Publications

2020
Ross SA, Rimkus B, Konow N, Biewener AA, Wakeling JM. Added mass in rat plantaris muscle causes a reduction in mechanical work. Journal of Experimental Biology [Internet]. 2020;223 (19). Publisher's VersionAbstract
Most of what we know about whole muscle behaviour comes from experiments on single fibres or small muscles that are scaled up in size without considering the effects of the additional muscle mass. Previous modelling studies have shown that tissue inertia acts to slow the rate of force development and maximum velocity of muscle during shortening contractions and decreases the work and power per cycle during cyclic contractions; however, these results have not yet been confirmed by experiments on living tissue. Therefore, in this study we conducted in situ work-loop experiments on rat plantaris muscle to determine the effects of increasing the mass of muscle on mechanical work during cyclic contractions. We additionally simulated these experimental contractions using a mass-enhanced Hill-type model to validate our previous modelling work. We found that greater added mass resulted in lower mechanical work per cycle relative to the unloaded trials in which no mass was added to the muscle (P=0.041 for both 85 and 123% increases in muscle mass). We additionally found that greater strain resulted in lower work per cycle relative to unloaded trials at the same strain to control for length change and velocity effects on the work output, possibly due to greater accelerations of the muscle mass at higher strains. These results confirm that tissue mass reduces muscle mechanical work at larger muscle sizes, and that this effect is likely amplified for lower activations.
Taylor-Burt KR, Biewener AA. Aquatic and terrestrial takeoffs require different hindlimb kinematics and muscle function in mallard ducks. Journal of Experimental Biology [Internet]. 2020;223 (16). Publisher's VersionAbstract

Mallard ducks are capable of performing a wide range of behaviors including nearly vertical takeoffs from both terrestrial and aquatic habitats. The hindlimb plays a key role during takeoffs from both media. However, because force generation differs in water versus on land, hindlimb kinematics and muscle function are likely modulated between these environments. Specifically, we hypothesize that hindlimb joint motion and muscle shortening are faster during aquatic takeoffs, but greater hindlimb muscle forces are generated during terrestrial takeoffs. In this study, we examined the hindlimb kinematics and in vivo contractile function of the lateral gastrocnemius (LG), a major ankle extensor and knee flexor, during takeoffs from water versus land in mallard ducks. In contrast to our hypothesis, we observed no change in ankle angular velocity between media. However, the hip and metatarsophalangeal joints underwent large excursions during terrestrial takeoffs but exhibited almost no motion during aquatic takeoffs. The knee extended during terrestrial takeoffs but flexed during aquatic takeoffs. Correspondingly, LG fascicle shortening strain, shortening velocity and pennation angle change were greater during aquatic takeoffs than during terrestrial takeoffs because of the differences in knee motion. Nevertheless, we observed no significant differences in LG stress or work, but did see an increase in muscle power output during aquatic takeoffs. Because differences in the physical properties of aquatic and terrestrial media require differing hindlimb kinematics and muscle function, animals such as mallards may be challenged to tune their muscle properties for movement across differing environments.

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Veiga GN, Biewener AA, Fuller A, van de Ven TMFN, McGowan CP, Panaino W, Snelling EP. Functional morphology of the ankle extensor muscle-tendon units in the springhare Pedetes capensis shows convergent evolution with macropods for bipedal hopping locomotion. Journal of Anatomy. 2020;237 (3) :568-578.Abstract
This study assesses the functional morphology of the ankle extensor muscle-tendon units of the springhare Pedetes capensis, an African bipedal hopping rodent, to test for convergent evolution with the Australian bipedal hopping macropods. We dissect and measure the gastrocnemius, soleus, plantaris, and flexor digitorum longus in 10 adult springhares and compare them against similar-sized macropods using phylogenetically informed scaling analyses. We show that springhares align reasonably well with macropod predictions, being statistically indistinguishable with respect to the ankle extensor mean weighted muscle moment arm (1.63 vs. 1.65 cm, respectively), total muscle mass (41.1 vs. 29.2 g), total muscle physiological cross-sectional area (22.9 vs. 19.3 cm2), mean peak tendon stress (26.2 vs. 35.2 MPa), mean tendon safety factor (4.7 vs. 3.6), and total tendon strain energy return capacity (1.81 vs. 1.82 J). However, total tendon cross-sectional area is significantly larger in springhares than predicted for a similar-sized macropod (0.26 vs. 0.17 cm2, respectively), primarily due to a greater plantaris tendon thickness (0.084 vs. 0.048 cm2), and secondarily because the soleus muscle-tendon unit is present in springhares but is vestigial in macropods. The overall similarities between springhares and macropods indicate that evolution has favored comparable lower hindlimb body plans for bipedal hopping locomotion in the two groups of mammals that last shared a common ancestor ~160 million years ago. The springhare’s relatively thick plantaris tendon may facilitate rapid transfer of force from muscle to skeleton, enabling fast and accelerative hopping, which could help to outpace and outmaneuver predators.
