Evolution of a high-performance and functionally robust musculoskeletal system in salamanders.

The evolution of ballistic tongue projection in plethodontid salamanders-a high-performance and thermally robust musculoskeletal system-is ideal for examining how the components required for extreme performance in animal movement are assembled in evolution. Our comparative data on whole-organism performance measured across a range of temperatures and the musculoskeletal morphology of the tongue apparatus were examined in a phylogenetic framework and combined with data on muscle contractile physiology and neural control. Our analysis reveals that relatively minor evolutionary changes in morphology and neural control have transformed a muscle-powered system with modest performance and high thermal sensitivity into a spring-powered system with extreme performance and functional robustness in the face of evolutionarily conserved muscle contractile physiology. Furthermore, these changes have occurred in parallel in both major clades of this largest family of salamanders. We also find that high-performance tongue projection that exceeds available muscle power and thermal robustness of performance coevolve, both being emergent properties of the same elastic-recoil mechanism. Among the taxa examined, we find muscle-powered and fully fledged elastic systems with enormous performance differences, but no intermediate forms, suggesting that incipient elastic mechanisms do not persist in evolutionary time. A growing body of data from other elastic systems suggests that similar coevolution of traits may be found in other ectothermic animals with high performance, particularly those for which thermoregulation is challenging or ecologically costly.

Movements of vastly different performance have similar underlying muscle physiology.

Many animals use elastic recoil mechanisms to power extreme movements, achieving levels of performance that would not be possible using muscle power alone. Contractile performance of vertebrate muscle depends strongly on temperature, but the release of energy from elastic structures is far less thermally dependent, thus elastic recoil confers thermal robustness to whole-animal performance. Here we explore the role that muscle contractile properties play in the differences in performance and thermal robustness between elastic and non-elastic systems by examining muscle from two species of plethodontid salamanders that use elastically powered tongue projection to capture prey and one that uses non-elastic tongue projection. In species with elastic mechanisms, tongue projection is characterized by higher mechanical power output and thermal robustness compared with tongue projection of closely related genera with non-elastic mechanisms. In vitro and in situ muscle experiments reveal that species differ in their muscle contractile properties, but these patterns do not predict the performance differences between elastic and non-elastic tongue projection. Overall, salamander tongue muscles are similar to other vertebrate muscles in contractile performance and thermal sensitivity. We conclude that changes in the tongue-projection mechanism, specifically the elaboration of elastic structures, are responsible for high performance and thermal robustness in species with elastic tongue projection. This suggests that the evolution of high-performance and thermally robust elastic recoil mechanisms can occur via relatively simple changes to morphology, while muscle contractile properties remain relatively unchanged.

Thermal sensitivity of motor control of muscle-powered versus elastically powered tongue projection in salamanders.

Elastic-recoil mechanisms can improve organismal performance and circumvent the thermal limitations of muscle contraction, yet they require the appropriate motor control to operate. We compare muscle activity during tongue projection in salamanders with elastically powered, ballistic projection with activity of those with muscle-powered, non-ballistic projection across a range of temperatures to understand how motor control is integrated with elastically powered movements, and how this integration contributes to reduced thermal sensitivity. Species with ballistic tongue projection activated and deactivated their projector muscles significantly earlier than non-ballistic species, in a pattern consistent with a mechanism in which the muscle strains elastic tissue that subsequently recoils to power projection. Tongue projection was more thermally robust in ballistic species, but in both ballistic and non-ballistic species the projector muscles were activated earlier and for longer as temperature decreased. The retractor muscles showed a pattern similar to that of the projector muscles, but declined in a similar manner in the two groups. Muscle activity intensity also decreased at low temperatures in both groups, revealing that compensatory muscle activation does not account for the improved thermal robustness in ballistic species. Thus, relatively minor shifts in motor patterns accompanying morphological changes such as increased elastic tissue are sufficient to improve performance and decrease its thermal sensitivity without specialization of muscle contractile physiology.

Extreme Performance and Functional Robustness of Movement are Linked to Muscle Architecture: Comparing Elastic and Nonelastic Feeding Movements in Salamanders.

