Tea Maho

Costal aspiration breathing was an evolutionary innovation that was fundamental to the conquest of the terrestrial realm by amniotes (mammals, reptiles, birds and their common ancestor)^1,2,3,4,5. Extant amniotes use an integrated thoracic skeleton for costal-muscle-generated inhalation and exhalation, differing substantially from their anamniote relatives, which possess more passive cutaneous and buccal pumping forms of ventilation. This difference extends into the Palaeozoic era, but the evolutionary transformation between these modes of breathing is undocumented and largely unclear^6,7,8,9,10 in the absence of soft tissue fossils. Here we present the mummified early Permian reptile Captorhinus, which includes a covering of three-dimensional skin, native protein remnants and a complete shoulder girdle and ribcage with preserved cartilages. These are the oldest-known preserved cartilages and protein remnants in a terrestrial vertebrate. High-resolution neutron computed tomography and histology data reveal previously undescribed structures, including the cartilaginous sternum, sternal ribs, rib extensions and epicoracoids. Our skeletal reconstruction of this ancient reptile reveals the precise relationships between the ribcage and the shoulder girdle, and their pivotal role in the evolution of terrestrial breathing and locomotor regimes^11,12. This finding substantially changes expectations of soft tissue preservation in deep time to reveal the potential ancestral amniote breathing mechanism and its impact on terrestrial vertebrate evolution.

Paleozoic synapsids represent the first chapter in the evolution of this large clade that includes mammals. These fascinating terrestrial vertebrates were the first amniotes to successfully adapt to a wide range of feeding strategies, reflected by their varied dental morphologies. Evolution of the marginal dentition on the mammalian side of amniotes is characterized by strong, size and shape heterodonty, with the late Permian therapsids showing heterodonty with the presence of incisiform, caniniform, and multicuspid molariform dentition. Rarity of available specimens has previously prevented detailed studies of dental anatomy and evolution in the initial chapter of synapsid evolution, when synapsids were able to evolve dentition for insectivory, herbivory, and carnivory. Numerous teeth, jaw elements, and skulls of the hypercarnivorous varanopid Mesenosaurus efremovi have been recently discovered in the cave systems near Richards Spur, Oklahoma, permitting the first detailed investigation of the dental anatomy of a Paleozoic tetrapod using multiple approaches, including morphometric and histological analyses. As a distant stem mammal, Mesenosaurus is the first member of this large and successful clade to exhibit a type of dental heterodonty that combines size and morphological (shape) variation of the tooth crowns. Here we present the first evidence of functional differentiation in the dentition of this early synapsid, with three distinct dental regions having diverse morphologies and functions. The quality and quantity of preserved materials has allowed us to identify the orientation and curvature of the carinae (cutting edges), and the variation and distribution of the ziphodonty (serrations) along the carinae. The shape-related heterodonty seen in this taxon may have contributed to this taxon's ability to be a successful mid-sized predator in the taxonomically diverse community of early Permian carnivores, but may have also extended the ecological resilience of this clade of mid-sized predators across major faunal and environmental transitions.

Large isolated skeletal elements, including those of sphenacodontid and ophiacodontid synapsids from the upland cave systems of the Richards Spur locality, Oklahoma, are described. Multiple forms of visual representation, including coquille and stipple drawings, are used to document and examine the isolated elements. A fragmentary anterior portion of a dentary has the sphenacodontid four-leaf clover plicidentine attachment within the tooth roots, but the teeth are all of uniform size, and the symphyseal area is relatively slender and does not curve upward, suggesting that the specimen may not belong to any known member of the clade. A larger humerus with only the distal end preserved and a complete astragalus have distinct characteristics which are attributable to the sphenacodontid Dimetrodon Cope, 1878. A second, smaller humerus was identified to belong to Ophiacodon Marsh, 1878, cf. O. navajovicus and represents the first record of an ophiacodontid at Richards Spur. Finally, two large sphenacodontid interclavicles were discovered, with one having unusual growths representing a pathological condition. Typically, large amniotes are quite rare in this early Permian upland ecosystem, but the discovery of the new material shows that large synapsids are present at Richards Spur.

Dental developmental and replacement patterns in extinct amniotes have attracted a lot of attention. Notable among these are Paleozoic predatory synapsids, but also Mesozoic theropod dinosaurs, well known for having true ziphodonty, strongly serrated carinae with dentine cores within an enamel cap. The Komodo dragon, Varanus komodoensis, is the only extant terrestrial vertebrate to exhibit true ziphodonty, making it an ideal model organism for gaining new insights into the life history and feeding behaviours of theropod dinosaurs and early synapsids. We undertook a comparative dental histological analysis of this extant apex predator in combination with computed tomography of intact skulls. This study allowed us to reconstruct the dental morphology, ontogeny, and replacement patterns in the largest living lizard with known feeding behaviour, and apply our findings to extinct taxa where the behaviour is largely unknown. We discovered through computed tomography that V. komodoensis maintains up to five replacement teeth per tooth position, while histological analysis showed an exceptionally rapid formation of new teeth, every 40 days. Additionally, a dramatic ontogenetic shift in the dental morphology of V. komodoensis was also discovered, likely related to changes in feeding preferences and habitat. The juveniles have fewer dental specializations, lack true ziphodonty, are arboreal and feed mostly on insects, whereas the adults have strongly developed ziphodonty and are terrestrial apex predators with defleshing feeding behaviour. In addition, we found evidence that the ziphodont teeth of V. komodoensis have true ampullae (interdental folds for strengthening the serrations), similar to those found only in theropod dinosaurs. Comparisons with other species of Varanus and successive outgroup taxa reveal a complex pattern of dental features and adaptations, including the evolution of snake-like tongue flicking used for foraging for prey. However, only the Komodo dragon exhibits this remarkable set of dental innovations and specializations among squamates.