Gordon JC, Holt NC, Biewener AA, Daley MA. Tuning of feedforward control enables stable muscle force-length dynamics after loss of autogenic proprioceptive feedback. eLife [Internet]. 2020. Publisher's VersionAbstract
Animals must integrate feedforward, feedback and intrinsic mechanical control mechanisms to maintain stable locomotion. Recent studies of guinea fowl (Numida meleagris) revealed that the distal leg muscles rapidly modulate force and work output to minimize perturbations in uneven terrain. Here we probe the role of reflexes in the rapid perturbation responses of muscle by studying the effects of proprioceptive loss. We induced bilateral loss of autogenic proprioception in the lateral gastrocnemius muscle (LG) using self-reinnervation. We compared in vivo muscle dynamics and ankle kinematics in birds with reinnervated and intact LG. Reinnervated and intact LG exhibit similar steady state mechanical function and similar work modulation in response to obstacle encounters. Reinnervated LG exhibits 23ms earlier steady-state activation, consistent with feedforward tuning of activation phase to compensate for lost proprioception. Modulation of activity duration is impaired in rLG, confirming the role of reflex feedback in regulating force duration in intact muscle.
Konow N, Collias A, Biewener AA. Skeletal Muscle Shape Change in Relation to Varying Force Requirements Across Locomotor Conditions. Frontiers in Physiology [Internet]. 2020;11 (143). Publisher's VersionAbstract
Contractions of skeletal muscles to generate in vivo movement involve dynamic changes in contractile and elastic tissue strains that likely interact to influence the force and work of a muscle. However, studies of the in vivo dynamics of skeletal muscle and tendon strains remain largely limited to bipedal animals, and rarely cover the broad spectra of movement requirements met by muscles that operate as motors, struts, or brakes across the various gaits that animals commonly use and conditions they encounter. Using high-speed bi-planar fluoromicrometry, we analyze in vivo strains within the rat medial gastrocnemius (MG) across a range of gait and slope conditions. These conditions require changes in muscle force ranging from decline walk (low) to incline gallop (high). Measurements are made from implanted (0.5–0.8 mm) tantalum spheres marking MG mid-belly width, mid-belly thickness, as well as strains of distal fascicles, the muscle belly, and the Achilles tendon. During stance, as the muscle contracts, muscle force increases linearly with respect to gait–slope combinations, and both shortening and lengthening fiber strains increase from approximately 5 to 15% resting length. Contractile change in muscle thickness (thickness strain) decreases (r2 = 0.86; p = 0.001); whereas, the change in muscle width (width strain) increases (r2 = 0.88; p = 0.001) and tendon strain increases (r2 = 0.77; p = 0.015). Our results demonstrate force-dependency of contractile and tendinous tissue strains with compensatory changes in shape for a key locomotor muscle in the hind limb of a small quadruped. These dynamic changes are linked to the ability of a muscle to tune its force and work output as requirements change with locomotor speed and environmental conditions.
