Musculoskeletal and tendinous details of selected anomalies in the locomotor system of anurans.

Previous studies on anuran anomalies predominantly examine isolated cases or focus on external and skeletal features. Our study analyzes a comprehensive sample collected from 1991 to 2017, examining the muscle-tendon system in 24 anuran species across adult, juvenile, and metamorphic stages. This extensive sample size allows us to investigate consistent anomaly patterns across different developmental stages and anuran families, exploring potential common developmental or genetic factors. Our detailed anatomical examination, encompassing musculature, tendons, and skeletal structures, revealed that 21% of the specimens displayed anomalies, a noteworthy finding considering the extensive sample size and duration of the studied sample. Of these anomalies, 17% affected the locomotor system, predominantly in the upper limbs. Key anomalies included, forelimbs and hindlimbs brachydactyly, rotation in forelimbs, partial kyphotic lordosis, and scoliosis. Notably, the digit 4 in the forelimbs and digits 4 and 5 in the hindlimbs were particularly susceptible to teratogenic effects, indicating possible prolonged exposure during development. Our study also uncovered combinations of anomalies and identified a phenotype similar to Poland syndrome. The findings validate the "Logic of Monsters" (LoMo theory) by Alberch, although the name itself may not be deemed appropriate, showing that developmental disruptions in tetrapods are not random but follow distinct sequences and patterns. The name, while unfortunate, accurately reflects the unusual nature of these developmental anomalies. This contributes to the evolving "Evo-Devo-Path" framework, highlighting the study's importance in understanding developmental disruptions in tetrapods.

Prey capture kinematics of horned frogs (Anura: Ceratophryidae).

Horned frogs, members of the Ceratophryidae family, encompass a group of anurans varying in size and behavior, yet unified by morphological and behavioral traits enabling them to adopt a megalophagous diet (i.e., large prey feeding). Although the group has been the focus of numerous studies, our understanding of its feeding behavior remains limited. In this study, we characterize the feeding mechanism in five species representing the three extant genera of ceratophryid anurans, both in terrestrial and aquatic environments. We also explore the ability of Chacophrys pierottii to adjust feeding behavior based on prey type. Our findings show that all species are capable of wide mouth opening, displaying an asymmetric feeding cycle. While tongue usage is the primary method for capturing prey on land, ceratophryids may use their forelimbs to manipulate prey into their mouths, exhibiting different behavioral patterns. C. pierottii shows modulation of its feeding kinematics and is also capable of some modulation of its feeding in response to prey properties.

Modelling evolutionary tempo and mode using formal morphological spaces and Markov chain principles.

Evolutionary tempo and mode summarize ancient and controversial subjects of theoretical biology such as gradualism, convergence, contingence, trends, and entrenchment. We employed an integrative methodological approach to explore the evolutionary tempo and mode of Lepidosaurian phalangeal formulae (PFs). This approach involves quantifying the frequencies of morphological changes along an evolutionary trajectory. The five meristic characters encoded by PFs are particularly valuable in revealing evolutionary patterns, owing to their discrete nature and extensive documentation in the literature. Based on a pre-existing dataset of PFs from 649 taxa (35 Lepidosauria families, including fossils), from which there exists a unique repertoire of 53 formulations, our approach simultaneously considers phenetic and phylogenetic data. This culminates in a diagram accounting for the phylogenetic dynamic of evolution traversing across different regions of morphospace. The method involves enumerating phenotypical options, reconstructing phenotypes across the phylogeny, projecting phenotypes onto a morphospace, and constructing a flow network from the frequency of evolutionary transitions between unique phenotypic conditions. This approach links Markovian chains and evolutionary trajectories to formally define parameters that describe the underlying transitions of morphological change. Among other results, we found that (a) PF evolution exhibits a clear trend towards reduction in the phalangeal count and that (b) evolutionary change tends to occur significantly between morphologically similar PFs. Notwithstanding, although minor but not trivial, transitions between distant formulas -jumps- occur. Our results support a pluralistic view including stasis, gradualism, and saltationism discriminating their prevalence in a target character evolution.

