasks. Lower extremity joint kinetics were calculated using inverse dynamics. The results demonstrated between-task differences in the relative angles between the lower extremity segments and the net joint forces/reaction force and the joint angular velocity profiles. Significantly less knee extensor net joint moments and net joint moment work and greater hip extensor net joint moments and net joint moment work were observed during the push interval of the reverse somersault as compared to the reverse timer. Between-task differences in lower extremity joint kinetics were regulated by selectively activating the bi-articular muscles crossing the knee and hip. These results indicate that between-task differences in the control of the center of mass relative to the reaction force alters control and dynamics of the multijoint lower extremity subsystem.     

Hopping and swimming in the leopard frog Rana pipiens: II. A comparison of muscle activities

Electromyography (EMG) was used to examine muscle activity of the major hip, knee, and ankle extensors during both hopping and swimming in leopard frogs. Chronic EMG electrodes were implanted for periods of 7–10 days. This permitted us to record EMG activities during both hopping and swimming from the same electrode, allowing a direct comparison of the timing and amplitudes of muscle activity between the two behaviors. We could then relate these activities to the kinematics of locomotion. In both behaviors, all three extensors were synchronously activated 30–50 ms before limb extension began. However, the hip extensor turned on relatively earlier in hopping than in swimming when on time was expressed as percent of stride. The hip and knee extensors were activated relatively longer in hopping and the ankle extensor relatively longer in swimming. The amplitudes of the rectified, integrated EMG signals were roughly twice as large in hopping as in swimming for all three muscles, supporting the notion that propulsion in hopping requires more force than in swimming. The EMG burst durations differed little between the muscles or, in relative duration, between the behaviors. As has been found in other quadrupeds, the EMG bursts began before visible movement and ceased at or before hindlimb extension was completed. In our animals, however, we found a consistent, low level (10–30% of maximum amplitude) of EMG activity that continued 60–200 ms past the end of the burst and into the suspension periods in both hopping and swimming. We hypothesize that this unusual activity may be present in frogs so that the hind limb remains aero(hydro)dynamically stable as the frog arches through its leap or glides in swimming following completed limb extension. Thus, the timing and pattern of the EMG bursts are consistent with those present in other tetrapods and support conservatism of neural control. However, the prolonged low-level activity suggests flexibility in the control pattern and variation according to specific behaviors. © 1996 Wiley-Liss, Inc.     

Strength training counteracts motor performance losses during bed rest.

The purpose of the study was to determine the effect of bed rest with or without strength training on torque fluctuations and activation strategy of the muscles. Twelve young men participated in a 20-day bed rest study. Subjects were divided into a non-training group (BRCon) and a strength-training group (BRTr). The training comprised dynamic calf-raise and leg-press exercises. Before and after bed rest, subjects performed maximal contractions and steady submaximal isometric contractions of the ankle extensor muscles and of the knee extensor muscles (2.5-10% of maximal torque). Maximal torque decreased for both the ankle extensors (9%, P < 0.05) and knee extensors (16%, P < 0.05) in BRCon but not in BRTr. For the ankle extensors, the coefficient of variation (CV) for torque increased in both groups (P < 0.05), with a greater amount (P < 0.05) in BRCon (88%) compared with BRTr (41%). For the knee extensors, an increase in the CV for torque was observed only in BRCon (22%). The increase in the CV for torque in BRCon accompanied the greater changes in electromyogram amplitude of medial gastrocnemius (122%) and vastus lateralis (59%) compared with BRTr (P < 0.05). The results indicate that fluctuations in torque during submaximal contractions of the extensor muscles in the leg increase after bed rest and that strength training counteracted the decline in performance. The response varied across muscle groups. Alterations in muscle activation may lead to an increase in fluctuations in motor output after bed rest.     