Muscle-powered movements are limited by the contractile properties of muscles and are sensitive to temperature changes. Elastic-recoil mechanisms can both increase performance and mitigate the effects of temperature on performance. Here, we compare feeding movements in two species of plethodontid salamanders, Bolitoglossa franklini and Desmognathus quadramaculatus, across a range of body temperatures (5-25°C) to better understand the mechanism of elastically powered, thermally robust movements. Bolitoglossa exhibited ballistic, elastically powered tongue projection with a maximum muscle mass specific power of 4,642 W kg(-1) while Desmognathus demonstrated nonballistic, muscle-powered tongue projection with a maximum power of 359 W kg(-1) . Tongue-projection performance in Bolitoglossa was more thermally robust than that of Desmognathus, especially below 15°C. The improved performance and thermal robustness of Bolitoglossa was associated with morphological changes in the projector muscle, including elaborated collagen aponeuroses and the absence of myofibers attaching directly to the tongue skeleton. The elongated aponeuroses likely increase the capacity for elastic energy storage, and the lack of myofibers inserting on the tongue skeleton permits ballistic projection. These results suggest that relatively simple changes in myofiber architecture and the amount of connective tissue can improve the performance and functional robustness of movements in the face of environmental challenges such as variable temperature.

Adaptive evolution in locomotor performance: How selective pressures and functional relationships produce diversity.

Despite the complexity of nature, most comparative studies of phenotypic evolution consider selective pressures in isolation. When competing pressures operate on the same system, it is commonly expected that trade-offs will occur that will limit the evolution of phenotypic diversity, however, it is possible that interactions among selective pressures may promote diversity instead. We explored the evolution of locomotor performance in lizards in relation to possible selective pressures using the Ornstein-Uhlenbeck process. Here, we show that a combination of selection based on foraging mode and predator escape is required to explain variation in performance phenotypes. Surprisingly, habitat use contributed little explanatory power. We find that it is possible to evolve very different abilities in performance which were previously thought to be tightly correlated, supporting a growing literature that explores the many-to-one mapping of morphological design. Although we generally find the expected trade-off between maximal exertion and speed, this relationship surprisingly disappears when species experience selection for both performance types. We conclude that functional integration need not limit adaptive potential, and that an integrative approach considering multiple major influences on a phenotype allows a more complete understanding of adaptation and the evolution of diversity.

Dynamics and thermal sensitivity of ballistic and non-ballistic feeding in salamanders.

Low temperature reduces the performance of muscle-powered movements, but in movements powered by elastic recoil mechanisms, this effect can be mitigated and performance can be increased. To better understand the morphological basis of high performance and thermal robustness of elastically powered movements, we compared feeding dynamics at a range of temperatures (5-25°C) in two species of terrestrial plethodontid salamanders, Plethodon metcalfi and Ensatina eschscholtzii, which differ in tongue muscle architecture and the mechanism of tongue projection. We found that Ensatina is capable of ballistic projection with a mean muscle mass-specific power of 2100 W kg(-1), revealing an elastic mechanism. Plethodon, in contrast, projected its tongue non-ballistically with a mean power of only 18 W kg(-1), indicating it is muscle powered. Ensatina projected its tongue significantly farther than Plethodon and with dynamics that had significantly lower thermal sensitivity at temperatures below 15°C. These performance differences were correlated with morphological differences, namely elongated collagenous aponeuroses in the projector muscle of Ensatina as compared with Plethodon, which are likely the site of energy storage, and the absence in Ensatina of projector muscle fibers attaching to the tongue skeleton that allows projection to be truly ballistic. These findings demonstrate that, in these otherwise similar species, the presence in one species of elaborated connective tissue in series with myofibers confers not only 10-fold greater absolute performance but also greater thermal robustness of performance. We conclude that changes in muscle and connective tissue architecture are sufficient to alter significantly the mechanics, performance and thermal robustness of musculoskeletal systems.

Running for your life or running for your dinner: what drives fiber-type evolution in lizard locomotor muscles?

Despite its role in whole-animal performance, the adaptation of muscle physiology related to terrestrial locomotion remains underexplored. We tested evolutionary models based on predator escape and foraging strategies of lizards to assess whether fiber-type composition of a leg muscle is adaptive for behavior. The best-fitting model for fast-twitch fiber-type evolution was one based on predator-escape strategy, while the foraging-mode model fared poorly (Akaike Information Criterion with small sample size correction; DeltaAICc=29.7). According to the predator-escape model, lizards relying on sprints to avoid predators are predicted to have relatively higher proportions of fast glycolytic (FG) fibers (70%), while cryptic lizards are predicted to have relatively higher fast oxidative glycolytic (FOG) fiber proportions (77%). This pattern suggests an evolutionary trend toward greater FG (FOG) fiber composition among lizards that specialize in sprinting (crypsis). The best-fitting model for slow-twitch fibers had a single optimum, suggesting a common selective pressure across these lizards. The second-best model explaining slow-twitch fiber-type evolution was Brownian motion (DeltaAICc=0.80), indicating some support for neutral evolution. We find evidence suggesting that different fiber types occurring in the same muscle can evolve under different evolutionary pressures.

