Skull Anatomy and Ontogeny of Legless Lizard Pseudopus apodus (Pallas, 1775): Heterochronic Influences on Form

Pseudopus apodus (Pallas, 1775) is the largest extant legless species of the subfamily Anguinae (Anguimorpha, Anguidae) living mostly in the sub‐arid territories ranging from the Balkan area in Europe to Kazakhstan in Asia. The species of other two genera live in North America, South‐East Asia and North Africa (Ophisaurus) and Europe and South‐West Asia (Anguis). The interrelationships of Anguinae are unresolved; this is in part the consequence of the insufficient knowledge of the cranial, postcranial and integumentary anatomy of the individual anguine species. The aim of this article is to fulfill this gap in our knowledge of the anguine anatomy. Now, in the first part of the project, the individual bones of the exocranium and visceral endocranium of the anguine legless lizard P. apodus are described in detail. In the present study, P. apodus is revealed to have autoapomorphic features of the skull which clearly distinguish it from Anguis and Ophisaurus. In addition, the study of posthatchling ontogeny of exocranium of P. apodus revealed some features, such as a nasal process of premaxilla being slightly widened in about its mid‐length, that are also typical for adults of the Ophisaurus and Anguis species as well as extinct species of Pseudopus. This strongly indicates that peramorphic heterochronic process played role in the evolution of the P. apodus skull. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 300:460–502, 2017. © 2016 Wiley Periodicals, Inc.     

Limb reduction in squamate reptiles correlates with the reduction of the chondrocranium: A case study on serpentiform anguids

Background

In vertebrates, the skull evolves from a complex network of dermal bones and cartilage—the latter forming the pharyngeal apparatus and the chondrocranium. Squamates are particularly important in this regard as they maintain at least part of the chondrocranium throughout their whole ontogeny until adulthood. Anguid lizards represent a unique group of squamates, which contains limbed and limbless forms and show conspicuous variation of the adult skull.

Results

Based on several emboadryonic stages of the limbless lizards Pseudopus apodus and Anguis fragilis, and by comparing with other squamates, we identified and interpreted major differences in chondrocranial anatomy. Among others, the most important differences are in the orbitotemporal region. P. apodus shows a strikingly similar development of this region to other squamates. Unexpectedly, however, A. fragilis differs considerably in the composition of the orbitotemporal region. In addition, A. fragilis retains a paedomorphic state of the nasal region.

Conclusions

Taxonomic comparisons indicate that even closely related species with reduced limbs show significant differences in chondrocranial anatomy. The Pearson correlation coefficient suggests strong correlation between chondrocranial reduction and limb reduction. We pose the hypothesis that limb reduction could be associated with the reduction in chondrocrania by means of genetic mechanisms.

     

Comparative anatomy and ontogeny of appendicular skeleton of Pseudopus apodus (Pallas, 1775) (Anguimorpha,Anguidae) and a pattern of hindlimb loss in Anguinae

We present a detailed anatomy of the pectoral girdles, pelvic girdles, and hindlimbs of adult and juvenile specimens of Pseudopus apodus (Pallas, 1775). We compared the individual bones of the appendicular skeleton of P. apodus with those of Anguis fragilis and species of Ophisaurus living in North America, North Africa, and Southeast Asia. We found no anatomical features in P. apodus in common with the species of Ophisaurus living in only North America, North Africa, and Southeast Asia. Additionally, we present the prehatching ontogeny of the pelvic girdle of P. apodus and A. fragilis and the prehatching ontogeny of the hindlimb of P. apodus. In the ontogeny of the pelvic girdle of P. apodus, it is possible to distinguish the ossification centers of ilium, ischium, and pubis. In contrast, in the ontogeny of A. fragilis, no ossification centers of ilium, ischium, and pubis are present, and no hindlimb element was detected. In Stage 1 of ontogeny in Pseudopus, the femur and tibia are present; in Stage 2, the nodule representing the fibula appears; and in Stage 3, in addition to the femur, tibia, and fibula, four tarsal elements are present. This anatomical condition corresponds to the anatomical composition of the hindlimb of the adult O. koellikeri. In Stage 4, the involution of all tarsal elements and fibula begins, and in the last two prehatching stages, only femur and tibia remain; this condition is present not only in the adults of Pseudopus, but also in those of several other species of Ophisaurus.     

