Rolling of the jaw is essential for mammalian chewing and tribosphenic molar function.

Over the past two centuries, mammalian chewing and related anatomical features have been among the most discussed of all vertebrate evolutionary innovations1-3. Chief among these features are two characters: the dentary-only mandible, and the tribosphenic molar with its triangulated upper cusps and lower talonid basin3-5. The flexible mandibular joint and the unfused symphysis of ancestral mammals-in combination with transformations of the adductor musculature and palate-are thought to have permitted greater mobility of each lower jaw, or hemimandible6,7. Following the appearance of precise dental occlusion near the origin of the mammalian crown8,9, therians evolved a tribosphenic molar with a craggy topography that is presumed to have been used to catch, cut and crush food. Here we describe the ancestral tribosphenic therian chewing stroke, as conserved in the short-tailed opossum Monodelphis domestica: it is a simple symmetrical sequence of lower tooth-row eversion and inversion during jaw opening and closing, respectively, enacted by hemimandibular long-axis rotation. This sequence is coupled with an eversion-inversion rotational grinding stroke. We infer that the ancestral therian chewing stroke relied heavily on long-axis rotation, including symmetrical eversion and inversion (inherited from the first mammaliaforms) as well as a mortar-and-pestle rotational grinding stroke that was inherited from stem therians along with the tribosphenic molar. The yaw-dominated masticatory cycle of primates, ungulates and other bunodont therians is derived; it is necessitated by a secondarily fused jaw symphysis, and permitted by the reduction of high, interlocking cusps10-12. The development of an efficient masticatory system-culminating in the tribosphenic apparatus-allowed early mammals to begin the process of digestion by shearing and crushing food into small boli instead of swallowing larger pieces in the reptilian manner, which necessitates a long, slow and wholly chemical breakdown. The vast diversity of mammalian teeth has emerged from the basic tribosphenic groundplan13.

Epaxial and hypaxial co-contraction: a mechanism for modulating strike pressure and accuracy during suction feeding in channel catfish.

Most fish species use concentric epaxial and hypaxial contractions to suction feed, whereby both muscle groups produce cranial expansion and negative intraoral pressures. In contrast, channel catfish (Ictalurus punctatus) suction feed with little to no cranial elevation and epaxial shortening, generating suction power primarily with hypaxial shortening and pectoral girdle retraction. We hypothesized that channel catfish (1) actively anchor the head via isometric contraction of the epaxials and (2) vary feeding performance by modulating the absolute and relative outputs of the co-contracting muscles. We used a combination of electromyography, intraoral pressure recordings and specimen manipulation, and developed a new dual-lever model to explore this idea. We detected epaxial and hypaxial co-contraction prior to suction force development in all strikes. Our model revealed that the differential between the co-contracting muscles may be used to modulate suction pressure and strike accuracy.

Relaxed Feeding Constraints Facilitate the Evolution of Mouthbrooding in Neotropical Cichlids.

AbstractMultifunctionality is often framed as a core constraint of evolution, yet many evolutionary transitions involve traits taking on additional functions. Mouthbrooding, a form of parental care where offspring develop inside a parent's mouth, increases multifunctionality by adding a major function (reproduction) to a structure already serving other vital functions (feeding and respiration). Despite increasing multifunctionality, mouthbrooding has evolved repeatedly from other forms of parental care in at least seven fish families. We hypothesized that mouthbrooding is more likely to evolve in lineages with feeding adaptations that are already advantageous for mouthbrooding. We tested this hypothesis in Neotropical cichlids, where mouthbrooding has evolved four or five times, largely within winnowing clades, providing several pairwise comparisons between substrate-brooding and mouthbrooding sister taxa. We found that the mouthbrooding transition rate was 15 times higher in winnowing than in nonwinnowing clades and that mouthbrooders and winnowers overlapped substantially in their buccal cavity morphologies, which is where offspring are incubated. Species that exhibit one or both of these behaviors had larger, more curved buccal cavities, while species that exhibit neither behavior had narrow, cylindrical buccal cavities. Given the results we present here, we propose a new conceptual model for the evolution of mouthbrooding, integrating the roles of multifunctional morphology and the environment. We suggest that functional transitions like mouthbrooding offer a different perspective on multifunctionality: increasing constraints in one trait may release them for another, generating new evolutionary opportunities.

