Alligators employ intermetatarsal reconfiguration to modulate plantigrade ground contact

The functional morphology and kinematics of the elbow joint remain relatively understudied in squamates. Previous investigations of lizard elbow morphology and kinematics suggest long-axis rotation (LAR) of the radius and ulna during stance allows the manus to remain pronated during forelimb retraction. Using XROMM (X-ray Reconstruction of Moving Morphology), we explored the range of 3D movements and kinematics of the humerus, radius, and ulna in three adult male Central bearded dragons (Pogona vitticeps) during trackway walking. Our data indicate that the elbow joint of P. vitticeps experiences significant rotations in all three dimensions and that the radius and ulna adduct and rotate laterally on their long axes relative to the elbow joint and to one another during stance. These movements allow the distal ends of the radius and ulna to remain in a configuration necessary for manus pronation. These data support previous inferences that the radius and ulna of lizards move independently at the wrist joint. We suggest that independent LAR of the radius and ulna relative to the elbow joint and to one another may be an ancestral mechanism in lizards and perhaps more broadly across non-avian reptiles.

ABSTRACTThe current hypothesis regarding the mechanics of breathing in crocodylians is that the double-headed ribs, with both a capitulum and tuberculum, rotate about a constrained axis passing through the two articulations; moreover, this axis shifts in the caudal thoracic ribs, as the vertebral parapophysis moves from the centrum to the transverse process. Additionally, the ventral ribcage in crocodylians is thought to possess additional degrees of freedom through mobile intermediate ribs. In this study, X-ray reconstruction of moving morphology (XROMM) was used to quantify rib rotation during breathing in American alligators. Whilst costovertebral joint anatomy predicted overall patterns of motion across the ribcage (decreased bucket handle motion and increased calliper motion), there were significant deviations: anatomical axes overestimated pump handle motion and, generally, ribs in vivo rotate about all three body axes more equally than predicted. The intermediate ribs are mobile, with a high degree of rotation measured about the dorsal intracostal joints, especially in the more caudal ribs. Motion of the sternal ribs became increasingly complex caudally, owing to a combination of the movements of the vertebral and intermediate segments. As the crocodylian ribcage is sometimes used as a model for the ancestral archosaur, these results have important implications for how rib motion is reconstructed in fossil taxa, and illustrate the difficulties in reconstructing rib movement based on osteology alone.

Measuring range of motion (ROM) is a valuable technique that can link bone morphology to joint function in both extant and extinct taxa. ROM results are commonly presented as tables or graphs of maxima and minima for each rotational degree of freedom. We investigate the interactions among three degrees of freedom using X-ray reconstruction of moving morphology (XROMM) to measure ROM of the main hind limb joints of Helmeted Guineafowl (Numida meleagris). By plotting each rotation on an axis, we generate three-dimensional ROM volumes or envelopes composed of hundreds of extreme joint positions for the hip, knee, and intertarsal joints. We find that the shapes of ROM volumes can be quite complex, and that the contribution of long-axis rotation is often substantial. Plotting invivo poses from individual birds executing different behaviors shows varying use of potential rotational combinations within their ROM envelopes. XROMM can provide unprecedented high-resolution data on the spatial relationship of skeletal elements and thereby illuminate/elucidate the complex ways in which soft and hard tissues interact to produce functional joints. In joints with three rotational degrees of freedom, two-dimensional representations of ROM (flexion/extension and abduction/adduction) are incomplete.

