Kinematic trajectories in response to speed perturbations in walking suggest modular task-level control of leg angle and length.

Navigating complex terrains requires dynamic interactions between the substrate, musculoskeletal and sensorimotor systems. Current perturbation studies have mostly used visible terrain height perturbations, which do not allow us to distinguish among the neuromechanical contributions of feedforward control, feedback-mediated and mechanical perturbation responses. Here, we use treadmill belt speed perturbations to induce a targeted perturbation to foot speed only, and without terrain-induced changes in joint posture and leg loading at stance onset. Based on previous studies suggesting a proximo-distal gradient in neuromechanical control, we hypothesized that distal joints would exhibit larger changes in joint kinematics, compared to proximal joints. Additionally, we expected birds to use feedforward strategies to increase the intrinsic stability of gait. To test these hypotheses, seven adult guinea fowl were video recorded while walking on a motorized treadmill, during both steady and perturbed trials. Perturbations consisted of repeated exposures to a deceleration and acceleration of the treadmill belt speed. Surprisingly, we found that joint angular trajectories and center of mass fluctuations remain very similar, despite substantial perturbation of foot velocity by the treadmill belt. Hip joint angular trajectories exhibit the largest changes, with the birds adopting a slightly more flexed position across all perturbed strides. Additionally, we observed increased stride duration across all strides, consistent with feedforward changes in the control strategy. The speed perturbations mainly influenced the timing of stance and swing, with the largest kinematic changes in the strides directly following a deceleration. Our findings do not support the general hypothesis of a proximo-distal gradient in joint control, as distal joint kinematics remain largely unchanged. Instead, we find that leg angular trajectory and the timing of stance and swing are most sensitive to this specific perturbation, and leg length actuation remains largely unchanged. Our results are consistent with modular task-level control of leg length and leg angle actuation, with different neuromechanical control and perturbation sensitivity in each actuation mode. Distal joints appear to be sensitive to changes in vertical loading but not foot fore-aft velocity. Future directions should include in vivo studies of muscle activation and force-length dynamics to provide more direct evidence of the sensorimotor control strategies for stability in response to belt speed perturbations.

From Tusko to Titin: the role for comparative physiology in an era of molecular discovery.

As we approach the centenary of the term "comparative physiology," we reexamine its role in modern biology. Finding inspiration in Krogh's classic 1929 paper, we first look back to some timeless contributions to the field. The obvious and fascinating variation among animals is much more evident than is their shared physiological unity, which transcends both body size and specific adaptations. The "unity in diversity" reveals general patterns and principles of physiology that are invisible when examining only one species. Next, we examine selected contemporary contributions to comparative physiology, which provides the context in which reductionist experiments are best interpreted. We discuss the sometimes surprising insights provided by two comparative "athletes" (pronghorn and rattlesnakes), which demonstrate 1) animals are not isolated molecular mechanisms but highly integrated physiological machines, a single "rate-limiting" step may be exceptional; and 2) extremes in nature are rarely the result of novel mechanisms, but rather employ existing solutions in novel ways. Furthermore, rattlesnake tailshaker muscle effectively abolished the conventional view of incompatibility of simultaneous sustained anaerobic glycolysis and oxidative ATP production. We end this review by looking forward, much as Krogh did, to suggest that a comparative approach may best lend insights in unraveling how skeletal muscle stores and recovers mechanical energy when operating cyclically. We discuss and speculate on the role of the largest known protein, titin (the third muscle filament), as a dynamic spring capable of storing and recovering elastic recoil potential energy in skeletal muscle.

Functional implications of supercontracting muscle in the chameleon tongue retractors.

Chameleons capture prey items using a ballistic tongue projection mechanism that is unique among lizards. During prey capture, the tongue can be projected up to two full body lengths and may extend up to 600 % of its resting length. Being ambush predators, chameleons eat infrequently and take relatively large prey. The extreme tongue elongation (sixfold) and the need to be able to retract fairly heavy prey at any given distance from the mouth are likely to place constraints on the tongue retractor muscles. The data examined here show that in vivo retractor force production is almost constant for a wide range of projection distances. An examination of muscle physiology and of the ultrastructure of the tongue retractor muscle shows that this is the result (i) of active hyoid retraction, (ii) of large muscle filament overlap at maximal tongue extension and (iii) of the supercontractile properties of the tongue retractor muscles. We suggest that the chameleon tongue retractor muscles may have evolved supercontractile properties to enable a substantial force to be produced over a wide range of tongue projection distances. This enables chameleons successfully to retract even large prey from a variety of distances in their complex three-dimensional habitat.