Reinterpretation of tuberculate cervical vertebrae of Eocene birds as an exceptional anti-predator adaptation against the mammalian craniocervical killing bite

We report avian cervical vertebrae from the Quercy fissure fillings in France, which are densely covered with villi-like tubercles. Two of these vertebrae stem from a late Eocene site, another lacks exact stratigraphic data. Similar cervical vertebrae occur in avian species from Eocene fossils sites in Germany and the United Kingdom, but the new fossils are the only three-dimensionally preserved vertebrae with pronounced surface sculpturing. So far, the evolutionary significance of this highly bizarre morphology, which is unknown from extant birds, remained elusive, and even a pathological origin was considered. We note the occurrence of similar structures on the skull of the extant African rodent Lophiomys and detail that the tubercles represent true osteological features and characterize a distinctive clade of Eocene birds (Perplexicervicidae). Micro-computed tomography (μCT) shows the tubercles to be associated with osteosclerosis of the cervical vertebrae, which have a very thick cortex and much fewer trabecles and pneumatic spaces than the cervicals of most extant birds aside from some specialized divers. This unusual morphology is likely to have served for strengthening the vertebral spine in the neck region, and we hypothesize that it represents an anti-predator adaptation against the craniocervical killing bite (“neck bite”) that evolved in some groups of mammalian predators. Tuberculate vertebrae are only known from the Eocene of Central Europe, which featured a low predation pressure on birds during that geological epoch, as is evidenced by high numbers of flightless avian species. Strengthening of the cranialmost neck vertebrae would have mitigated attacks by smaller predators with weak bite forces, and we interpret these vertebral specializations as the first evidence of “internal bony armor” in birds.     

Comparative osteology of the hynobiid complex Liua-Protohynobius-Pseudohynobius (Amphibia,Urodela): Ⅰ. Cranial anatomy of Pseudohynobius

Hynobiidae are a clade of salamanders that diverged early within the crown radiation and that retain a considerable number of features plesiomorphic for the group. Their evolutionary history is informed by a fossil record that extends to the Middle Jurassic Bathonian time. Our understanding of the evolution within the total group of Hynobiidae has benefited considerably from recent discoveries of stem hynobiids but is constrained by inadequate anatomical knowledge of some extant forms. Pseudohynobius is a derived hynobiid clade consisting of five to seven extant species living endemic to southwestern China. Although this clade has been recognized for over 37 years, osteological details of these extant hynobiids remain elusive, which undoubtedly has contributed to taxonomic controversies over the hynobiid complex Liua-Protohynobius-Pseudohynobius. Here we provide a bone-by-bone study of the cranium in the five extant species of Pseudohynobius (Ps. flavomaculatus, Ps. guizhouensis, Ps. jinfo, Ps. kuankuoshuiensis and Ps. shuichengensis) based on x-ray computer tomography data for 18 specimens. Our results indicate that the cranium in each of these species has a combination of differences in morphology, proportions and articulation patterns in both dermal and endochondral bones. Our study establishes a range of intraspecific differences that will serve as organizing hypotheses for future studies as more extensive collections of these species become available. Morphological features in the cranium for terrestrial ecological adaptation in Hynobiidae are summarized. Based on the results, we also discuss the evolution and development of several potential synapomorphies of Hynobiidae, including features of the orbitosphenoid and articular.     