Antagonistic regulation of homeologous uncx.L and uncx.S genes orchestrates myotome and sclerotome differentiation in the evolutionarily divergent vertebral column of Xenopus laevis.

In anurans, the vertebral column diverges widely from that of other tetrapods; yet the molecular mechanisms underlying its morphogenesis remain largely unexplored. In this study, we investigate the role of the homeologous uncx.L and uncx.S genes in the vertebral column morphogenesis of the allotetraploid frog Xenopus laevis. We initiated our study by cloning the uncx orthologous genes in the anuran Xenopus and determining their spatial expression patterns using in situ hybridization. Additionally, we employed gain-of-function and loss-of-function approaches through dexamethasone-inducible uncx constructs and antisense morpholino oligonucleotides, respectively. Comparative analysis of the messenger RNA sequences of homeologous uncx genes revealed that the uncx.L variant lacks the eh1-like repressor domain. Our spatial expression analysis indicated that in the presomitic mesoderm and somites, the transcripts of uncx.L and uncx.S are located in overlapping domains. Alterations in the function of uncx genes significantly impact the development and differentiation of the sclerotome and myotome, resulting in axial skeleton malformations. Our findings suggest a scenario where the homeologous genes uncx.L and uncx.S exhibit antagonistic functions during somitogenesis. Specifically, uncx.S appears to be crucial for sclerotome development and differentiation, while uncx.L primarily influences myotome development. Postallotetraploidization, the uncx.L gene in X. laevis evolved to lose its eh1-like repressor domain, transforming into a "native dominant negative" variant that potentially competes with uncx.S for the same target genes. Finally, the histological analysis revealed that uncx.S expression is necessary for the correct formation of pedicles and neural arch of the vertebrae, and uncx.L is required for trunk muscle development.

An overview of the osseous palmar sesamoid in Anura, with the particular case of some Rhinella species.

Background: Sesamoids are generally regarded as structures that are not part of the tetrapod body plan. The presence of a palmar sesamoid is assumed to serve as a distribution point for the forces of the flexor digitorum communis muscle to the flexor tendons of the digits, which are embedded in the flexor plate. It has been considered that the palmar sesamoid is present in most anuran groups, and it has been suggested that it acts by inhibiting the closing of the palm, preventing grasping. Typical arboreal anuran groups lack a palmar sesamoid and flexor plate, a pattern shared with other tetrapod groups, which can retain a reduced sesamoid and flexor plate. We focus on the anatomical structure of the Rhinella group, which includes species that present an osseous palmar sesamoid and climb bushes or trees to avoid depredation or escape dangerous situations, and can exhibit scansorial and arboreal behaviors. We also add data on the bony sesamoids of 170 anuran species to study the anatomy and evolution of the osseous palmar sesamoid within this amphibian group. Our objective is to bring an overview of the osseous palmar sesamoid in anurans, unveiling the relationship between this element of the manus, its phylogeny, and the anuran habitat use. Methods: Skeletal whole-mount specimens of Rhinella were cleared and double-dyed to describe the sesamoid anatomy and related tissues. We review and describe the palmar sesamoid of 170 anuran species from CT images downloaded from Morphosource.org, representing almost all Anuran families. We performed an standard ancestral state reconstruction by optimizing two selected characters (osseous palmar sesamoid presence, distal carpal palmar surface) along with the habitat use of the sampled taxa, using parsimony with Mesquite 3.7. Results: Our primary finding is that sesamoid optimization in the anuran phylogeny revealed that its presence is associated with certain clades and not as widespread as previously anticipated. Additionally, we will also be delving into other important outcomes of our study that are relevant to those working in the field of anuran sesamoids. The osseous palmar sesamoid is present in the clade Bufonidae-Dendrobatidae-Leptodactylidae-Brachicephalidae that we named as PS clade, and also in the archeobatrachian pelobatoid Leptobranchium, all strongly terrestrial and burrowing species, though with exceptions. The osseous palmar sesamoid is always present in Bufonidae, but varies in form and size, depending on the mode that they use their manus, such as in the Rhinella margaritifera which has a cylindrical one and also grasping abilities that involve closing the manus. The scattered presence of the bony palmar sesamoid among anuran clades raises the question whether this sesamoid can be present with a different tissular composition in other groups.