A new conceptual framework for the musculoskeletal biomechanics and physiology of ray-finned fishes.

Suction feeding in ray-finned fishes requires substantial muscle power for fast and forceful prey capture. The axial musculature located immediately behind the head has been long known to contribute some power for suction feeding, but recent XROMM and fluoromicrometry studies found nearly all the axial musculature (over 80%) provides effectively all (90-99%) of the power for high-performance suction feeding. The dominance of axial power suggests a new framework for studying the musculoskeletal biomechanics of fishes: the form and function of axial muscles and bones should be analysed for power production in feeding (or at least as a compromise between swimming and feeding), and cranial muscles and bones should be analysed for their role in transmitting axial power and coordinating buccal expansion. This new framework is already yielding novel insights, as demonstrated in four species for which suction power has now been measured. Interspecific comparisons suggest high suction power can be achieved in different ways: increasing the magnitude of suction pressure or the rate of buccal volume change, or both (as observed in the most powerful of these species). Our framework suggests that mechanical and evolutionary interactions between the head and the body, and between the swimming and feeding roles of axial structures, may be fruitful areas for continued study.

Royal knifefish generate powerful suction feeding through large neurocranial elevation and high epaxial muscle power.

Suction feeding in ray-finned fishes involves powerful buccal cavity expansion to accelerate water and food into the mouth. Previous XROMM studies in largemouth bass (Micropterus salmoides), bluegill sunfish (Lepomis macrochirus) and channel catfish (Ictalurus punctatus) have shown that more than 90% of suction power in high performance strikes comes from the axial musculature. Thus, the shape of the axial muscles and skeleton may affect suction feeding mechanics. Royal knifefish (Chitala blanci) have an unusual postcranial morphology, with a ventrally flexed vertebral column and relatively large mass of epaxial muscle. Based on their body shape, we hypothesized that royal knifefish would generate high power strikes by utilizing large neurocranial elevation, vertebral column extension and epaxial shortening. As predicted, C. blanci generated high suction expansion power compared with the other three species studied to date (up to 160 W), which was achieved by increasing both the rate of volume change and the intraoral subambient pressure. The large epaxial muscle (25% of body mass) shortened at high velocities to produce large neurocranial elevation and vertebral extension (up to 41 deg, combined), as well as high muscle mass-specific power (up to 800 W kg-1). For the highest power strikes, axial muscles generated 95% of the power, and 64% of the axial muscle mass consisted of the epaxial muscles. The epaxial-dominated suction expansion of royal knifefish supports our hypothesis that postcranial morphology may be a strong predictor of suction feeding biomechanics.

Modular lung ventilation in Boa constrictor.

The evolution of constriction and of large prey ingestion within snakes are key innovations that may explain the remarkable diversity, distribution and ecological scope of this clade, relative to other elongate vertebrates. However, these behaviors may have simultaneously hindered lung ventilation such that early snakes may have had to circumvent these mechanical constraints before those behaviors could evolve. Here, we demonstrate that Boa constrictor can modulate which specific segments of ribs are used to ventilate the lung in response to physically hindered body wall motions. We show that the modular actuation of specific segments of ribs likely results from active recruitment or quiescence of derived accessory musculature. We hypothesize that constriction and large prey ingestion were unlikely to have evolved without modular lung ventilation because of their interference with lung ventilation, high metabolic demands and reliance on sustained lung convection. This study provides a new perspective on snake evolution and suggests that modular lung ventilation evolved during or prior to constriction and large prey ingestion, facilitating snakes' remarkable radiation relative to other elongate vertebrates.

Motor control in the epaxial musculature of bluegill sunfish in feeding and locomotion.