Effects of Precaudal Elongation on Visceral Topography in a Basal Clade of Ray‐Finned Fishes

Elongate body forms have evolved numerous times independently within Vertebrata. Such body forms have evolved in large part via changes to the vertebral column, either through addition or lengthening of vertebrae. Previous studies have shown that body elongation in fishes has evolved most frequently through the addition of caudal vertebrae. In contrast, however, body elongation in Polypteriformes, a basal clade of ray‐finned fishes (Actinopterygii), has evolved through the addition of precaudal vertebrae; one genus, Erpetoichthys, has approximately twice as many precaudal vertebrae as do members of its sister genus, Polypterus. Thus, polypteriform fishes provide an excellent opportunity to study the effects of precaudal elongation on the gross morphology and organization of visceral organs contained within the body cavity. In this study, we document the anteroposterior positions of most major visceral organs in representative species of both genera (E. calabaricus and P. palmas), relative to both vertebral number and percent pre‐anal length. We found that, whereas the positions of the anterior and posterior borders of the visceral organs relative to percent pre‐anal length were generally similar between the two species, most visceral organs were positioned further posteriorly in E. calabaricus than in P. palmas with respect to vertebral number. Based on previous determinations of the molecular control of anteroposterior patterning of the visceral organs, we discuss which possible changes in gene expression may have led to the anatomical modifications seen in the visceral morphology of Erpetoichthys. Anat Rec, 2012. © 2011 Wiley Periodicals, Inc.     

Differentiation and classification of thoracolumbar transitional vertebrae

The literature states that transitional vertebrae at any junction are characterized by features retained from two adjacent regions in the vertebral column. Currently, there is no published literature available that describes the prevalence or morphology of thoracolumbar transitional vertebrae (TLTV). The aim of this study was to identify the qualitative characteristics of transitional vertebrae at the thoracolumbar junction and establish a technique to differentiate the various subtypes that may be found. A selection of vertebral columns from skeletal remains (n = 35) were evaluated in this study. Vertebrae were taken based on features that are atypical for vertebrae in each relative region. The transitional vertebrae were qualitatively identified based on overlapping thoracic and lumbar features of vertebrae at the thoracolumbar junction. The following general overlapping characteristics were observed: aplasia or hypoplasia of the transverse process, irregular orientation on the superior articular process and atypical mammillary bodies. The results show that the most frequent location of the transitional vertebrae was in the thoracic region (f = 23). The second most frequent location was in the lumbar region (f = 10). In two specimens of the selection (f = 2), an additional 13th thoracic vertebra was present which functioned as a transitional vertebra. This study concluded that one can accurately identify the characteristics of transitional vertebrae at the thoracolumbar junction. In addition, the various subtypes can be differentiated according to the region in the vertebral column the vertebra is located in and the relative number of vertebral segments in the adjacent regions of the vertebral column. This provides a qualitative tool for researchers to differentiate the transitional vertebrae from distinctly different typical thoracic or lumbar vertebrae at the thoracolumbar junction.     

Evolutionary treasures hidden in the West Indies: Comparative osteology and visceral morphology reveals intricate miniaturization in the insular genera Mitophis Hedges,Adalsteinsson, & Branch, 2009 and Tetracheilostoma Jan, 1861 (Leptotyphlopidae: Epictinae: Tetracheilostomina)

The genera Mitophis and Tetracheilostoma comprise two extant lineages of small-sized threadsnakes that exclusively inhabit several islands of the West Indies. Even though leptotyphlopids are known for their extremely reduced size, miniaturization has only been hypothesized to reflect insular dwarfism for the genus Tetracheilostoma. Herein, we aim to describe the comparative osteology and visceral morphology of both genera, investigating and discussing their several internal morphological simplifications and novelties. Our results indicate that these taxa exhibit several autapomorphies mostly concentrated in the dorsoposterior skull elements and maxillae, as well as in their axial skeleton and viscera. These novelties and simplifications are most likely a result of extreme miniaturization driven by the evolutionary constraints or ecological opportunities possibly imposed by the “island rule.” Both Mitophis and Tetracheilostoma distinguish from all other Epictinae in lacking a dentigerous process in the maxillae, by having the prootic fused to the otooccipital, and by the lack (except in comparison to a few Epictia) of a cervical vertebrae intercentrum I. Additionally, Mitophis can be distinguished from other Epictinae by the participation of the unpaired supraoccipital in the dorsal border of the foramen magnum, by the absence of the pleurapophyses in the caudal vertebrae, by a higher number of liver segments, and by the extreme degeneration of the pelvic rudiments. Tetracheilostoma differs from other Epictinae by lacking a distinct supraoccipital, which is fused to the parietal. Thus, our results reinforce that morphological characters are extremely valuable for leptotyphlopid systematics given their extremely conserved external morphology.     