Ontogeny of the meniscus in the anuran Xenopus laevis.

The anuran knee joint is subjected to the jump, one of the tetrapods' most demanding mechanical stresses. Consistent with this continuous effort, the knee of the anurans has a complex structure comparable to that of an amniote. Here, we describe the ontogeny of the Xenopus knee tissues and study the morphogenesis of the knee joint shape by performing a geometric morphometric analysis of specially selected anatomical structures: the menisci and the long bone epiphyses. A meniscus is a crescent-shaped fibrocartilaginous structure, with a triangular cross-section inserted between joints surfaces. A meniscus transmits load across the tibiofemoral joint by increasing congruity of the long bone epiphysis and decreasing the resulting stress exerted on the articular cartilage. We ask two questions: (1) what is the tissue composition along the ontogeny of the menisci of a swimming frog? (2) How do the menisci acquire the shape that will allow their adjustment? We studied the structures and tissue ontogeny of the knee of several specimens of Xenopus laevis and evaluated the congruity of the knee structures across the species ontogeny. Histological sections showed that the cavitation process responsible for separating the menisci and the epiphyses seems to be pivotal in shaping the conformity of these structures and the long bone epiphyses of the hindlimbs. The geometric morphometric analysis allowed us to interpret three phases of differentiation associated with limb functionality. The characteristic shape of the meniscus appears early in the ontogeny of the knee, simultaneously with the epiphysis contours.

Insights on the evolution of Coronavirinae in general, and SARS-CoV-2 in particular, through innovative biocomputational resources.

The structural proteins of coronaviruses portray critical information to address issues of classification, assembly constraints, and evolutionary pathways involving host shifts. We compiled 173 complete protein sequences from isolates belonging to the four genera of the subfamily Coronavirinae. We calculate a single matrix of viral distance as a linear combination of protein distances. The minimum spanning tree (MST) connecting the individuals captures the structure of their similarities. The MST re-capitulates the known phylogeny of Coronovirinae. Hosts were mapped onto the MST and we found a non-trivial concordance between host phylogeny and viral proteomic distance. We also study the chimerism in our dataset through computational simulations. We found evidence that structural units coming from loosely related hosts hardly give rise to feasible chimeras in nature. This work offers a fresh way to analyze features of SARS-CoV-2 and related viruses.

Hind limb muscles influence the architectural properties of long bones in frogs.

The Mechanostat Theory states that osteocytes sense both the intensity and directionality of the strains induced by mechanical usage and modulate the bone design accordingly. In long bones, this process may adapt anterior-posterior and lateral-medial strength to their mechanical environment showing regional specificity. Anuran species are ideal for analyzing the muscle-bone relationships related to the different mechanical stresses induced by their many locomotor modes and habitat uses. This work aimed to explore the relationships between indicators of the force of the most relevant muscles to locomotion and the mechanical properties of femur and tibia fibula in preserved samples of three anuran species with different habitat use (aquatic, arboreal) and locomotion modes (swimmer, jumper, walker/climber). For that purpose, we measured the anatomical cross-sectional area of each dissected muscle and correlated it with the moments of inertia and bone strength indices. Significant, species-specific covariations between muscle and bone parameters were observed. Pseudis platensis, the aquatic swimmer, showed the largest muscles, followed by Boana faber, the jumper and Phyllomedusa sauvagii, the walker/climber. As we expected, bigger muscles correlate with bone parameters in all the species. Nevertheless, smaller muscles also play an important role in bone design. In aquatic species, muscle interaction enhances mostly lateral bending strength throughout the femur and lateral and antero-posterior bending strength in the tibia fibula. In the jumper species, muscles affected the femur and tibia fibula mostly in anterior-posterior bending. In the walker/climber species, responses involving both antero-posterior and lateral bending strengths were observed in the femur and tibia fibula. These results show that bones will be more or less resistant to lateral and antero-posterior bending according to the different mechanical challenges of locomotion in aquatic vs. arboreal habitats. This study provides new evidence of the muscle-bone relationships in three frog species associated with their different locomotion and habitat uses, highlighting the crucial role of muscle in determining the architectural properties of bones.