Adjusting muscle function to demand: joint work during acceleration in wild turkeys.

We measured the net work performed at hind limb joints in running turkeys to determine the source of mechanical power for acceleration. We tested the hypothesis that net mechanical work per step increases in proportion to acceleration at all four major hind limb joints (hip, knee, ankle and tarsometatarsal-phalangeal joint). This hypothesis was based on the idea that all hind limb muscles should contribute mechanical work to maximize performance during accelerations, and a previous study that indicated the mechanical power output of the entire turkey hind limb musculature was remarkably high. We used high-speed video and force-plate measurements to measure joint moment, velocity and power output during single foot-contacts of running accelerations. By measuring steps in which the animals were relatively more or less motivated to accelerate, we obtained data for a range of accelerations, all at approximately the same running speed. Net joint work per step increased at the hip and ankle as a function of acceleration. Hip net work per unit body mass was 0.12+/-0.09 J kg(-1) averaged over the five lowest accelerations (-0.22+/-0.08 m s(-2)), and 0.87+/-0.20 J kg(-1) for the five highest accelerations (4.86+/-0.27 m s(-2)). Ankle work was -0.21+/-0.11 J kg(-1) for the lowest accelerations and 0.71+/-0.28 J kg(-1) for the highest. The high work output at the ankle is consistent with the idea that elastic mechanisms function to increase muscle work during acceleration. The work performed at the knee and tarsometatarsal-phalangeal joint was independent of acceleration in a step. These results support the idea that hip and ankle extensors contribute significantly to the work necessary to accelerate the body. We also measured the change in joint moment and angular excursion with acceleration to determine whether the mechanism for increasing work output at a joint involved an increase in muscle force or muscle shortening. The increase in joint work at the hip and ankle resulted almost entirely from an increase in joint angular excursion during stance. Hip extension increased by more than threefold from the lowest to the highest accelerations, and the angular excursion of the ankle increased from -24.8+/-4.7 degrees (net flexion) at the lowest accelerations to 33.0+/-12.8 degrees (net extension) at the highest accelerations. Mean stance joint moment was unchanged with acceleration at the ankle and increased by approximately 35% at the hip across the range of accelerations. These patterns of joint moment and excursion indicate that turkeys increase mechanical work for acceleration primarily by increasing muscle shortening, rather than muscle force.

Mechanical power output during running accelerations in wild turkeys.

We tested the hypothesis that the hindlimb muscles of wild turkeys (Meleagris gallopavo) can produce maximal power during running accelerations. The mechanical power developed during single running steps was calculated from force-plate and high-speed video measurements as turkeys accelerated over a trackway. Steady-speed running steps and accelerations were compared to determine how turkeys alter their running mechanics from a low-power to a high-power gait. During maximal accelerations, turkeys eliminated two features of running mechanics that are characteristic of steady-speed running: (i) they produced purely propulsive horizontal ground reaction forces, with no braking forces, and (ii) they produced purely positive work during stance, with no decrease in the mechanical energy of the body during the step. The braking and propulsive forces ordinarily developed during steady-speed running are important for balance because they align the ground reaction force vector with the center of mass. Increases in acceleration in turkeys correlated with decreases in the angle of limb protraction at toe-down and increases in the angle of limb retraction at toe-off. These kinematic changes allow turkeys to maintain the alignment of the center of mass and ground reaction force vector during accelerations when large propulsive forces result in a forward-directed ground reaction force. During the highest accelerations, turkeys produced exclusively positive mechanical power. The measured power output during acceleration divided by the total hindlimb muscle mass yielded estimates of peak instantaneous power output in excess of 400 W kg(-1) hindlimb muscle mass. This value exceeds estimates of peak instantaneous power output of turkey muscle fibers. The mean power developed during the entire stance phase increased from approximately zero during steady-speed runs to more than 150 W kg(-1) muscle during the highest accelerations. The high power outputs observed during accelerations suggest that elastic energy storage and recovery may redistribute muscle power during acceleration. Elastic mechanisms may expand the functional range of muscle contractile elements in running animals by allowing muscles to vary their mechanical function from force-producing struts during steady-speed running to power-producing motors during acceleration.