Rull M, Solomon J, Konow N. Elastic recoil action amplifies jaw closure in an aquatic-feeding salamander. Proceedings of the Royal Society Ser. B [Internet]. 2020;287 (1927). Publisher's Version
Taylor-Burt K, Konow N, Biewener AA. Post-activation muscle potentiation and its relevance to cyclical behaviours. Biol. Letters [Internet]. 2020;16 (6) :1-6. Publisher's VersionAbstract
Muscle can experience post-activation potentiation (PAP), a temporary increase in force and rate of force development, when contractions are closely timed; therefore, cyclical behaviours are likely affected by PAP, as succeeding contraction cycles can lead to potentiation over several subsequent cycles. Here, we examined PAP during in situ cyclical contractions of the mallard lateral gastrocnemius (LG). Surface swimming, a cyclical behaviour, was mimicked with work-loops using in vivo LG length change and stimulation parameters. Tests were performed at mallards' preferred cycle frequency as well as at lower and higher frequencies. Like muscles from mammals, anurans and arthropods, the mallard LG exhibited PAP with increases in peak force, average force rate and net work. Staircase potentiation occurred over two or more work-loop cycles, resulting in gradual increases in PAP. The number of cycles needed to reach maximum work varied with cycle frequency, requiring more cycles at higher cycle frequencies. PAP occurred under in vivo-like stimulation parameters, suggesting a potentially important role of PAP in animal locomotion, especially in cyclical behaviours.
Jayshini NM, Kessler S, Rainbow MJ, D'Andrea SE, Konow N, Kelly L, Lichtwark GA. The Reliability of Foot and Ankle Bone and Joint Kinematics Measured With Biplanar Videoradiography and Manual Scientific Rotoscoping. Frontiers in Bioengineering and Biotechnology [Internet]. 2020. Publisher's VersionAbstract
The intricate motion of the small bones of the feet are critical for its diverse function. Accurately measuring the 3-dimensional (3D) motion of these bones has attracted much attention over the years and until recently, was limited to invasive techniques or quantification of functional segments using multi-segment foot models. Biplanar videoradiography and model-based scientific rotoscoping offers an exciting alternative that allows us to focus on the intricate motion of individual bones in the foot. However, scientific rotoscoping, the process of rotating and translating a 3D bone model so that it aligns with the captured x-ray images, is either semi- or completely manual and it is unknown how much human error affects tracking results. Thus, the aim of this study was to quantify the inter- and intra-operator reliability of manually rotoscoping in vivo bone motion of the tibia, talus, and calcaneus during running. Three-dimensional CT bone volumes and high-speed biplanar videoradiography images of the foot were acquired on six participants. The six-degree-of-freedom motions of the tibia, talus, and calcaneus were determined using a manual markerless registration algorithm. Two operators performed the tracking, and additionally, the first operator re-tracked all bones, to test for intra-operator effects. Mean RMS errors were 1.86 mm and 1.90° for intra-operator comparisons and 2.30 mm and 2.60° for inter-operator comparisons across all bones and planes. The moderate to strong similarity values indicate that tracking bones and joint kinematics between sessions and operators is reliable for running. These errors are likely acceptable for defining gross joint angles. However, this magnitude of error may limit the capacity to perform advanced analyses of joint interactions, particularly those that require precise (sub-millimeter) estimates of bone position and orientation. Optimizing the view and image quality of the biplanar videoradiography system as well as the automated tracking algorithms for rotoscoping bones in the foot are required to reduce these errors and the time burden associated with the manual processing.
Schwarz D, Konow N, Roba YT, Heiss E. A salamander that chews with complex, three-dimensional mandible movements . Journal of Experimental Biology [Internet]. 2020. Publisher's VersionAbstract
It is generally accepted that most non-mammal tetrapods have a hinge-like jaw operation restricted to vertical opening and closing movements. Many mammal jaw joints, by contrast, operate in more complex, three-dimensional ways, involving not only vertical, but also propalinal (rostro-caudal) and transverse (lateral) movements. Data on intraoral food processing in lissamphibians and sauropsids has prompted a generally accepted view that these groups mostly swallow food unreduced, and that in those cases where lissamphibians and sauropsids chew, they mostly use simple vertical jaw movements for food processing. The exception to that generally accepted view being some propalinal chewing in sauropsids. We combined 3D kinematics and morphological analyses from biplanar high-speed video fluoroscopy and micro-CT to determine how the paedomorphic salamander Siren intermedia treats captured food. We discovered that S. intermedia not only uses intraoral food processing, but that the elaborated morphology of its jaw joint facilitates mandibular motions in all three planes, resulting in complex three-dimensional chewing. Thus, our data challenge the commonly held view that complex three-dimensional chewing movements are exclusive to mammals, by suggesting that complex chewing mechanisms might evolved early in tetrapod evolution.