Morphospace analysis leads to an evo-devo model of digit patterning.

Biological forms occupy a constrained portion of theoretical morphospaces. Developmental models accounting for empirical morphospaces are necessary to achieve a better understanding of this phenomenon. We analyzed the phalangeal formulas (PFs) in lizards and relatives' hands by comparing them with a set of simulated PFs that compose a theoretical morphospace. We detected that: (1) the empirical morphospace is severely limited in size, (2) the PFs comply with two properties of phalangeal count per digit, namely the ordering rule (DI ≤ DII ≤ DIII ≤ DIV ≥ DV), and the contiguity relationship (neighbor digits differ on average in one phalanx), (3) the totality of the PFs can be categorized into four categories of hands aligned along a feasibility gradient. We also reconstructed the evolution of PFs and found a stepwise trajectory from the plesiomorphic PF towards reduced conditions. Finally, we propose a developmental model as the generative mechanism behind the PFs. It is consistent with the bulk of evidence managed and involves an ordered digit primordia initialization timed with periodic signals of joint formation coming from digit tips. Our approach is also useful to address the study of other meristic sequences in nature such as dental, floral, and branchial formulas.

Forelimb musculoskeletal-tendinous growth in frogs.

The tendons unite and transmit the strength of the muscles to the bones, allowing movement dexterity, the distribution of the strength of the limbs to the digits, and an improved muscle performance for a wide range of locomotor activities. Tissue differentiation and maturation of the structures involved in locomotion are completed during the juvenile stage; however, few studies have investigated the ontogenetic variation of the musculoskeletal-tendinous system. We ask whether all those integrated tissues and limb structures growth synchronically between them and along with body length. We examined the ontogenetic variation in selected muscles, tendons and bones of the forelimbs in seventy-seven specimens belonging to seven anuran species of different clades and of three age categories, and investigate the relative growth of the forelimb musculoskeletal-tendinous structures throughout ontogeny. Ten muscles and nine tendons and their respective large bones (humerus and radioulna) were removed intact, and their length was measured and analyzed through a multivariate approach of allometry. We obtained an allometry coefficient, which indicates how the coefficient departures from isometry as well as allometric trends. Our data suggest that along with the post-metamorphic ontogeny, muscles tend to elongate proportionally to bone length, with a positive allometric trend. On the contrary, tendons show a negative allometric growth trend. Only two species show different patterns: Rhinella granulosa and Physalaemus biligonigerus, with an isometric and positive growth of muscles and bones, and most tendons being isometric.

Biomechanical properties of anuran long bones: correlations with locomotor modes and habitat use.

Long bones are subjected to mechanical loads during locomotion that will influence their biomechanical properties through a feedback mechanism (the bone mechanostat). This mechanism adapts the spatial distribution of the mineralized tissue to resist compression, bending and torsion. Among vertebrates, anurans represent an excellent group to study long bone properties because they vary widely in locomotor modes and habitat use, which enforce different skeletal loadings. In this study, we hypothesized that (a) the cortical bone mass, density and design of anuran femur and tibiofibula would reflect the mechanical influences of the different locomotor modes and habitat use, and (b) the relationships between the architectural efficiency of cortical design (cross-sectional moments of inertia) and the intrinsic stiffness of cortical tissue [cortical mineral density; the 'distribution/quality' (d/q) relationship] would describe some inter-specific differences in the efficiency of the bone mechanostat to improve bone design under different mechanical loads. To test this hypothesis, we determined tomographic (peripheral quantitative computed tomography) indicators of bone mass, mineralization, and design along the femur and tibiofibula of four anuran species with different modes of locomotion and use of habitat. We found inter-specific differences in all measures between the distal and proximal ends and mid-diaphysis of the bones. In general, terrestrial-hopper species had the highest values. Arboreal-walker species had the lowest values for all variables except for cortical bone mineral density, which was lowest in aquatic-swimmer species. The d/q relationships showed similar responses of bone modeling as a function of cortical stiffness for aquatic and arboreal species, whereas terrestrial-hoppers had higher values for moments of inertia regardless of the tissue compliance to be deformed. These results provide new evidence regarding the significant role of movement and habitat use in addition to the biomechanical properties of long bones within a morpho-functional and comparative context in anuran species.