Fishes possess an impressive repertoire of feeding and locomotor behaviors that in many cases rely on the same power source: the axial musculature. As both functions employ different skeletal systems, head versus body, integrating these functions would likely require modular motor control. Although there have been many studies of motor control in feeding or locomotion in fishes, only one study to date has examined both functions in the same individuals. To characterize bilateral motor control of the epaxial musculature in feeding and locomotion, we measured muscle activity and shortening in bluegill sunfish (Lepomis macrochirus) using electromyography and sonomicrometry. We found that sunfish recruit epaxial regions in a dorsal-to-ventral manner to increase feeding performance, such that high-performance feeding activates all the epaxial musculature. In comparison, sunfish seemed to activate all three epaxial regions irrespective of locomotor performance. Muscle activity was present on both sides of the body in nearly all feeding and locomotor behaviors. Feeding behaviors used similar activation intensities on the two sides of the body, whereas locomotor behaviors consistently used higher intensities on the side undergoing muscle shortening. In all epaxial regions, fast-starts used the highest activation intensities, although high-performance suction feeding occasionally showed near-maximal intensity. Finally, active muscle volume was positively correlated with the peak rate of body flexion in feeding and locomotion, indicating a continuous relationship between recruitment and performance. A comparison of these results with recent work on largemouth bass (Micropterus salmoides) suggests that centrarchid fishes use similar motor control strategies for feeding, but interspecific differences in peak suction-feeding performance are determined by active muscle volume.

The Multi-Scale, Three-Dimensional Nature of Skeletal Muscle Contraction.

Muscle contraction is a three-dimensional process, as anyone who has observed a bulging muscle knows. Recent studies suggest that the three-dimensional nature of muscle contraction influences its mechanical output. Shape changes and radial forces appear to be important across scales of organization. Muscle architectural gearing is an emerging example of this process.

Dual function of epaxial musculature for swimming and suction feeding in largemouth bass.

The axial musculature of many fishes generates the power for both swimming and suction feeding. In the case of the epaxial musculature, unilateral activation bends the body laterally for swimming, and bilateral activation bends the body dorsally to elevate the neurocranium for suction feeding. But how does a single muscle group effectively power these two distinct behaviours? Prior electromyographic (EMG) studies have identified fishes' ability to activate dorsal and ventral epaxial regions independently, but no studies have directly compared the intensity and spatial activation patterns between swimming and feeding. We measured EMG activity throughout the epaxial musculature during swimming (turning, sprinting, and fast-starts) and suction feeding (goldfish and pellet strikes) in largemouth bass (Micropterus salmoides). We found that swimming involved obligate activation of ventral epaxial regions whereas suction feeding involved obligate activation of dorsal epaxial regions, suggesting regional specialization of the epaxial musculature. However, during fast-starts and suction feeding on live prey, bass routinely activated the whole epaxial musculature, demonstrating the dual function of this musculature in the highest performance behaviours. Activation intensities in suction feeding were substantially lower than fast-starts which, in conjunction with suboptimal shortening velocities, suggests that bass maximize axial muscle performance during locomotion and underuse it for suction feeding.

Fishes can use axial muscles as anchors or motors for powerful suction feeding.

Some fishes rely on large regions of the dorsal (epaxial) and ventral (hypaxial) body muscles to power suction feeding. Epaxial and hypaxial muscles are known to act as motors, powering rapid mouth expansion by shortening to elevate the neurocranium and retract the pectoral girdle, respectively. However, some species, like catfishes, use little cranial elevation. Are these fishes instead using the epaxial muscles to forcefully anchor the head, and if so, are they limited to lower-power strikes? We used X-ray imaging to measure epaxial and hypaxial length dynamics (fluoromicrometry) and associated skeletal motions (XROMM) during 24 suction feeding strikes from three channel catfish (Ictalurus punctatus). We also estimated the power required for suction feeding from oral pressure and dynamic endocast volume measurements. Cranial elevation relative to the body was small (<5 deg) and the epaxial muscles did not shorten during peak expansion power. In contrast, the hypaxial muscles consistently shortened by 4-8% to rotate the pectoral girdle 6-11 deg relative to the body. Despite only the hypaxial muscles generating power, catfish strikes were similar in power to those of other species, such as largemouth bass (Micropterus salmoides), that use epaxial and hypaxial muscles to power mouth expansion. These results show that the epaxial muscles are not used as motors in catfish, but suggest they position and stabilize the cranium while the hypaxial muscles power mouth expansion ventrally. Thus, axial muscles can serve fundamentally different mechanical roles in generating and controlling cranial motion during suction feeding in fishes.