Ravi S, Noda R, Gagliardi S, Kolomenskiy D, Combes S, Liu H, Biewener A, Konow N. Modulation of Flight Muscle Recruitment and Wing Rotation Enables Hummingbirds to Mitigate Aerial Roll Perturbations. Current Biology [Internet]. 2020;30 (2) :187-195.e4. Publisher's VersionAbstract
Both biological and artificial fliers must contend with aerial perturbations that are ubiquitous in the outdoor environment. Flapping fliers are generally least stable, but also the most maneuverable in roll, yet roll control in biological fliers remains less well understood. Hummingbirds are suitable models for linking aerodynamic perturbations to flight control strategies, as these small, powerful fliers are capable of remaining airborne even in adverse airflows. We challenged hummingbirds to fly within a longitudinally oriented vortex that imposed a continuous roll perturbation, measured wing kinematics and neuromotor activation of the major flight muscles with synchronized high-speed video and electromyography and used computational fluid dynamics (CFD) to estimate the aerodynamic forces generated by wing motions. Hummingbirds responded to the perturbation using bilaterally different activation of the main flight muscles and maintained symmetry in most major aspects of wing motion including stroke amplitude, stroke plane angle, and flapping frequency. However, hummingbirds also displayed consistent bilateral differences in subtle wing kinematics traits, including wing rotation and elevation. CFD modeling implicate asymmetric responses in wing rotation as important for generating the necessary stabilizing torques, suggesting that intrinsic wing muscles play a critical role in aerodynamic control. The birds also augment flight stabilization by adjusting body and tail posture to expose greater surface area to upwash than to the undesirable downwash. Our results provide insight into the remarkable capacity of hummingbirds to maintain flight control and bio-inspiration for simple yet effective control strategies for robotic fliers to contend with unfamiliar and challenging real-word aerial conditions.
Fahn-Lai P, Biewener AA, Pierce SE. Broad similarities in shoulder muscle architecture and organization across two amniotes: implications for reconstructing non-mammalian synapsids. PeerJ [Internet]. 2020;8 (e8556). Publisher's VersionAbstract
The evolution of upright limb posture in mammals may have enabled modifications of the forelimb for diverse locomotor ecologies. A rich fossil record of non-mammalian synapsids holds the key to unraveling the transition from “sprawling” to “erect” limb function in the precursors to mammals, but a detailed understanding of muscle functional anatomy is a necessary prerequisite to reconstructing postural evolution in fossils. Here we characterize the gross morphology and internal architecture of muscles crossing the shoulder joint in two morphologically-conservative extant amniotes that form a phylogenetic and morpho-functional bracket for non-mammalian synapsids: the Argentine black and white tegu Salvator merianae and the Virginia opossum Didelphis virginiana. By combining traditional physical dissection of cadavers with nondestructive three-dimensional digital dissection, we find striking similarities in muscle organization and architectural parameters. Despite the wide phylogenetic gap between our study species, distal muscle attachments are notably similar, while differences in proximal muscle attachments are driven by modifications to the skeletal anatomy of the pectoral girdle that are well-documented in transitional synapsid fossils. Further, correlates for force production, physiological cross-sectional area (PCSA), muscle gearing (pennation), and working range (fascicle length) are statistically indistinguishable for an unexpected number of muscles. Functional tradeoffs between force production and working range reveal muscle specializations that may facilitate increased girdle mobility, weight support, and active stabilization of the shoulder in the opossum—a possible signal of postural transformation. Together, these results create a foundation for reconstructing the musculoskeletal anatomy of the non-mammalian synapsid pectoral girdle with greater confidence, as we demonstrate by inferring shoulder muscle PCSAs in the fossil non-mammalian cynodont Massetognathus pascua