Sesamoids in Caudata and Gymnophiona (Lissamphibia): absences and evidence.

An integrative definition of sesamoid bones has been recently proposed, highlighting their relationship with tendons and ligaments, their genetic origin, the influence of epigenetic stimuli on their development, and their variable tissue composition. Sesamoid bones occur mainly associated with a large number of mobile joints in vertebrates, most commonly in the postcranium. Here, we present a survey of the distribution pattern of sesamoids in 256 taxa of Caudata and Gymnophiona and 24 taxa of temnospondyls and lepospondyls, based on dissections, high-resolution X-ray computed tomography from digital databases and literature data. These groups have a pivotal role in the interpretation of the evolution of sesamoids in Lissamphibia and tetrapods in general. Our main goals were: (1) to contribute to the knowledge of the comparative anatomy of sesamoids in Lissamphibia; (2) to assess the evolutionary history of selected sesamoids. We formally studied the evolution of the observed sesamoids by optimizing them in the most accepted phylogeny of the group. We identified only three bony or cartilaginous sesamoids in Caudata: the mandibular sesamoid, which is adjacent to the jaw articulation; one located on the mandibular symphysis; and one located in the posterior end of the maxilla. We did not observe any cartilaginous or osseous sesamoid in Gymnophiona. Mapping analyses of the sesamoid dataset of urodeles onto the phylogeny revealed that the very conspicuous sesamoid in the mandibular symphysis of Necturus beyeri and Amphiuma tridactylum is an independent acquisition of these taxa. On the contrary, the sesamoid located between the maxilla and the lower jaw is a new synapomorphy that supports the node of Hydromantes platycephalus and Karsenia coreana. The absence of a mandibular sesamoid is plesiomorphic to Caudata, whereas it is convergent in seven different families. The absence of postcranial sesamoids in salamanders might reveal a paedomorphic pattern that would be visible in their limb joints.

Anuran forelimb muscle tendinous structures and their relationship with locomotor modes and habitat use.

The interaction between organisms and their environment is central in functional morphology. Differences in habitat usage may imply divergent morphology of locomotor systems; thus, detecting which morphological traits are conservative across lineages and which ones vary under environmental pressure is important in evolutionary studies. We studied internal and external morphology in 28 species of Neotropical anurans. Our aim was to determine if internal morphology (muscle and tendons) shows lower phylogenetic signal than external morphology. In addition, we wanted to know if morphology varies in relation to the habitat use and if there are different functional groups. We found differences in the degree of phylogenetic signal on the groups of traits. Interestingly, postaxial regions of the forelimb are evolutionarily more labile than the preaxial regions. Phylomorphospace plots show that arboreal (jumpers and graspers) and swimmer frogs cluster based on length of fingers and the lack of sesamoid, also reflected by the use of habitat. These functional clusters are also related to phylogeny. Sesamoid and flexor plate dimensions together with digit tendons showed to be important to discriminate functional groups as well as use of habitat classification. Our results allow us to identify a "grasping syndrome" in the hand of these frogs, where palmar sesamoid and flexor plate are absent and a third metacarpal with a bony knob are typical. Thus, a lighter skeleton, long fingers and a prensile hand may be key for arboreality.

Functional morphology and identity of the thenar pad in the subterranean genus Ctenomys (Rodentia, Caviomorpha).