The Key to Understanding the European Miocene Chalcides (Squamata,Scincidae) Comes from Asia: The Lizards of the East Siberian Tagay Locality (Baikal Lake) in Russia

The early middle Miocene (MN 5) lizards from the East Siberian Tagay locality (Baikal Lake, Russia) in Asia are described here. The lizard fauna consists of two clades, Lacertidae and Scincidae. The skink material is allocated to Chalcides. While this taxon was previously reported from Europe, it has rarely been observed in the Neogene record with only jaw fragments and frontal bones described. Its taxonomy was therefore enigmatic. The Tagay material is almost identical to the European fossils of Chalcides from Austria and Hungary, but it also contains the parietal bone. While the material is also similar to the extant Ch. ocellatus, it exhibits several morphological differences. A new species is therefore erected—Chalcides augei sp. nov. These findings further support the connection of the Baikal Lake area with central Europe during the first half of the Miocene. The comparative anatomy of the frontals, parietals and lower jaws was evaluated by micro-CT in selected skink taxa. This comparison highlights several important differences, for example, paired frontals are present in Broadleysaurus (an outgroup taxon), in Acontias and all studied members of Scincidae herein. The character optimization in Mesquite supports fused frontals as being the condition at the basal node of the Ateuchosauridae + Sphenomorphidae + Eugongylidae + Lygosomidae + Egerniidae + Mabuyidae clade. While the parapineal foramen is restricted to the parietal in most taxa studied herein, it is absent (or vestigial) in Acontias and Feylinia. In contrast to all other skinks, this foramen is located on the frontal in Ateuchosaurus chinensis. Anat Rec, 2019. © 2019 American Association for Anatomy Anat Rec, 303:1901–1934, 2020. © 2019 American Association for Anatomy     

The inner ear of Megatherium and the evolution of the vestibular system in sloths

Extant tree sloths are uniquely slow mammals with a very specialized suspensory behavior. To improve our understanding of their peculiar evolution, we investigated the inner ear morphology of one of the largest and most popular fossil ground sloths, Megatherium americanum. We first address the predicted agility of this animal from the scaling of its semicircular canals (SC) relative to body mass, based on recent work that provided evidence that the size of the SC in mammals correlates with body mass and levels of agility. Our analyses predict intermediate levels of agility for Megatherium, contrasting with the extreme slowness of extant sloths. Secondly, we focus on the morphology of the SC at the inner ear scale and investigate the shape and proportions of these structures in Megatherium and in a large diversity of extant xenarthrans represented in our database. Our morphometric analyses demonstrate that the giant ground sloth clearly departs from the SC morphology of both extant sloth genera (Choloepus, Bradypus) and is in some aspects closer to that of armadillos and anteaters. Given the close phylogenetic relationships of Megatherium with the extant genus Choloepus, these results are evidence of substantial homoplasy of the SC anatomy in sloths. This homoplasy most likely corresponds to an outstanding convergent evolution between extant suspensory sloth genera.     