Morphology and histochemistry of the hyolingual apparatus in chameleons.

We reexamined the morphological and functional properties of the hyoid, the tongue pad, and hyolingual musculature in chameleons. Dissections and histological sections indicated the presence of five distinctly individualized pairs of intrinsic tongue muscles. An analysis of the histochemical properties of the system revealed only two fiber types in the hyolingual muscles: fast glycolytic and fast oxidative glycolytic fibers. In accordance with this observation, motor-endplate staining showed that all endplates are of the en-plaque type. All muscles show relatively short fibers and large numbers of motor endplates, indicating a large potential for fine muscular control. The connective tissue sheet surrounding the entoglossal process contains elastin fibers at its periphery, allowing for elastic recoil of the hyolingual system after prey capture. The connective tissue sheets surrounding the m. accelerator and m. hyoglossus were examined under polarized light. The collagen fibers in the accelerator epimysium are configured in a crossed helical array that will facilitate limited muscle elongation. The microstructure of the tongue pad as revealed by SEM showed decreased adhesive properties, indicating a change in the prey prehension mechanics in chameleons compared to agamid or iguanid lizards. These findings provide the basis for further experimental analysis of the hyolingual system.

The mechanics of prey prehension in chameleons.

Iguanian lizards generally use their tongue to capture prey. Because lingual prehension is based on surface phenomena (wet adhesion, interlocking), the maximal prey size that can be captured is small. However, published records show that prey items eaten by chameleons include small vertebrates such as lizards and birds, indicating that these lizards are using a different prey prehension mechanism. Using high-speed video recordings, cineradiography, electromyography, nerve transection and stimulation experiments, we investigated the function of the tongue during prey capture. The results of these experiments indicate that chameleons have modified the primitive iguanian system by including a suction component in their prehension mechanism. Suction is generated by the activity of two modified intrinsic tongue muscles that pull the tongue pad inwards. Moreover, we demonstrate that the mechanism described here is a prerequisite for successful feeding.

Comparative study of tongue protrusion in three iguanian lizards, Sceloporus undulatus, Pseudotrapelus sinaitus and Chamaeleo jacksonii.

The goal of this study was to investigate the function of the hyolingual muscles used during tongue protraction in iguanian lizards. High-speed videography and nerve-transection techniques were used to study prey capture in the iguanid Sceloporus undulatus, the agamid Pseudoptrapelus sinaitus and the chameleonid Chamaeleo jacksonii. Denervation of the mandibulohyoideus muscle slips had an effect only on P. sinaitus and C. jacksonii, in which tongue protrusion or projection distance was reduced. In C. jacksonii, denervation of the M. mandibulohyoideus completely prevented little hyoid protraction. Denervation of the M. verticalis had no effect on S. undulatus, but reduced tongue protrusion distance in P. sinaitus. Denervation of the accelerator muscle in C. jacksonii inhibited tongue projection completely. The function of the M. mandibulohyoideus and M. verticalis has become increasingly specialized in P. sinaitus and especially in C. jacksonii to allow greater tongue protrusion. The combined results of these treatments suggest that these three groups represent transitional forms, both morphologically and functionally, in the development of a projectile tongue.

Comparison of isometric contractile properties of the tongue muscles in three species of frogs, Litoria caerulea, Dyscophus guinetti, and Bufo marinus.