Channel catfish use higher coordination to capture prey than to swallow.

When animals move they must coordinate motion among multiple parts of the musculoskeletal system. Different behaviours exhibit different patterns of coordination, however, it remains unclear what general principles determine the coordination pattern for a particular behaviour. One hypothesis is that speed determines coordination patterns as a result of differences in voluntary versus involuntary control. An alternative hypothesis is that the nature of the behavioural task determines patterns of coordination. Suction-feeding fishes have highly kinetic skulls and must coordinate the motions of over a dozen skeletal elements to draw fluid and prey into the mouth. We used a dataset of intracranial motions at five cranial joints in channel catfish ( Ictalurus punctatus), collected using X-ray reconstruction of moving morphology, to test whether speed or task best explained patterns of coordination. We found that motions were significantly more coordinated (by 20-29%) during prey capture than during prey transport, supporting the hypothesis that the nature of the task determines coordination patterns. We found no significant difference in coordination between low- and high-speed motions. We speculate that capture is more coordinated to create a single fluid flow into the mouth while transport is less coordinated so that the cranial elements can independently generate multiple flows to reposition prey. Our results demonstrate the benefits of both higher and lower coordination in animal behaviours and the potential of motion analysis to elucidate motor tasks.

Skeletal kinematics of the hyoid arch in the suction-feeding shark Chiloscyllium plagiosum.

White-spotted bamboo sharks, Chiloscyllium plagiosum, generate strong suction-feeding pressures that rival the highest levels measured in ray-finned fishes. However, the hyostylic jaw suspension of these sharks is fundamentally different from the actinopterygian mechanism, including more mobile hyomandibulae, with the jaws and ceratohyal suspended from the hyomandibulae. Prior studies have proposed skeletal kinematics during feeding in orectolobid sharks from indirect measurements. Here, we tested these hypotheses using XROMM to measure cartilage motions directly. In agreement with prior hypotheses, we found extremely large retraction and depression of the ceratohyal, facilitated by large protraction and depression of the hyomandibula. Somewhat unexpectedly, XROMM also showed tremendous long-axis rotation (LAR) of both the ceratohyal and hyomandibula. This LAR likely increases the range of motion for the hyoid arch by keeping the elements properly articulated through their large arcs of motion. XROMM also confirmed that upper jaw protraction occurs before peak gape, similarly to actinopterygian suction feeders, but different from most other sharks in which jaw protrusion serves primarily to close the mouth. Early jaw protraction results from decoupling the rotations of the hyomandibula, with much of protraction occurring before peak gape with the other rotations lagging behind. In addition, the magnitudes of retraction and protraction of the hyoid elements are independent of the magnitude of depression, varying the shape of the mouth among feeding strikes. Hence, the large variation in suction-feeding behavior and performance may contribute to the wide dietary breadth of bamboo sharks.

Shearing overbite and asymmetrical jaw motions facilitate food breakdown in a freshwater stingray, Potamotrygon motoro.

Many species of fish process their prey with cyclic jaw motions that grossly resemble those seen in mammalian mastication, despite starkly different tooth and jaw morphologies. The degree of similarity between the processing behaviors of these disparate taxa has implications for our understanding of convergence in vertebrate feeding systems. Here, we used XROMM (X-ray reconstruction of moving morphology) to investigate prey processing behavior of Potamotrygon motoro, the ocellate river stingray, which has recently been found to employ asymmetrical, shearing jaw motions to break down its prey. We found that P. motoro modulates its feeding kinematics to produce two distinct types of chew cycles: compressive cycles and overbite cycles. The latter are characterized by over-rotation of the upper jaw relative to the lower jaw, past the expected occlusal limit, and higher levels of bilateral asymmetry as compared with compressive chews. We did not find evidence of the mediolateral shearing motions typical of mammalian mastication, but overbite cycles appear to shear the prey item between the upper and lower toothplates in a propalinal fashion. Additionally, comparison of hyomandibular and jaw motions demonstrates that the angular cartilages decouple jaw displacement from hyomandibular displacement in rostrocaudal and mediolateral directions. The multiple similarities between mammalian mastication and the dynamic processing behavior of P. motoro support the use of sub-family Potamotrygoninae as a model for studying evolutionary convergence of mastication-like processing.