Developmental Failure of Segmentation in a Caudal Vertebra of Apatosaurus (Sauropoda)

A vertebral element assigned to an Apatosaurus cf. ajax from the Late Jurassic Morrison Formation is described. The specimen exhibits an unusual morphology where two vertebrae are nearly seamlessly fused together, including the haemal arch that spans them. This morphology is thought be the result of a developmental abnormality. CT scans of the specimen reveal a thin zone of dorsoventral thickening between the two neural arches consistent with cortical bone. Contrast in internal morphology differentiates the anterior and posterior vertebral bodies with the anterior expressing greater porosity, which increased accommodation for barite‐rich calcite precipitation. No vacuities are observed to suggest the former presence of an intervertebral disk or intervertebral joints: the absence of an intervertebral disc or intervertebral joints is indicative of a condition known as block vertebra. Block vertebrae occur with the loss, or inhibition, of somitocoele mesenchyme early in embyogenesis (i.e., during resegmentation of the somites responsible for the formation of the affected vertebra). The derivatives of somitocoele mesenchyme include the intervertebral disc and joints. Although vertebral paleopathologies are not uncommon in the fossil record, this specimen is the first recognized congenital malformation within Sauropoda. Anat Rec, 297:1262–1269, 2014. © 2014 Wiley Periodicals, Inc.     

Vertebral column regionalisation in Chinook salmon,Oncorhynchus tshawytscha

Teleost vertebral centra are often similar in size and shape, but vertebral‐associated elements, i.e. neural arches, haemal arches and ribs, show regional differences. Here we examine how the presence, absence and specific anatomical and histological characters of vertebral centra‐associated elements can be used to define vertebral column regions in juvenile Chinook salmon (Oncorhynchus tshawytscha). To investigate if the presence of regions within the vertebral column is independent of temperature, animals raised at 8 and 12 °C were studied at 1400 and 1530 degreedays, in the freshwater phase of the life cycle. Anatomy and composition of the skeletal tissues of the vertebral column were analysed using Alizarin red S whole‐mount staining and histological sections. Six regions, termed I–VI, are recognised in the vertebral column of specimens of both temperature groups. Postcranial vertebrae (region I) carry neural arches and parapophyses but lack ribs. Abdominal vertebrae (region II) carry neural arches and ribs that articulate with parapophyses. Elastic‐ and fibrohyaline cartilage and Sharpey's fibres connect the bone of the parapophyses to the bone of the ribs. In the transitional region (III) vertebrae carry neural arches and parapophyses change stepwise into haemal arches. Ribs decrease in size, anterior to posterior. Vestigial ribs remain attached to the haemal arches with Sharpey's fibres. Caudal vertebrae (region IV) carry neural and haemal arches and spines. Basidorsals and basiventrals are small and surrounded by cancellous bone. Preural vertebrae (region V) carry neural and haemal arches with modified neural and haemal spines to support the caudal fin. Ural vertebrae (region VI) carry hypurals and epurals that represent modified haemal and neural arches and spines, respectively. The postcranial and transitional vertebrae and their respective characters are usually recognised, but should be considered as regions within the vertebral column of teleosts because of their distinctive morphological characters. While the number of vertebrae within each region can vary, each of the six regions is recognised in specimens of both temperature groups. This refined identification of regionalisation in the vertebral column of Chinook salmon can help to address evolutionary developmental and functional questions, and to support applied research into this farmed species.     

Farewell to life on land – thoracic strength as a new indicator to determine paleoecology in secondary aquatic mammals