Previous studies show that anurans feed in at least three different ways. Basal frogs have a broad tongue that shortens during protraction and emerges only a short distance from the mouth. Some frogs have long, narrow tongues that elongate dramatically due primarily to inertia from mouth opening, which is transferred to the tongue. A few species have a hydrostatic mechanism that produces tongue elongation during protraction. This functional diversity occurs among frogs that share the same two pairs of tongue muscles. Our study compares the isometric contractile properties of these tongue muscles among three frog species that represent each feeding mechanism. Nerves to the paired protractors and retractors were stimulated electrically in each species to record the force properties, contraction speeds, and fatigabilites of these muscles. Few differences were found in the isometric contractile properties of tongue muscles, and the greatest differences were found in the retractors, not the protractors. We propose that the unique arrangement of the tongue muscles in frogs results in a retractor that may also be coactivated with the protractor in order to produce normal tongue protraction. Inertial effects from body, head, and jaw movements, along with clear differences that we found in passive resistance of the tongues to elongation, may explain much of the behavioral variation in tongue use among species.

Neuromuscular control of prey capture in frogs.

While retaining a feeding apparatus that is surprisingly conservative morphologically, frogs as a group exhibit great variability in the biomechanics of tongue protraction during prey capture, which in turn is related to differences in neuromuscular control. In this paper, I address the following three questions. (1) How do frog tongues differ biomechanically? (2) What anatomical and physiological differences are responsible? (3) How is biomechanics related to mechanisms of neuromuscular control? Frog species use three non-exclusive mechanisms to protract their tongues during feeding: (i) mechanical pulling, in which the tongue shortens as its muscles contract during protraction; (ii) inertial elongation, in which the tongue lengthens under inertial and muscular loading; and (iii) hydrostatic elongation, in which the tongue lengthens under constraints imposed by the constant volume of a muscular hydrostat. Major differences among these functional types include (i) the amount and orientation of collagen fibres associated with the tongue muscles and the mechanical properties that this connective tissue confers to the tongue as a whole; and (ii) the transfer of intertia from the opening jaws to the tongue, which probably involves a catch mechanism that increases the acceleration achieved during mouth opening. The mechanisms of tongue protraction differ in the types of neural mechanisms that are used to control tongue movements, particularly in the relative importance of feed-forward versus feedback control, in requirements for precise interjoint coordination, in the size and number of motor units, and in the afferent pathways that are involved in coordinating tongue and jaw movements. Evolution of biomechanics and neuromuscular control of frog tongues provides an example in which neuromuscular control is finely tuned to the biomechanical constraints and opportunities provided by differences in morphological design among species.

Do arm postures vary with the speed of reaching?

Do arm postures vary with the speed of reaching? For reaching movements in one plane, the hand has been observed to follow a similar path regardless of speed. Recent work on the control of more complex reaching movements raises the question of whether a similar "speed invariance" also holds for the additional degrees of freedom. Therefore we examined human arm movements involving initial and final hand locations distributed throughout the three-dimensional (3D) workspace of the arm. Despite this added complexity, arm kinematics (summarized by the spatial orientation of the "plane of the arm" and the 3D curvature of the hand path) changed very little for movements performed over a wide range of speeds. If the total force (dynamic + quasistatic) had been optimized by the control system (e.g., as in a minimization of the change in joint torques or the change in muscular forces), the optimal solution would change with speed; slow movements would reflect the minimal antigravity torques, whereas fast movements would be more strongly influenced by dynamic factors. The speed-invariant postures observed in this study are instead consistent with a hypothesized optimization of only the dynamic forces.

Morphology and mechanics of tongue movement in the African pig-nosed frog Hemisus marmoratum: a muscular hydrostatic model.

The goal of this study was to investigate morphological adaptations associated with hydrostatic elongation of the tongue during feeding in the African pig-nosed frog Hemisus marmoratum. Whereas previous studies had suggested that the tongue of H. marmoratum elongates hydraulically, the anatomical observations reported here favour a muscular hydrostatic mechanism of tongue elongation. H. marmoratum possesses a previously undescribed compartment of the m. genioglossus (m. genioglossus dorsoventralis), which is intrinsic to the tongue and whose muscle fibres are oriented perpendicular to the long axis of the tongue. On the basis of the arrangement and orientation of muscle fibres in the m. genioglossus and m. hyoglossus, we propose a muscular hydrostatic model of tongue movement in which contraction of the m. genioglossus dorsoventralis, together with unfolding of the intrinsic musculature of the tongue, results in a doubling in tongue length. Electron micrographs of sarcomeres from resting and elongated tongues show that no special adaptations of the sarcomeres are necessary to accommodate the observed doubling in tongue length during feeding. Rather, the sarcomeres of the m. genioglossus longitudinalis are strikingly similar to those of anuran limb muscles. The ability to elongate the tongue hydrostatically, conferred by the presence of the m. genioglossus dorsoventralis, is associated with the appearance of several novel aspects of feeding behaviour in H. marmoratum. These include the ability to protract the tongue slowly, thereby increasing capture success, and the ability to aim the tongue in azimuth and elevation relative to the head. Compared with other frogs, the muscular hydrostatic system of H. marmoratum allows more precise, localized and diverse tongue movements. This may explain why the m. genioglossus of H. marmoratum is composed of a larger number of motor units than that of other frogs.