XROMM kinematics of ventilation in wild turkeys (Meleagris gallopavo).

The avian ribcage is derived relative to other amniotes, and is hypothesised to be constrained in its movements during ventilation. The double-headed ribs form two articulations with the vertebrae, and are thought to rotate about a strict anatomical axis. However, this costovertebral joint constraint has not been demonstrated empirically and was not found in other taxa with double-headed ribs (i.e. crocodilians). Here, we used X-ray reconstruction of moving morphology (XROMM) to quantify rib rotation in wild turkeys (Meleagris gallopavo) during breathing. We demonstrate that, as predicted from anatomy, the ribs do rotate in a hinge-like manner about a single axis. There is also evidence for elliptical motion of the sternum, as has been reported in other taxa. The evolution of the avian ribcage is closely related to the co-evolution of ventilation and flight, and these results are important for how we model ventilation mechanics in living and fossil birds.

Pectoral and pelvic girdle rotations during walking and swimming in a semi-aquatic turtle: testing functional role and constraint.

Pectoral and pelvic girdle rotations play a substantial role in enhancing stride length across diverse tetrapod lineages. However, the pectoral and pelvic girdle attach the limbs to the body in different ways and may exhibit dissimilar functions, especially during locomotion in disparate environments. Here, we tested for functional differences between the forelimb and hindlimb of the freshwater turtle Pseudemys concinna during walking and swimming using X-ray reconstruction of moving morphology (XROMM). In doing so, we also tested the commonly held notion that the shell constrains girdle motion in turtles. We found that the pectoral girdle exhibited greater rotations than the pelvic girdle on land and in water. Additionally, pelvic girdle rotations were greater on land than in water, whereas pectoral girdle rotations were similar in the two environments. These results indicate that although the magnitude of pelvic girdle rotations depends primarily on whether the weight of the body must be supported against gravity, the magnitude of pectoral girdle rotations likely depends primarily on muscular activity associated with locomotion. Furthermore, the pectoral girdle of turtles rotated more than has been observed in other taxa with sprawling postures, showing an excursion similar to that of mammals (∼38 deg). These results suggest that a rigid axial skeleton and internally positioned pectoral girdle have not constrained turtle girdle function, but rather the lack of lateral undulations in turtles and mammals may contribute to a functional convergence whereby the girdle acts as an additional limb segment to increase stride length.

Intra-oropharyngeal food transport and swallowing in white-spotted bamboo sharks.

Despite the importance of intraoral food transport and swallowing, relatively few studies have examined the biomechanics of these behaviors in non-tetrapods, which lack a muscular tongue. Studies show that elasmobranch and teleost fishes generate water currents as a 'hydrodynamic tongue' that presumably transports food towards and into the esophagus. However, it remains largely unknown how specific musculoskeletal motions during transport correspond to food motion. Previous studies of white-spotted bamboo sharks (Chiloscyllium plagiosum) hypothesized that motions of the hyoid, branchial arches and pectoral girdle, generate caudal motion of the food through the long oropharynx of modern sharks. To test these hypotheses, we measured food and cartilage motion with XROMM during intra-oropharyngeal transport and swallowing (N=3 individuals, 2-3 trials per individual). After entering the mouth, food does not move smoothly toward the esophagus, but rather moves in distinct steps with relatively little retrograde motion. Caudal food motion coincides with hyoid elevation and a closed mouth, supporting earlier studies showing that hyoid motion contributes to intra-oropharyngeal food transport by creating caudally directed water currents. Little correspondence between pectoral girdle and food motion was found, indicating minimal contribution of pectoral girdle motion. Transport speed was fast as food entered the mouth, slower and step-wise through the pharyngeal region and then fast again as it entered the esophagus. The food's static periods in the step-wise motion and its high velocity during swallowing could not be explained by hyoid or girdle motion, suggesting these sharks may also use the branchial arches for intra-oropharyngeal transport and swallowing.