As in many other fossorial tetrapods, the most obvious adaptations to scratch-digging in the subterranean tuco-tuco (Rodentia, Ctenomyidae, Ctenomys) are found in the hands, which among other adaptations, present the mesaxonic condition; i.e. the central digits are more developed, and also their claws, which are curved and elongated. The thumb is atrophied and aligned with the rest of the digits, showing a small and flat claw. This configuration of digits and claws seems to be in accordance with what it is expected for rodents: rudimentary movements when handling food items. However, on the palmar side of the hand, tuco-tucos have several pads, the thenar (located under the thumb) being the most developed. In this study, we investigated the functional morphology of the thenar pad through different approaches: musculoskeletal anatomy, histology and functionality. The analysis of radiographs and clarified and double-stained hand samples of Ctenomys talarum and C. australis showed that the thenar pad is supported by a paddle-shaped bone that articulates with a protrusion in the scapholunate bone. This bone, flat and long, continues in a flat cartilaginous structure, with a shape similar to a claw. Dissections showed that the thenar pad has several associated muscles: the m. palmaris longus, the m. abductor pollicis longus, and a massive muscular complex located between the thumb and the thenar pad. By topology it might be inferred that this complex is formed by the m. abductor pollicis brevis, the m. flexor pollicis brevis and the m. adductor pollicis brevis. Longitudinal histological sections of the thenar pad stained with hematoxylin-eosin showed a thick layer of keratin at the distal end, external face. The observation of live specimens of C. talarum foraging on two food items of different size and filmed at 300 fps showed that the thenar pad acts as an opposable thumb, with digit-like movements. Tuco-tucos are able to perform more precise movements than expected, and to grasp and manipulate the food with one hand. In previous studies, it was suggested that the thenar pad was supported by a 'palmar ossicle', or 'prepollex' (= radial sesamoid bone). Our results suggest that this sesamoid underwent a radical change on its morphology, making the thenar pad a part of the food handling system in Ctenomys, so the thenar pad might be considered a 'false thumb', rather than a palmar pad. It is suggested to advance on the description and functional analysis of the thenar pad, redefining the structure, since the terms used so far to define it would not be accurate.

Sesamoids in tetrapods: the origin of new skeletal morphologies.

Along with supernumerary bones, sesamoids, defined as any organized intratendinous/intraligamentous structure, including those composed of fibrocartilage, adjacent to an articulation or joint, have been frequently considered as enigmatic structures associated with the joints of the skeletal system of vertebrates. This review allows us to propose a dynamic model to account for part of skeletal phenotypic diversity: during evolution, sesamoids can become displaced, attaching to and detaching from the long bone epiphyses and diaphysis. Epiphyses, apophyses and detached sesamoids are able to transform into each other, contributing to the phenotypic variability of the tetrapod skeleton. This dynamic model is a new paradigm to delineate the contribution of sesamoids to skeletal diversity. Herein, we first present a historical approach to the study of sesamoids, discussing the genetic versus epigenetic theories of their genesis and growth. Second, we construct a dynamic model. Third, we present a summary of literature on sesamoids of the main groups of tetrapods, including veterinary and human clinical contributions, which are the best-studied aspects of sesamoids in recent decades. Finally, we discuss the identity of certain structures that have been labelled as sesamoids despite insufficient formal testing of homology. We also propose a new definition to help the identification of sesamoids in general. This review is particularly timely, given the recent increasing interest and research activity into the developmental biology and mechanics of sesamoids. With this updated and integrative discussion, we hope to pave the way to improve the understanding of sesamoid biology and evolution.

Morphology and evolution of the wing bullae in South American Leptophlebiidae (Ephemeroptera).

Insects were the first animals to take to the skies, and have been flying for over 320 million years. The order Ephemeroptera is, or at least is part of, the most early-diverging lineage of extant winged insects. The extant species present a very short adult life span, mainly dedicated to reproduction and dispersal of eggs. Mating and egg-laying behavior depend on flight. Wings are structures to fly and as such face a number of physical and physiological challenges. The convex curvature along the anterior-posterior axis of the wing generates a camber that must be carefully regulated. One of the most interesting ways of wing bending is provided by the bullae, which have been defined as short sections of flexible chitin, where the flexion lines cross veins. Although the bullae have been frequently used as taxonomic characters, there is no study focused on their morphology, although their prevalence on the wings of mayflies strongly suggests a role in flight. In order to identify evolutionary trends of these structures within Ephemeroptera, we constructed a matrix with comparative anatomy data of the bullae from whole mounts of the wings of 300 specimens belonging to 70 species of several mayfly families, as well as scanning microscopy samples of selected specimens. We also surveyed the number of bullae and their distribution in the wings of the different species within the South American Leptophlebiidae clade. We optimized the characters onto the latest published phylogeny for Leptophlebiidae.