Distribution of hypoglossal motor neurons innervating the prehensile tongue of the African pig-nosed frog, Hemisus marmoratum.

Using retrograde neuronal tracers, a study of the distribution of hypoglossal motor neurons innervating the tongue musculature was performed in the African pig-nosed frog, Hemisus marmoratum. This species is a radically divergent anuran amphibian with a prehensile tongue that can be aimed in three dimensions relative to the head. The results illustrate a unique rostrocaudal distribution of the ventrolateral hypoglossal nucleus and an unusually large number of motor neurons within this cell group. During the evolution of the long, prehensile tongue of Hemisus, the motor neurons innervating the tongue have greatly increased in number and have become more caudally distributed in the brainstem and spinal cord compared to other anurans. These observations have implications for understanding neuronal reconfiguring of motoneurons for novel morphologies requiring new muscle activation patterns.

The functional anatomy and evolution of hypoglossal afferents in the leopard frog, Rana pipiens.

Previously, we suggested that afferents are present in the hypoglossal nerve of the leopard frog, Rana pipiens. The basis for this was behavioral data obtained after transection of the hypoglossal nerve. These afferents coordinate the timing of tongue protraction with mouth opening during feeding. The goal of the present study was to define anatomically these hypoglossal afferents in Rana pipiens. Retrograde tracing was performed using horseradish peroxidase, fluorescent dextran amines and neurobiotin. Data show that the cell bodies of hypoglossal afferents are located in the dorsal root ganglion of the third spinal nerve and enter the brainstem through its dorsal root. The afferents ascend in the dorsomedial funiculus and move laterally after they pass the obex. They project in the granular layer of the cerebellum and the medial reticular formation. The cervical afferents that travel in this pathway are known to carry proprioceptive and cutaneous sensory information. We hypothesize that the presence of afferents in the hypoglossal nerve is a derived characteristic of anurans, which has resulted from the re-routing of afferent fibers from the third spinal nerve into the hypoglossal nerve. The appearance of hypoglossal afferents coincides with the morphological acquisition of a highly protrusible tongue.

Genome size, secondary simplification, and the evolution of the brain in salamanders.

Compared to other vertebrates, even including lampreys and hagfishes in some respects, salamanders exhibit a relatively simple organization of brain and sense organs which is illustrated here using the visual system as an example. The greatest simplicity is found in the bolitoglossine salamanders, yet all bolitoglossines possess highly projectile tongues and rely on vision for survival; furthermore, some species are agile and acrobatic. The unusual features of the visual system of salamanders include small numbers of large neurons, a low degree of morphological differentiation among neurons, a small proportion of myelinated axons in the optic nerve, and an optic tectum consisting essentially of a periventricular cellular layer and a superficial fiber layer. Similar features are found throughout the central nervous system of salamanders and in the lateral line, auditory and olfactory systems as well. Phylogenetic analysis shows that the most parsimonious interpretation of these data is that the simple organization of the brain and sense organs of salamanders was derived secondarily from a more complex ancestral state. We hypothesize that increased genome size has led to simplification of the nervous system in salamanders. Increased genome size appears to have had profound effects on neural development in salamanders, leading to paedomorphosis, the retention of juvenile or even embryonic characteristics into adulthood. In particular, large genome size is associated with large cell size and reduced rates of cell proliferation, migration and differentiation. Secondary simplification has constrained the function of the salamanders' visual system, primarily by increasing cell size and decreasing cell numbers. However, it also has provided an opportunity for the evolution of compensating mechanisms, which have helped to restore or even enhance visual function. Most apparent among the compensatory mechanisms of bolitoglossine salamanders is the presence of well developed ipsilateral retinotectal projections, which apparently enhance depth perception. It is difficult to explain the unusual history of the nervous system in salamanders solely in terms of natural selection and adaptation. Increasing genome size through selfish replication appears to have played a major role in the evolution of salamander brains by imposing functional constraints as well as creating opportunities for overcoming them.