Crocodilians have played a significant role in evolutionary studies of archosaurs. Given that several major shifts in forelimb function occur within Archosauria, forelimb morphologies of living crocodilians are of particular importance in assessing locomotor evolutionary scenarios. A previous X-ray investigation of walking alligators revealed substantial movement of the shoulder girdle, but as the sternal cartilages do not show up in X-ray, the source of the mobility could not be conclusively determined. Scapulocoracoid movement was interpreted to indicate independent sliding of each coracoid at the sternocoracoid joint; however, rotations of the sternum could also produce similar displacement of the scapulocoracoids. Here, we present new data employing marker-based XROMM (X-ray reconstruction of moving morphology), wherein simultaneous biplanar X-ray video and surgically implanted radio-opaque markers permit precise measurement of the vertebral axis, sternum and coracoid in walking alligators. We found that movements of the sternum and sternocoracoid joint both contribute to shoulder girdle mobility and stride length, and that the sternocoracoid contribution was less than previously estimated. On average, the joint contributions to stride length (measured with reference to a point on the distal radius, thus excluding wrist motion) are as follows: thoracic vertebral rotation 6.2±3.7%, sternal rotation 11.1±2.5%, sternocoracoid joint 10.1±5.2%, glenohumeral joint 40.1±7.8% and elbow 31.1±4.2%. To our knowledge, this is the first evidence of sternal movement relative to the vertebral column (presumably via rib joints) contributing to stride length in tetrapods.

Crocodylians occupy a key phylogenetic position for investigations of archosaur locomotor evolution. Compared to the well-studied hindlimb, relatively little is known about the skeletal movements and mechanics of the forelimb. In this study, we employed manual markerless XROMM (X-ray Reconstruction Of Moving Morphology) to measure detailed 3-D kinematics of the shoulder girdle and forelimb bones of American alligators (Alligator mississippiensis) walking on a treadmill. Digital models of the interclavicle, scapulocoracoid, humerus, radius and ulna were created using a 3-D laser scanner. Models were articulated and aligned to simultaneously recorded frames of fluoroscopic and standard light video to reconstruct and measure joint motion. Joint coordinate systems were established for the coracosternal, glenohumeral and elbow joints. Our analysis revealed that the limb joints only account for about half of fore/aft limb excursion; the remaining excursion results from shoulder girdle movements and lateral bending of the vertebral column. Considerable motion of each scapulocoracoid relative to the vertebral column is consistent with coracosternal mobility. The hemisellar design of the glenohumeral joint permits some additional translation, or sliding in the fore-aft plane, but this movement does not have much of an effect on the distal excursion of the bone.

Extant archosaurs exhibit highly divergent articular soft tissue anatomies between avian and crocodilian lineages. However, the general lack of understanding of the dynamic interactions among archosaur joint soft tissues has hampered further inferences about the function and evolution of these joints. Here we use contrast-enhanced computed tomography to generate 3D surface models of the pelvis, femora, and hip joint soft tissues in an extant archosaur, the American alligator. The hip joints were then animated using marker-based X-Ray Reconstruction of Moving Morphology (XROMM) to visualize soft tissue articulation during forward terrestrial locomotion. We found that the anatomical femoral head of the alligator travels beyond the cranial extent of the bony acetabulum and does not act as a central pivot, as has been suggested for some extinct archosaurs. Additionally, the fibrocartilaginous surfaces of the alligator's antitrochanter and femoral neck remain engaged during hip flexion and extension, similar to the articulation between homologous structures in birds. Moreover, the femoral insertion of the ligamentum capitis moves dorsoventrally against the membrane-bound portion of the medial acetabular wall, suggesting that the inner acetabular foramen constrains the excursion of this ligament as it undergoes cyclical stretching during the step cycle. Finally, the articular surface of the femoral cartilage model interpenetrates with those of the acetabular labrum and antitrochanter menisci; we interpret such interpenetration as evidence of compressive deformation of the labrum and of sliding movement of the menisci. Our data illustrate the utility of XROMM for studying in vivo articular soft tissue interactions. These results also allow us to propose functional hypotheses for crocodilian hip joint soft tissues, expanding our knowledge of vertebrate connective tissue biology and the role of joint soft tissues in locomotor behavior.