Movement and joints: effects of overuse on anuran knee tissues.

Movement plays a main role in the correct development of joint tissues. In tetrapods, changes in normal movements produce alterations of such tissues during the ontogeny and in adult stages. The knee-joint is ideal for observing the influence of movement disorders, due to biomechanical properties of its components, which are involved in load transmission. We analyze the reaction of knee tissues under extreme exercise in juveniles and adults of five species of anurans with different locomotor modes. We use anurans as the case study because they undergo great mechanical stress during locomotion. We predicted that (a) knee tissues subjected to overuse will suffer a structural disorganization process; (b) adults will experience deeper morphological changes than juveniles; and (c) morphological changes will be higher in jumpers compared to walkers. To address these questions, we stimulated specimens on a treadmill belt during 2 months. We performed histological analyses of the knee of both treated and control specimens. As we expected, overuse caused structural changes in knee tissues. These alterations were gradual and higher in adults, and similar between jumpers and walkers species. This study represents a first approach to the understanding of the dynamics of anuran knee tissues during the ontogeny, and in relation to locomotion. Interestingly, the alterations found were similar to those observed in anurans subjected to reduced mobility and also to those described in joint diseases (i.e., osteoarthritis and tendinosis) in mammals, suggesting that among tetrapods, changes in movement generate similar responses in the tissues involved.

Comparative anatomy and development of pectoral and pelvic girdles in hylid anurans.

The development of the tetrapod pectoral and pelvic girdles is intimately linked to the proximal segments of the fore- and hindlimbs. Most studies on girdles are osteological and provide little information about soft elements such as muscles and tendons. Moreover, there are few comparative developmental studies. Comparative data gleaned from cleared-and-stained whole mounts and serial histological sections of 10 species of hylid frogs are presented here. Adult skeletal morphology, along with bones, muscles, and connective tissue of both girdles and their association with the proximal portions of the anuran fore- and hindlimbs are described. The data suggest that any similarity could be attributable to the constraints of their ball-and-socket joints, including incorporation of the girdle and stylopodium into a single developmental module. An ancestral state reconstruction of key structures and developmental episodes reveals that several development events occur at similar stages in different species, thereby preventing heterochronic changes. The medial contact of the halves of the pectoral girdle coincides with the emergence of the forelimbs from the branchial chamber and with the total differentiation of the linkage between the axial skeleton and the girdles. The data suggest that morphogenic activity in the anterior dorsal body region is greater than in the posterior one, reflecting the evolutionary sequence of the development of the two girdles in ancient tetrapods. The data also document the profound differences in the anatomy and development of the pectoral and pelvic girdles, supporting the proposal that the pectoral and pelvic girdles are not serially homologous, as was long presumed.

Frog tendon structure and its relationship with locomotor modes.

Tendon collagen fibrils are the basic force-transmitting units of the tendon. Yet, surprisingly little is known about the diversity in tendon anatomy and ultrastructure, and the possible relationships between this diversity and locomotor modes utilized. Our main objectives were to investigate: (a) the ultra-structural anatomy of the tendons in the digits of frogs; (b) the diversity of collagen fibril diameters across frogs with different locomotor modes; (c) the relationship between morphology, as expressed by the morphology of collagen fibrils and tendons, and locomotor modes. To assess the relationship between morphology and the locomotor modes of the sampled taxa we performed a principal component analysis considering body length, fibrillar cross sectional area (CSA) and tendon CSA. A MANOVA showed that differences between species with different locomotor modes were significant with collagen fibril diameter being the discriminating factor. Overall, our data related the greatest collagen fibril diameter to the most demanding locomotor modes, conversely, the smallest collagen fibril CSA and the highest tendon CSA were observed in animals showing a hopping locomotion requiring likely little absorption of landing forces given the short jump distances.