Evolution of forelimb movement patterns for prey manipulation in anurans.

Unlike other amphibians, frogs often use their forelimbs to capture and transport prey. In the present study, high-speed videography was used to observe forelimb use during feeding in a diverse group of anurans in order to determine the evolution of forelimb movement patterns among anuran taxa. Data were gathered from 488 individuals representing 104 species, 55 genera, and 16 families. Five distinct behavior patterns were identified: scooping entails using the back of the hand to push prey into the mouth; wiping involves the use of the palm of the hand to push prey, protruding laterally from the mouth, toward the midline; during prey stretching, one end of the prey is held in a stationary position by the hands while the other end is pulled upward by the jaws; in grasping, the palms face the midline or the substrate as the fingers are wrapped around the prey; grasping with wrist rotation is similar to grasping, but the wrists rotate inward as the hands grasp the prey so that the palms face the mouth. The distribution of these behavior patterns was mapped onto the most recent phylogenetic hypothesis for anurans. Maximum parsimony analyses suggest that scooping and wiping are primitive and have been retained by many frog lineages. Wiping was not observed in the pipids, which are the only anurans that lack tongues and use hydraulic transport. Prey stretching appears to have evolved several times in unrelated taxa. Grasping and grasping with wrist rotation appear to have evolved only in arboreal groups, suggesting that the ability to climb is a preadaptation for the ability to grasp prey. Several species were observed using grasping motions in place of the tongue to capture prey.

Sensory modulation and behavioral choice during feeding in the Australian frog, Cyclorana novaehollandiae.

This study investigates how visual and tactile sensory information, as well as biomechanical effects due to differences in physical characteristics of the prey, influence feeding behavior in the frog Cyclorana novaehollandiae. Video motion analysis was used to quantify movement patterns produced when feeding on five prey types (termites, waxworms, crickets, mice and earthworms). Twelve kinematic variables differed significantly among prey types, and twelve variables were correlated with prey characteristics (including mass, length, height and velocity of movement). Results indicate that C. novaehollandiae uses a different strategy to capture each prey type. Visual assessment of prey characteristics appeared to be more important in modulating feeding behavior that tactile cues or biomechanical effects. We propose a hierarchical hypothesis of behavioral choice, in which decisions are based primarily on visual analysis of prey characteristics. In this model, the frogs first choose between jaw prehension and tongue prehension based on prey size. If they have chosen jaw prehension, they next choose between upward or downward head rotation based on length and height of the prey. If they have chosen tongue prehension, they next choose between behavior for fast and slow prey. Final decisions may be the result of behavioral fine tuning based on tactile feedback.

The roles of visual and proprioceptive information during motor program choice in frogs.

Previous studies have shown that leopard frogs, Rana pipiens, use tongue prehension to capture small prey and jaw prehension to capture large prey. After hypoglossal nerve transection, the frogs fail to open their mouths when attempting to feed on small prey, but open their mouths and capture large prey. Here, we investigate how visual information about the prey and proprioceptive information from the tongue interact to influence the motor program choice. Using pieces of earthworm of various sizes, we found that Rana exhibits two different behavior patterns based on prey size. The frogs captured the 1.5-cm prey using tongue prehension, whereas 2.0-cm and larger prey were captured using jaw prehension. After hypoglossal transection, the frogs never opened their mouths when they tried to feed on 1.5-cm prey. When feeding on 3.0-cm and larger prey after transection, they always opened their mouths and captured the prey using jaw prehension. When offered 2.0-cm prey, they alternated randomly between opening and not opening the mouth. Therefore, deafferentation changed the pattern of motor program choice at the behavioral border. This implies that afferents from the tongue interact with visual input to influence motor program choice.