Seasonal variation in plasma thyroxine concentrations in juvenile alligators ( Alligators mississippiensis) from three Florida Lakes

Seasonal variation in plasma sex steroid concentrations in juvenile American alligators

The use of celestial cues for compass orientation was investigated in juvenile American alligators. Animals entrained to new shoreline directions in outdoor training pens moved toward the land end of their y-axis when celestial cues were available. Under overcast skies no significant departure from a uniform circular distribution was apparent. Alligators were able to use solar, stellar or lunar compass mechanisms in this directed movement. The solar compass was time-compensated as indicated by phase-shift tests. Orientation was shifted in the predicted direction after a 6 hour advance in the L:D cycle. The stellar compass was neither time compensated nor bidirectional, as has previously been shown for amphibians, which suggests that the orientation may be based on stellar patterns. Adaptive advantages for celestial compass mechanisms may include predation avoidance, thermoregulation and increased foraging efficiency.

Archosaurs, like all vertebrates, have different types of joints that allow or restrict cranial kinesis, such as synovial joints and fibrous joints. In general, synovial joints are more kinetic than fibrous joints, because the former possess a fluid-filled cavity and articular cartilage that facilitate movement. Even though there is a considerable lack of data on the microstructure and the structure-function relationships in the joints of extant archosaurs, many functional inferences of cranial kinesis in fossil archosaurs have hinged on the assumption that elongated condylar joints are (i) synovial and/or (ii) kinetic. Cranial joint microstructure was investigated in an ontogenetic series of American alligators, Alligator mississippiensis All the presumably synovial, condylar joints found within the head of the American alligator (the jaw joint, otic joint and laterosphenoid-postorbital (LS-PO) joint) were studied by means of paraffin histology and undecalcified histology paired with micro-computed tomography data to better visualize three-dimensional morphology. Results show that among the three condylar joints of A. mississippiensis, the jaw joint was synovial as expected, but the otherwise immobile otic and LS-PO joints lacked a synovial cavity. Therefore, condylar morphology does not always imply the presence of a synovial articulation nor mobility. These findings reveal an undocumented diversity in the joint structure of alligators and show that crocodylians and birds build novel, kinetic cranial joints differently. This complicates accurate identification of synovial joints and functional inferences of cranial kinesis in fossil archosaurs and tetrapods in general.

In evolutionary biomechanics, musculoskeletal computer models of extant and extinct taxa are often used to estimate joint range of motion (ROM) and muscle moment arms (MMAs), two parameters which form the basis of functional inferences. However, relatively few experimental studies have been performed to validate model outputs. Previously, we built a model of the short-beaked echidna (Tachyglossus aculeatus) forelimb using a traditional modelling workflow, and in this study we evaluate its behaviour and outputs using experimental data. The echidna is an unusual animal representing an edge-case for model validation: it uses a unique form of sprawling locomotion, and possesses a suite of derived anatomical features, in addition to other features reminiscent of extinct early relatives of mammals. Here we use diffusible iodine-based contrast-enhanced computed tomography (diceCT) alongside digital and traditional dissection to evaluate muscle attachments, modelled muscle paths, and the effects of model alterations on the MMA outputs. We use X-ray Reconstruction of Moving Morphology (XROMM) to compare ex vivo joint ROM to model estimates based on osteological limits predicted via single-axis rotation, and to calculate experimental MMAs from implanted muscles using a novel geometric method. We also add additional levels of model detail, in the form of muscle architecture, to evaluate how muscle torque might alter the inferences made from MMAs alone, as is typical in evolutionary studies. Our study identifies several key findings that can be applied to future models. 1) A light-touch approach to model building can generate reasonably accurate muscle paths, and small alterations in attachment site seem to have minimal effects on model output. 2) Simultaneous movement through multiple degrees of freedom, including rotations and translation at joints, are necessary to ensure full joint ROM is captured; however, single-axis ROM can provide a reasonable approximation of mobility depending on the modelling objectives. 3) Our geometric method of calculating MMAs is consistent with model-predicted MMAs calculated via partial velocity, and is a potentially useful tool for others to create and validate musculoskeletal models. 4) Inclusion of muscle architecture data can change some functional inferences, but in many cases reinforced conclusions based on MMA alone.