2019
Clites TR, Arnold AS, Singh NM, Kline E, H.Chen, Tugman C, Billadeau B, Biewener AA, Herr HM. Goats Decrease Hindlimb Stiffness When Walking Over Compliant Surfaces. Journal of Experimental Biology. 2019.Abstract
Leg stiffness, commonly estimated as the 'compression' of a defined leg element in response to a load, has long been used to characterize terrestrial locomotion. This study investigated how goats adjust the stiffness of their hindlimbs to accommodate surfaces of different stiffness. Goats provide a compelling animal model for studying leg stiffness modulation, because they skillfully ambulate over a range of substrates that vary in compliance. To investigate the adjustments that goats make when walking over such substrates, ground reaction forces and three-dimensional trajectories of hindlimb markers were recorded as goats walked on rigid, rubber and foam surfaces. Net joint moments, power and work at the hip, knee, ankle and metatarsophalangeal joints were estimated throughout stance via inverse dynamics. Hindlimb stiffness was estimated from plots of total leg force versus total leg length, and individual joint stiffness was estimated from plots of joint moment versus joint angle. Our results support the hypothesis that goats modulate hindlimb stiffness in response to surface stiffness; specifically, hindlimb stiffness decreased on the more compliant surfaces (P<0.002). Estimates of joint stiffness identified hip and ankle muscles as the primary drivers of these adjustments. When humans run on compliant surfaces, they generally increase leg stiffness to preserve their center-of-mass mechanics. We did not estimate center-of-mass mechanics in this study; nevertheless, our estimates of hindlimb stiffness suggest that goats exhibit a different behavior. This study offers new insight into mechanisms that allow quadrupeds to modulate their gait mechanics when walking on surfaces of variable compliance.
Eng CM, Konow N, Tijs C, Holt NC, Biewener AA. In vivo force–length and activation dynamics of two distal rat hindlimb muscles in relation to gait and grade. Journal of Experimental Biology [Internet]. 2019. Publisher's VersionAbstract
Muscle function changes to meet the varying mechanical demands of locomotion across different gait and grade conditions. A muscle's work output is determined by time-varying patterns of neuromuscular activation, muscle force and muscle length change, but how these patterns change under different conditions in small animals is not well defined. Here, we report the first integrated in vivo force–length and activation patterns in rats, a commonly used small animal model, to evaluate the dynamics of two distal hindlimb muscles (medial gastrocnemius and plantaris) across a range of gait (walk, trot and gallop) and grade (level and incline) conditions. We use these data to explore how the pattern of force production, muscle activation and muscle length changes across conditions in a small quadrupedal mammal. As hypothesized, we found that the rat muscles show limited fascicle strains during active force generation in stance across gaits and grades, indicating that these distal rat muscles generate force economically but perform little work, similar to patterns observed in larger animals during level locomotion. Additionally, given differences in fiber type composition and variation in motor unit recruitment across the gait and grade conditions examined here for these muscles, the in vivo force–length behavior and neuromuscular activation data reported here can be used to validate improved two-element Hill-type muscle models.
