Locomotor muscles often perform diverse roles, functioning as motors that produce mechanical energy, struts that produce force and brakes that dissipate mechanical energy. In many vertebrate muscles, these functions are not mutually exclusive and a single muscle often performs a range of mechanically diverse tasks. This functional diversity has obscured the relationship between a muscle's locomotor function and its mechanical properties. I use hopping in toads as a model system for comparing muscles that primarily produce mechanical energy with muscles that primarily dissipate mechanical energy. During hopping, hindlimb muscles undergo active shortening to produce mechanical energy and propel the animal into the air, whereas the forelimb muscles undergo active lengthening to dissipate mechanical energy during landing. Muscles performing distinct mechanical functions operate on different regions of the force-length curve. These findings suggest that a muscle's operating length may be shaped by potential trade-offs between force production and sarcomere stability. In addition, the passive force-length properties of hindlimb and forelimb muscles vary, suggesting that passive stiffness functions to restrict the muscle's operating length in vivo. These results inform our understanding of vertebrate muscle variation by providing a clear link between a muscle's locomotor function and its mechanical properties.

Locomotor muscles often perform diverse roles, functioning as motors that produce mechanical energy, struts that produce force and brakes that dissipate mechanical energy. In many vertebrate muscles, these functions are not mutually exclusive and a single muscle often performs a range of mechanically diverse tasks. This functional diversity has obscured the relationship between a muscle's locomotor function and its mechanical properties. I use hopping in toads as a model system for comparing muscles that primarily produce mechanical energy with muscles that primarily dissipate mechanical energy. During hopping, hindlimb muscles undergo active shortening to produce mechanical energy and propel the animal into the air, whereas the forelimb muscles undergo active lengthening to dissipate mechanical energy during landing. Muscles performing distinct mechanical functions operate on different regions of the force-length curve. These findings suggest that a muscle's operating length may be shaped by potential trade-offs between force production and sarcomere stability. In addition, the passive force-length properties of hindlimb and forelimb muscles vary, suggesting that passive stiffness functions to restrict the muscle's operating length in vivo. These results inform our understanding of vertebrate muscle variation by providing a clear link between a muscle's locomotor function and its mechanical properties.

Skeletal muscle exhibits broad functional diversity, despite its inherent length and velocity constraints. The observed variation in morphology and physiology is assumed to have evolved to allow muscle to operate at its optimal length and velocity during locomotion. Here, we used the variation in optimum lengths and velocities that occurs with muscle activation level to experimentally test this assumption. Muscle ergometry and sonomicrometry were used to characterize force-length and power-velocity relationships, and in vivo operating lengths and velocities, at a range of activation levels. Operating lengths and velocities were mapped onto activation level specific force-length and power-velocity relationships to determine whether they tracked changing optima. Operating velocities decreased in line with decreased optimal velocities, suggesting that optimal velocities are always used. However, operating lengths did not change with changing optima. At high activation levels, fibres used an optimal range of lengths. However, at lower activation levels, fibres appeared to operate on the ascending limb of sub-maximally activated force-length relationships. This suggests that optimal lengths are only used when demand is greatest. This study provides the first mapping of operating lengths to activation level-specific optima, and as such, provides insight into our assumptions about the factors that determine muscle performance during locomotion.

Anurans (frogs and toads) have been shown to have relatively compliant skeletal muscles. Using a meta-analysis of published data we have found that muscle stiffness is negatively correlated with joint range of motion when examined across mammalian, anuran and bird species. Given this trend across a broad phylogenetic sample, we examined whether the relationship held true within anurans. We identified four species that differ in preferred locomotor mode and hence joint range of motion (Lithobates catesbeianus, Rhinella marina, Xenopus laevis and Kassina senegalensis) and hypothesized that smaller in vivo angles (more flexed) at the knee and ankle joint would be associated with more compliant extensor muscles. We measured passive muscle tension during cyclical stretching (20%) around L0 (sarcomere lengths of 2.2μm) in fiber bundles extracted from cruralis and plantaris muscles. We found no relationship between muscle stiffness and range of motion for either muscle-joint complex. There were no differences in the passive properties of the cruralis muscle among the four species, but the plantaris muscles of the Xenopus and Kassina were significantly stiffer than those of the other two species. Our results suggest that in anurans the stiffness of muscle fibers is a relatively minor contributor to stiffness at the level of joints and that variation in other anatomical properties including muscle-tendon architecture and joint mechanics as well as active control likely contribute more significantly to range of motion during locomotion.