Rarity of soft tissue preservation, including of articular cartilage, in the fossil record hinders creation of biologically-realistic mechanical models. Previous studies of articular cartilage in extant taxa have documented important aspects of cartilage shapes and thicknesses, but these insights remain generalized and have yet to see systematic implementation in biomechanical modeling. Herein, we present a new method for modeling joints that allows for testing of hypotheses about articular cartilage morphology in extinct taxa. Our case study examines the left elbow joint of the sauropod dinosaurDreadnoughtus schraniusing articular cartilage reconstructions constrained by extant phylogenetic bracketing (EPB). EPB investigations of alligator and chicken articular cartilage revealed the presence of a spherical anterior projection of cartilage on the distal humerus which articulates with the radius during flexion. Importantly, this shape does not directly mirror the underlying bone. Using multibody dynamic models created in Adams™ withouta priorirestrictions on joint degrees of freedom, we simulated the effects of three alternative cartilage reconstructions based on these EPB findings which differ in mediolateral placement of a cartilage sphere and its anteroposterior thickness, encompassing a range of possibilities for the condition inDreadnoughtus. Bone kinematics and contact area (calculated in Geomagic®) were tracked. Additionally, we modeled the elbow of an alligator and turkey using the same methodology and compared the results to XROMM (X-ray Reconstruction of Moving Morphology) analysis of the same limbs. Each model produced distinct results but were generally similar supporting our modeling methodology. Based on these findings, we predict thatDreadnoughtus, and presumably other extinct archosaurs, had a spherical projection of cartilage on the anterior face of the distal end of the humerus for articulation with the radius. Though many valuable insights have been gained by existing modeling methodologies, we chose a different approach that focused on joint contact surfaces. Moreover, applying our methods within a quantitative hypothesis-testing framework can advance the field of paleobiology by testing hypotheses relating shape and kinematics that are not possible with prescribed joint motions.

Tongue function is vital for chewing and swallowing and lingual dysfunction is often associated with dysphagia. Better treatment of dysphagia depends on a better understanding of hyolingual morphology, biomechanics, and neural control in humans and animal models. Recent research has revealed significant variation among animal models in morphology of the hyoid chain and suprahyoid muscles which may be associated with variation in swallowing mechanisms. The recent deployment of XROMM (X-ray Reconstruction of Moving Morphology) to quantify 3D hyolingual kinematics has revealed new details on flexion and roll of the tongue during chewing in animal models, movements similar to those used by humans. XROMM-based studies of swallowing in macaques have falsified traditional hypotheses of mechanisms of tongue base retraction during swallowing, and literature review suggests that other animal models may employ a diversity of mechanisms of tongue base retraction. There is variation among animal models in distribution of hyolingual proprioceptors but how that might be related to lingual mechanics is unknown. In macaque monkeys, tongue kinematics—shape and movement—are strongly encoded in neural activity in orofacial primary motor cortex, giving optimism for development of brain–machine interfaces for assisting recovery of lingual function after stroke. However, more research on hyolingual biomechanics and control is needed for technologies interfacing the nervous system with the hyolingual apparatus to become a reality.

Many parameters used to investigate stress in vertebrates are temporally sensitive. The act of capture and sampling can influence them, hindering their functionality for evaluating the effects of stressors. Consequently, the investigation and subsequent incorporation of less time-sensitive parameters are necessary to better evaluate stressors affecting vertebrates. In this study, we investigated how capture stress and handling associated with sampling influence antioxidant status in American Alligators ( Alligator mississippiensis (Daudin, 1802); hereinafter Alligator), long-lived, top-trophic carnivores found in the southeastern United States, by measuring levels of two antioxidant enzymes in destructive (brain and pancreas) and nondestructive (tail scutes) tissues: superoxide dismutase 1 (SOD1) and glutathione peroxidase 1 (GPX1). Capture stress had no effect on pancreatic SOD1 and no effect on brain and pancreatic GPX1 (all p > 0.05). However, brain SOD1, scute SOD1, and scute GPX1 were all impacted by capture stress. These disparate results illustrate that the influence of capture stress on antioxidant enzymes in Alligators is tissue and marker dependent, necessitating further investigation. Our results provide a firm foundation to further investigate oxidative status in crocodilians.

Gene expression patterns in juvenile American alligators ( Alligator mississippiensis) exposed to environmental contaminants