Habitat shifts from land to water have occurred independently in several mammal lineages. However, because we do not know completely about the relationship between skeletal morphology and function, both reliable life reconstructions of each extinct taxon and the timing of those shifts in locomotor strategies are yet to be fully understood. We estimated the strengths of rib cages against vertical compression in 26 extant and four extinct mammal specimens including cetartiodactyls, paenungulates, and carnivorans, representing 11 terrestrial, six semi‐aquatic, and nine obligate aquatic taxa. Our analyses of extant taxa showed that strengths were high among terrestrial/semi‐aquatic mammals, whose rib cages are subjected to vertical compression during the support on land, whereas strengths were low among obligate aquatic mammals, whose rib cages are not subjected to antigravity force in the water. We therefore propose rib strength as a new index to estimate the ability of an animal to be supported on land while being supported by either the forelimbs or thoracic region. According to our analyses of extinct taxa, this ability to be supported on land was rejected for a basal cetacean (Cetartiodactyla: Ambulocetus) and two desmostylians (Paenungulata: Paleoparadoxia and Neoparadoxia). However, this ability was not rejected for one desmostylian species (Desmostylus). Further study of the ribs of extant/extinct semi‐aquatic taxa may help in understanding the ecological shifts in these groups.     

Chromosome painting in Tragulidae facilitates the reconstruction of Ruminantia ancestral karyotype

Although Tragulidae, as the basal family in Ruminantia phylogenetic tree, is the key taxon for understanding the early chromosome evolution of extant ruminants, comparative molecular cytogenetic data on the tragulids are scarce. Here, we present the first genome-wide comparative map of the Java mouse deer (Tragulus javanicus, Tragulidae) revealed by chromosome painting with human and dromedary probes. Together with the published comparative maps of major representative cetartiodactyl species established with the same set of probes, our results allowed us to reconstruct a 2n = 48 Ruminantia ancestral karyotype, which is similar to the cetartiodactyl ancestral karyotype. The karyotype evolution of T. javanicus has involved multiple rearrangements, most of which appear to be apomorphic and have not found in karyotype evolution of pecoran species (i.e., Ruminantia excluding Tragulidae). The rate of chromosome evolution of the mouse deer was rather low—0.4 R/Ma, while the estimated tempo of chromosome changes on the lineages leading from Cetartiodactyla ancestor to Ruminantia and from Ruminantia to Pecora were roughly the same (about 1.2 R/Ma).     

Histology of the Skin of Three Limbless Squamates Dwelling in Mesic and Arid Environments

The skin of limbless squamates has an increased contact with the substrate compared with limbed counterparts. Comparatively, the contact with the substrate is intensified in fossorial species, where the whole circumference of the body interacts with the soil during underground locomotion. Although fossoriality in Squamata, specifically lizards and snakes, has been studied ecologically and morphologically (e.g., osteological changes), not enough detail is yet available regarding changes in organs critical for underground lifestyle such as the skin. Here we used histological and microscopical techniques (scanning electron microscopy and transmission electron microscopy) to uncover the structural detail of the epidermis and dermis in three limbless reptiles, the amphisbaenian Diplometopon zarudnyi, and two snakes, Indotyphlops braminus (Typhlopidae) and Cerastes cerastes (Viperidae). The skin of these taxa shows pronounced morphological diversity, which is likely associated to different environmental and functional demands upon these reptiles. Anat Rec, 299:979–989, 2016. © 2016 Wiley Periodicals, Inc.     

Development and evolution of the notarium in Pterosauria

The notarium is the structure formed by fusion of the dorsal vertebrae which occurred independently in pterosaurs and birds. This ankylosis usually involves two to six elements and in many cases, also includes the last cervical vertebra. Fusion can occur in different degrees, uniting the vertebral centra, the neural spines, the transverse processes, the ventral processes, or a combination of these sites. A detailed assessment of the fusion process of pterosaur dorsal vertebrae is still lacking. Here we identify the fusion sequence of pterosaur notarial elements, demonstrating the order of ossification in vertebral bodies and neural spines based on fossils and extant birds. In both Pterosauria and Aves, the notarium generally develops in a antero-posterior direction, but the actual order of each fusion locus may present slight variations. Based on our data, we were able to identify seven developmental stages in the notarium formation, with broad implications for the prediction of ontogenetic stages for the Pterosauria. In addition, we report the occurrence of a notarium in Ardeadactylus longicollum (Kimmeridgian, Southern Germany), the oldest occurrence of this structure in pterosaurs.