Breathing with floating ribs: XROMM analysis of lung ventilation in savannah monitor lizards.

The structures and functions of the vertebrate lung and trunk are linked through the act of ventilation, but the connections between these structures and functions are poorly understood. We used X-ray reconstruction of moving morphology (XROMM) to measure rib kinematics during lung ventilation in three savannah monitor lizards (Varanus exanthematicus). All of the dorsal ribs, including the floating ribs, contributed to ventilation; the magnitude and kinematic pattern showed no detectable cranial-to-caudal gradient. The true ribs acted as two rigid bodies connected by flexible cartilage, with the vertebral rib and ventromedial shaft of each sternal rib remaining rigid and the cartilage between them forming a flexible intracostal joint. Rib rotations can be decomposed into bucket handle rotation around a dorsoventral axis, pump handle rotation around a mediolateral axis and caliper motion around a craniocaudal axis. Dorsal rib motion was dominated by roughly equal contributions of bucket and pump rotation in two individuals and by bucket rotation in the third individual. The recruitment of floating ribs during ventilation in monitor lizards is strikingly different from the situation in iguanas, where only the first few true ribs contribute to breathing. This difference may be related to the design of the pulmonary system and life history traits in these two species. Motion of the floating ribs may maximize ventilation of the caudally and ventrolaterally positioned compliant saccular chambers in the lungs of varanids, while restriction of ventilation to a few true ribs may maximize crypsis in iguanas.

Bluegill sunfish use high power outputs from axial muscles to generate powerful suction-feeding strikes.

Suction-feeding fish rapidly expand the mouth cavity to generate high-velocity fluid flows that accelerate food into the mouth. Such fast and forceful suction expansion poses a challenge, as muscle power is limited by muscle mass and the muscles in fish heads are relatively small. The largemouth bass powers expansion with its large body muscles, with negligible power produced by the head muscles (including the sternohyoideus). However, bluegill sunfish - with powerful strikes but different morphology and feeding behavior - may use a different balance of cranial and axial musculature to power feeding and different power outputs from these muscles. We estimated the power required for suction expansion in sunfish from measurements of intraoral pressure and rate of volume change, and measured muscle length and velocity. Unlike largemouth bass, the sternohyoideus did shorten to generate power, but it and other head muscles were too small to contribute more than 5-10% of peak expansion power in sunfish. We found no evidence of catapult-style power amplification. Instead, sunfish powered suction feeding by generating high power outputs (up to 438 W kg-1) from their axial muscles. These muscles shortened across the cranial half of the body as in bass, but at faster speeds that may be nearer the optimum for power production. Sunfish were able to generate strikes of the same absolute power as bass, but with 30-40% of the axial muscle mass. Thus, species may use the body and head muscles differently to meet the requirements of suction feeding, depending on their morphology and behavior.

Waddle and shuffle: gait alterations associated with domestication in turkeys.

Domestication has altered turkey morphology by artificially selecting for increased muscle mass and breast meat. Artificial selection has resulted in birds that weigh up to 3 times more than their wild counterparts, with relatively little change in the length of their bones and limbs. Considering these structural changes, it seems probable that domestic turkey locomotor kinematics and kinetics would also be altered. To examine the locomotor dynamics of wild and domestic turkeys, we had both strains walk down a runway with a force plate at the center to measure their ground reaction forces and gait parameters. The location of their center of mass was also quantified using a force plate and bi-planar x-ray and found to be further anterior in the domestic strain. The domestic turkeys locomoted across a lower range of speeds (0.25-1.64 ms-1) than the wild turkeys (0.26-3.26 ms-1) and increased their stride frequency at a higher rate. They also displayed large lateral oscillations, i.e. waddling, during walking that translated into relatively high medio-lateral ground reaction forces and lateral kinetic energy (3.5 times higher than that of wild turkeys). The results indicate that domestic turkey locomotion is not simply a slowed down version of wild turkey locomotion. The changes in gait observed are similar to the shuffling gait present in some human populations, such as Parkinson's patients, which serves to increase stability. The domestic turkey's increased body mass and more anterior center of mass position may require these kinematic and kinetic gait differences.