The tactile-stimulated startle response of tadpoles: acceleration performance and its relationship to the anatomy of wood frog (Rana sylvatica), bullfrog (Rana catesbeiana), and American toad (Bufo americanus) tadpoles

I described the tactile-stimulated startle response (TSR) of wood frog (Rana sylvatica), bullfrog (Rana catesbeiana), and American toad (Bufo americanus) tadpoles. One purpose was to rank species in terms of maximum acceleration performance. Also, I tested whether anatomical indicators of performance potential were predictive of realized performance. TSRs were elicited in a laboratory setting, filmed at 250 Hz, and digitally analyzed. TSRs began with two, initial body curls during which tadpoles showed a broad spectrum of movement patterns. TSR performance was quantified by maximum linear acceleration and maximum rotational acceleration of the head/body, both of which tended to occur immediately upon initiation of motion (< 0.012 sec into the response). Bullfrog tadpoles had higher maximum acceleration than the other species, but other interspecific differences were not significant. The species' rank order for the anatomical indicator of linear acceleration potential was bullfrog > wood frog > American toad. The species' rank order for the anatomical indicator of rotational acceleration potential was bullfrog > wood frog = American toad. Thus, the anatomical indicators roughly predicted the rank order of interspecific average performance. However, the anatomical indicators did not correlate with individual tadpole performance. Variability in behavioral patterns may obscure the connection between anatomy and performance. This is seen in the current lack of intraspecific correlation between a morphological indicator of acceleration capacity and acceleration performance.     

Differential design for hopping in two species of wallabies.

Hindlimb musculoskeletal anatomy and steady speed over ground hopping mechanics were compared in two species of macropod marsupials, tammar wallabies and yellow-footed rock wallabies (YFRW). These two species are relatively closely related and are of similar size and general body plan, yet they inhabit different environments with presumably different musculoskeletal demands. Tammar wallabies live in relatively flat, open habitat whereas yellow-footed rock wallabies inhabit steep cliff faces. The goal of this study was to explore musculoskeletal differences between tammar wallabies and yellow-footed rock wallabies and determine how these differences influence each species' hopping mechanics. We found the cross-sectional area of the combined ankle extensor tendons of yellow-footed rock wallabies was 13% greater than that of tammar wallabies. Both species experienced similar ankle joint moments during steady-speed hopping, however due to a lower mechanical advantage at this joint, tammar wallabies produced 26% more muscle force. Thus, during moderate speed hopping, yellow-footed rock wallabies operated with 38% higher tendon safety factors, while tammar wallabies were able to store 73% more elastic strain energy (2.18 J per leg vs. 1.26 J in YFRW). This likely reflects the differing demands of the environments inhabited by these two species, where selection for non-steady locomotor performance in rocky terrain likely requires trade-offs in locomotor economy.     

Adjustment of muscle mechanics model parameters to simulate dynamic contractions in older adults

The generation of muscle-actuated simulations that accurately represent the movement of old adults requires a model that accounts for changes in muscle properties that occur with aging. An objective of this study was to adjust the parameters of Hill-type musculo-tendon models to reflect nominal age-related changes in muscle mechanics that have been reported in the literature. A second objective was to determine whether using the parametric adjustments resulted in simulated dynamic ankle torque behavior similar to that seen in healthy old adults. The primary parameter adjustment involved decreasing maximum isometric muscle forces to account for the loss of muscle mass and specific strength with age. A review of the literature suggested the need for other modest adjustments that account for prolonged muscular deactivation, a reduction in maximum contraction velocity, greater passive muscle stiffness and increased normalized force capacity during lengthening contractions. With age-related changes incorporated, a musculo-tendon model was used to simulate isometric and isokinetic contractions of ankle plantarflexor and dorsiflexor muscles. The model predicted that ankle plantarflexion power output during 120 deg/s shortening contractions would be over 40% lower in old adults compared to healthy young adults. These power losses with age exceed the 30% loss in isometric strength assumed in the model but are comparable to 39-44% reductions in ankle power outputs measured in healthy old adults of approximately 70 years of age. Thus, accounting for age-related changes in muscle properties, other than decreased maximum isometric force, may be particularly important when simulating movements that require substantial power development.