Lai A, Biewener AA, Wakeling JM. Muscle-specific indices to characterise the functional behavior of human lower-limb muscles during locomotion . Journal of Biomechanics [Internet]. 2019;89 :134-138. Publisher's Version
McHorse BK, Biewener AA, Pierce SE. The evolution of a single toe in horses: causes, consequences, and the way forward. Integrative & Compartive Biology [Internet]. 2019;icz050 (10.1093) :1-18. Publisher's Version
Kessler SE, Rainbow MJ, Lichtwark GA, Cresswell AG, D'Andrea SE, Konow N, Kelly LA. A Direct Comparison of Biplanar Videoradiography and Optical Motion Capture for Foot and Ankle Kinematics. Frontiers in Bioengineering and Biotechnology [Internet]. 2019;7 (199) :1-10. Publisher's VersionAbstract
Measuring motion of the human foot presents a unique challenge due to the large number of closely packed bones with congruent articulating surfaces. Optical motion capture (OMC) and multi-segment models can be used to infer foot motion, but might be affected by soft tissue artifact (STA). Biplanar videoradiography (BVR) is a relatively new tool that allows direct, non-invasive measurement of bone motion using high-speed, dynamic x-ray images to track individual bones. It is unknown whether OMC and BVR can be used interchangeably to analyse multi-segment foot motion. Therefore, the aim of this study was to determine the agreement in kinematic measures of dynamic activities. Nine healthy participants performed three walking and three running trials while BVR was recorded with synchronous OMC. Bone position and orientation was determined through manual scientific-rotoscoping. The OMC and BVR kinematics were co-registered to the same coordinate system, and BVR tracking was used to create virtual markers for comparison to OMC during dynamic trials. Root mean square (RMS) differences in marker positions and joint angles as well as a linear fit method (LFM) was used to compare the outputs of both methods. When comparing BVR and OMC, sagittal plane angles were in good agreement (ankle: R2 = 0.947, 0.939; Medial Longitudinal Arch (MLA) Angle: R2 = 0.713, 0.703, walking and running, respectively). When examining the ankle, there was a moderate agreement between the systems in the frontal plane (R2 = 0.322, 0.452, walking and running, respectively), with a weak to moderate correlation for the transverse plane (R2 = 0.178, 0.326, walking and running, respectively). However, root mean squared error (RMSE) showed angular errors ranging from 1.06 to 8.31° across the planes (frontal: 3.57°, 3.67°, transverse: 4.28°, 4.70°, sagittal: 2.45°, 2.67°, walking and running, respectively). Root mean square (RMS) differences between OMC and BVR marker trajectories were task dependent with the largest differences in the shank (6.0 ± 2.01 mm) for running, and metatarsals (3.97 ± 0.81 mm) for walking. Based on the results, we suggest BVR and OMC provide comparable solutions to foot motion in the sagittal plane, however, interpretations of out-of-plane movement should be made carefully.
Witzmann F, Brainerd EL, Konow N. Eye Movements in Frogs and Salamanders—Testing the Palatal Buccal Pump Hypothesis. Integrative Organismal Biology [Internet]. 2019;1 (1). Publisher's VersionAbstract
In frogs and salamanders, movements of the eyeballs in association with an open palate have often been proposed to play a functional role in lung breathing. In this “palatal buccal pump,” the eyeballs are elevated during the lowering of the buccal floor to suck air in through the nares, and the eyeballs are lowered during elevation of the buccal floor to help press air into the lungs. Here, we used X-Ray Reconstruction of Moving Morphology to investigate eye movements during lung breathing and feeding in bullfrogs and axolotls. Our data do not show eye movements that would be in accordance with the palatal buccal pump. On the contrary, there is a small passive elevation of the eyeballs when the buccal floor is raised. Inward drawing of the eyeballs occurs only during body motion and for prey transport in bullfrogs, but this was not observed in axolotls. Each eye movement in bullfrogs has a vertical, a mediolateral, and an anteroposterior component. Considering the surprisingly weak posterior motion component of the eyeballs, their main role in prey transport might be fixing the prey by pressing it against the buccal floor. The retraction of the buccal floor would then contribute to the posterior push of the prey. Because our study provides no evidence for a palatal buccal pump in frogs and salamanders, there is also no experimental support for the idea of a palatal buccal pump in extinct temnospondyl amphibians, in contrast to earlier suggestions.
Arellano C, Konow N, Gidmark N, Roberts TJ. Evidence of a tunable biological spring: Elastic energy storage in aponeuroses varies with transverse strain in vivo. Proceedings of Royal Society Series B [Internet]. 2019;286 (1900) :20182764. Publisher's Version
Lin Y, Konow N, Dumont ER. How moles destroy your lawn: the forelimb kinematics of Eastern moles in loose and compact substrates. Journal of Experimental Biology [Internet]. 2019;222. Publisher's Version
Heiss E, Schwartz D, Konow N. Chewing or not? Intraoral food processing in a salamandrid newt. Journal of Experimental Biology [Internet]. 2019;222. Publisher's Version

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