Large theropod dinosaurs are often reconstructed with their marginal dentition exposed because of the enormous size of their teeth and their phylogenetic association to crocodylians. We tested this hypothesis using a multiproxy approach. Regressions of skull length and tooth size for a range of theropods and extant varanid lizards confirm that complete coverage of theropod dinosaur teeth with extraoral tissues (gingiva and labial scales) is both plausible and consistent with patterns observed in living ziphodont amniotes. Analyses of dental histology from crocodylians and theropod dinosaurs, including Tyrannosaurus rex , further indicate that the most likely condition was complete coverage of the marginal dentition with extraoral tissue when the mouth was closed. This changes our perceptions about the appearance and oral configuration of these iconic predators and has broad implications for our interpretations of other terrestrial animals with large teeth.

An articulated pelvic region and additional isolated material of Varanops brevirostris, which are indistinguishable from those of the generotype from the Cacops bonebed, demonstrate the presence of this large varanopid at the Richards Spur locality. The articulated specimen includes lumbar, sacral, and anterior caudal vertebrae, partial pelvis, femur, and proximal part of tibia, confirming the autapomorphies previously suggested for this species. These include the presence of distinct blade-like shapes of the neural spines in the sacral region, the presence of deeply excavated pubis, and the presence of a distinct transverse ridge on the ventral surface of the femur distal to the intertrochanteric fossa. It has also been found that the transverse ridges and grooves become larger during ontogeny since the juvenile specimen did not exhibit a well-developed ridge. Histological analysis of isolated limb bones and neutron computed tomography (nCT) of the articulated specimen indicate that the latter likely belonged to an adult individual. This is in contrast to the other varanopid at Richards Spur, the significantly smaller, more gracile predator Mesenosaurus efremovi, which also shows the presence of growth lines and the external fundamental system with an estimated minimum age of fourteen.

The initial stages of diapsid evolution, the clade that includes extant reptiles and the majority of extinct reptilian taxa, is surprisingly poorly known. Notwithstanding the hypothesis that varanopids are diapsids rather than synapsids, there are only four araeoscelidians and one neodiapsid present in the late Carboniferous and early Permian. Here we describe the fragmentary remains of a very unusual new amniote from the famous cave deposits near Richards Spur, Oklahoma, that we recognize as a diapsid reptile, readily distinguishable from all other early amniotes by the unique dentition and lower jaw anatomy. The teeth have an unusual reeding pattern on the crown (long parallel ridges with rounded surfaces), with some teeth posteriorly tilted and strongly recurved, while a ventral protuberance forms the anterior terminus of the dentary. Overall, the lower jaw is unusually slender with a flattened ventral surface formed by the dentary and splenial anteriorly and the angular in the mid-region. The presence of a very slender triradiate jugal revealed through computed tomography confirms the existence of a large lower temporal fenestra, while the medial edge of the maxilla and the anatomy of the palatine confirm the presence of a large suborbital fenestra. Computed tomography of this new taxon reveals maxillary innervation that is characteristically reptile, not synapsid. Although no other definitively identifiable skull roof elements exist, the suborbital fenestra borders preserved on the palatine and maxilla supports the hypothesis that this is a diapsid reptile. Interestingly, the right dentary shows evidence of pathology, a rarely reported occurrence in Paleozoic amniotes, with several empty tooth sockets filled by bone. This small predator with delicate subthecodont implanted dentition provides strong evidence that diapsid reptiles were already diversifying rapidly in the early Permian, but likely were relatively rare members of terrestrial vertebrate assemblages.

The exquisite preservation of maxillary and mandibular fragments of Seymouria has allowed us to examine for the first time in detail the dental anatomy and patterns of development in this stem amniote. The results obtained through histological examination show that Seymouria has pleurodont implantation with ankylosis of the tooth to the labial side of the jawbone. The dentary and maxillary teeth exhibit similar dental characteristics, such as the attachment bone (alveolar bone) and cementum rising above the jawbone on the base of the tooth, and smooth carinae extending lingually toward the tooth apex. Additionally, the clear presence of plicidentine, infolding of dentine into the pulp cavity, was found within the tooth root extending into the tooth crown. Lastly, the tooth replacement pattern is alternating, illustrating that Seymouria retains the classic primitive condition for tetrapods, a pattern that is continued in amniotes. Our results provide an important basis for comparison with other stem amniotes and with amniotes.