Mechanisms of tongue protraction and narial closure in the marine toad Bufo marinus.

Electromyography, kinematic analysis, muscle stimulation and denervation techniques were used to investigate the muscular mechanisms of narial closure during breathing and of tongue protraction during prey capture in the marine toad Bufo marinus. Toads were video-taped during breathing and feeding under a variety of conditions: before surgery, after unilateral or bilateral denervation of the M. submentalis, and after unilateral or bilateral denervation of the Mm. genioglossus basalis and medialis. Deeply anesthetized toads were video-taped during stimulation of several cranial muscles, and electromyograms were recorded from the M. submentalis during feeding before and after its denervation. Bufo marinus differs from many other anurans in having a relatively long tongue that experiences large accelerations (> 31 g) during protraction. Tongue protraction occurs in two phases: an early phase during which the lingual tip moves upward and forward relative to the mandibular tip as the tongue shortens, and a later phase during which the lingual tip moves downward and forward relative to the mandibular tip as the tongue elongates under its own momentum. Relative to an external reference, the lingual tip follows a straight trajectory from mouth to prey, which depends critically upon precise coordination of tongue and jaw movements. The M. submentalis is necessary for normal tongue protraction during feeding. In contrast, the Mm. genioglossus basalis and medialis are necessary for forward movement of the tongue pad over the symphysis. In B. marinus, a simple anatomical change (elongation of the tongue) has functional consequences (inertial elongation) that profoundly affect the mechanisms of neuromuscular control. Though seldom studied, it seems likely that morphological evolution has had a profound influence on mechanisms of motor control in animals generally.

Mechanism of tongue protraction during prey capture in the spadefoot toad Spea multiplicata (Anura: Pelobatidae).

Recent studies have used muscle denervation experiments to examine the function of muscles during feeding in frogs. By comparing the results of denervation experiments among taxa, it is possible to identify evolutionary changes in muscle function. The purpose of this study was to examine the function of jaw and tongue muscles during prey capture in Spea multiplicata, a representative of the superorder Mesobatrachia. All members of this group possess a disjunct hyoid apparatus. We predicted that Spea would possess a novel mechanism of tongue protraction on the basis of its hyoid morphology. High-speed video motion analysis and muscle denervation were used to study the feeding behavior and mechanism of tongue protraction in Spea. Although Spea possesses a relatively long tongue, its feeding behavior is similar to that of short-tongued frogs of similar body size. Denervation of the m. submentalis had no effect on feeding behavior. When the m. geniohyoideus was denervated, the tongue pad was raised and moved forward slightly, but did not leave the mouth. When the m. genioglossus was denervated, the tongue pad was raised slightly, but no forward movement of the tongue occurred. A similar result was obtained after the mm. genioglossus and geniohyoideus were denervated simultaneously. Thus, both the mm. genioglossus and geniohyoideus are necessary for normal tongue protraction in Spea. In contrast, only the m. genioglossus is necessary for normal tongue protraction in archaeobatrachians and neobatrachians. We hypothesize that the disjunct hyoid is responsible for the greater role of hyoid movement during feeding in mesobatrachians.

Feeding kinematics of phyllomedusine tree frogs.

Previous studies have demonstrated that the phyllomedusine hylids possess highly protrusible tongues, a derived characteristic within the family Hylidae. In the present study, the kinematics of the feeding behavior of a phyllomedusine species, Pachymedusa dacnicolor, was analyzed using high-speed video (180 frames s-1). Its behavior was compared with that of Hyla cinerea, a species with a weakly protrusible tongue. P. dacnicolor exhibits a faster rate of tongue protraction, a longer gape cycle and more variable feeding kinematics than H. cinerea. In addition, the tongue is used in a unique 'fly-swatter' fashion, to pin the prey to the substratum as the frog completes the lunge. The rapid tongue protraction, extended gape cycle and fly-swatter action may have evolved in response to a diet of large, rapidly moving insects. In addition, several duration variables of the feeding cycle were greater for misses than for captures and drops, which suggests that sensory feedback rather than biomechanics controls gape cycle duration.