The force, mechanical work and power produced by muscle fibers are profoundly affected by the length changes they undergo during a contraction. These length changes are in turn affected by the spatial orientation of muscle fibers within a muscle (fiber architecture). Therefore any heterogeneity in fiber architecture within a single muscle has the potential to cause spatial variation in fiber strain. Here we examine how the architectural variation within a pennate muscle and within a fusiform muscle can result in regional fiber strain heterogeneity. We combine simple geometric models with empirical measures of fiber strain to better understand the effect of architecture on fiber strain heterogeneity. We show that variation in pennation angle throughout a muscle can result in differences in fiber strain with higher strains being observed at lower angles of pennation. We also show that in fusiform muscles, the outer/superficial fibers of the muscle experience lower strains than central fibers. These results show that regional variation in mechanical output of muscle fibers can arise solely from architectural features of the muscle without the presence of any spatial variation in motor recruitment.

Skeletal muscles are rarely recruited maximally during movement. However, much of our understanding of muscle properties is based on studies using maximal activation. The effect of activation level on skeletal muscle properties remains poorly understood. Muscle optimum length increases with decreased activation; however, the mechanism responsible is unclear. Here, we attempted to determine whether length-dependent calcium effects, or the effect of absolute force underpin this shift. Fixed-end contractions were performed in frog plantaris muscles at a range of lengths using maximal tetanic (high force, high calcium), submaximal tetanic (low force, high calcium) and twitch (low force, low calcium) stimulation conditions. Peak force and optimum length were determined in each condition. Optimum length increased with decreasing peak force, irrespective of stimulation condition. Assuming calcium concentration varied as predicted, this suggests that absolute force, rather than calcium concentration, underpins the effect of activation level on optimum length. We suggest that the effect of absolute force is due to the varying effect of the internal mechanics of the muscle at different activation levels. These findings have implications for our understanding of in vivo muscle function and suggest that mechanical interactions within muscle may be important determinants of force at lower levels of activation.

The collagenous extracellular matrix (ECM) of skeletal muscle functions to transmit force, protect sensitive structures, and generate passive tension to resist stretch. The mechanical properties of the ECM change with age, atrophy, and neuromuscular pathologies, resulting in an increase in the relative amount of collagen and an increase in stiffness. Although numerous studies have focused on the effect of muscle fibrosis on passive muscle stiffness, few have examined how these structural changes may compromise contractile performance. Here we combine a mathematical model and experimental manipulations to examine how changes in the mechanical properties of the ECM constrain the ability of muscle fibers and fascicles to radially expand and how such a constraint may limit active muscle shortening. We model the mechanical interaction between a contracting muscle and the ECM using a constant volume, pressurized, fiber-wound cylinder. Our model shows that as the proportion of a muscle cross section made up of ECM increases, the muscle's ability to expand radially is compromised, which in turn restricts muscle shortening. In our experiments, we use a physical constraint placed around the muscle to restrict radial expansion during a contraction. Our experimental results are consistent with model predictions and show that muscles restricted from radial expansion undergo less shortening and generate less mechanical work under identical loads and stimulation conditions. This work highlights the intimate mechanical interaction between contractile and connective tissue structures within skeletal muscle and shows how a deviation from a healthy, well-tuned relationship can compromise performance.