Size and shape heterodonty in the early Permian synapsid Mesenosaurus efremovi.

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.

Dental anatomy and replacement patterns in the early Permian stem amniote, Seymouria.

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.

Neurosensory anatomy of Varanopidae and its implications for early synapsid evolution.

Varanopids are a group of Palaeozoic terrestrial amniotes which represent one of the earliest-diverging groups of synapsids, but their palaeoneurology has gone largely unstudied and recent analyses have challenged their traditional placement within synapsids. We utilized computed tomography (CT) to study the virtual cranial and otic endocasts of six varanopids, including representative taxa of both mycterosaurines and varanodontines. Our results show that the varanopid brain is largely plesiomorphic, being tubular in shape and showing no expansion of the cerebrum or olfactory bulbs, but is distinct in showing highly expanded floccular fossae. The housing of the varanopid bony labyrinth is also distinct, in that the labyrinth is bounded almost entirely by the supraoccipital-opisthotic complex, with the prootic only bordering the ventral portion of the vestibule. The bony labyrinth is surprisingly well-ossified, clearly preserving the elliptical, sub-orthogonal canals, prominent ampullae, and the short, undifferentiated vestibule; this high degree of ossification is similar to that seen in therapsid synapsids and supports the traditional placement of varanopids within Synapsida. The enlarged anterior canal, together with the elliptical, orthogonal canals and enlarged floccular fossa, lend support for the fast head movements indicated by the inferred predatory feeding mode of varanopids. Reconstructed neurosensory anatomy indicates that varanopids may have a much lower-frequency hearing range compared to more derived synapsids, suggesting that, despite gaining some active predatory features, varanopids retain plesiomorphic hearing capabilities. As a whole, our data reveal that the neuroanatomy of pelycosaur-grade synapsids is far more complex than previously anticipated.

Retention of fish-like odontode overgrowth in Permian tetrapod dentition supports outside-in theory of tooth origins.

Teeth are often thought of as structures that line the margins of the mouth; however, tooth-like structures called odontodes are commonly found on the dermal bones of many Palaeozoic vertebrates including early jawless fishes. 'Odontode' is a generalized term for all tooth-like dentine structures that have homologous tissues and development. This definition includes true teeth and the odontodes of early 'fishes', which have been recently examined to gain new insights into the still unresolved origin of teeth. Two leading hypotheses are frequently referenced in this debate: the 'outside-in' hypothesis, which posits that dermal odontodes evolutionarily migrate into the oral cavity, and the 'inside-out' hypothesis, which posits that teeth originated in the oropharyngeal cavity and then moved outwards into the oral cavity. Here, we show that, unlike the well-known one-to-one replacement patterns of marginal dentition, the palatal dentition of the early Permian tetrapods, including the dissorophoid amphibian Cacops and the early reptile Captorhinus, is overgrown by a new layer of bone to which the newest teeth are then attached. This same overgrowth pattern has been well documented in dermal and oral odontodes (i.e. teeth) of early fishes. We propose that this pattern represents the primitive condition for vertebrates and may even predate the origin of jaws. Therefore, this pattern crosses the fish-tetrapod transition, and the retention of this ancestral pattern in the palatal dentition of early terrestrial tetrapods provides strong support for the 'outside-in' hypothesis of tooth origins.

Dimetrodon (Synapsida: Sphenacodontidae) from the cave system at Richards Spur, OK, USA, and a comparison of Early Permian-aged vertebrate paleoassemblages.

The Early Permian Richards Spur locality is unique in preserving a highly diverse faunal assemblage in a cave system, composed of synapsids, reptiles, and anamniotes. However, the presence of Dimetrodon, the most common synapsid of Early Permian localities of the southwestern USA, has never been recorded from the site. Here, we describe for the first time the morphology and histology of a small neural spine with the distinctive figure-8 shape attributable to Dimetrodon. Additionally, histological analysis of previously described sphenacodontid teeth suggests the presence of a derived species of Dimetrodon at the Richards Spur locality. The presence of this derived synapsid, typical of the later occurring Kungurian localities of Texas and Oklahoma, is unexpected at the stratigraphically older Richards Spur locality. The cave system at Richards Spur preserves mainly basal synapsid taxa, including small caseid, varanopid, and sphenacodontid skeletal remains. The presence of a derived species of Dimetrodon suggests not only that this animal was more widespread than previously thought, but that there are different patterns of Early Permian synapsid evolution in different ecological settings.

Embryology of Early Jurassic dinosaur from China with evidence of preserved organic remains.

Fossil dinosaur embryos are surprisingly rare, being almost entirely restricted to Upper Cretaceous strata that record the late stages of non-avian dinosaur evolution. Notable exceptions are the oldest known embryos from the Early Jurassic South African sauropodomorph Massospondylus and Late Jurassic embryos of a theropod from Portugal. The fact that dinosaur embryos are rare and typically enclosed in eggshells limits their availability for tissue and cellular level investigations of development. Consequently, little is known about growth patterns in dinosaur embryos, even though post-hatching ontogeny has been studied in several taxa. Here we report the discovery of an embryonic dinosaur bone bed from the Lower Jurassic of China, the oldest such occurrence in the fossil record. The embryos are similar in geological age to those of Massospondylus and are also assignable to a sauropodomorph dinosaur, probably Lufengosaurus. The preservation of numerous disarticulated skeletal elements and eggshells in this monotaxic bone bed, representing different stages of incubation and therefore derived from different nests, provides opportunities for new investigations of dinosaur embryology in a clade noted for gigantism. For example, comparisons among embryonic femora of different sizes and developmental stages reveal a consistently rapid rate of growth throughout development, possibly indicating that short incubation times were characteristic of sauropodomorphs. In addition, asymmetric radial growth of the femoral shaft and rapid expansion of the fourth trochanter suggest that embryonic muscle activation played an important role in the pre-hatching ontogeny of these dinosaurs. This discovery also provides the oldest evidence of in situ preservation of complex organic remains in a terrestrial vertebrate.

Dental ontogeny in extinct synapsids reveals a complex evolutionary history of the mammalian tooth attachment system.

The mammalian dentition is uniquely characterized by a combination of precise occlusion, permanent adult teeth and a unique tooth attachment system. Unlike the ankylosed teeth in most reptiles, mammal teeth are supported by a ligamentous tissue that suspends each tooth in its socket, providing flexible and compliant tooth attachment that prolongs the life of each tooth and maintains occlusal relationships. Here we investigate dental ontogeny through histological examination of a wide range of extinct synapsid lineages to assess whether the ligamentous tooth attachment system is unique to mammals and to determine how it evolved. This study shows for the first time that the ligamentous tooth attachment system is not unique to crown mammals within Synapsida, having arisen in several non-mammalian therapsid clades as a result of neoteny and progenesis in dental ontogeny. Mammalian tooth attachment is here re-interpreted as a paedomorphic condition relative to the ancestral synapsid form of tooth attachment.

The Permian reptile Opisthodontosaurus carrolli: a model for acrodont tooth replacement and dental ontogeny.

Continuous tooth replacement is common for tetrapods, but some groups of acrodont lepidosaurs have lost the ability to replace their dentition (monophyodonty). Acrodonty, where the tooth attaches to the apex of the jawbone, is an unusual form of tooth attachment that has been associated with the highly autapomorphic condition of monophyodonty. Beyond Lepidosauria, very little is known about the relationship between acrodonty and monophyodonty in other amniotes. We test for this association with a detailed study of the dentition of Opisthodontosaurus, an unusual Early Permian captorhinid eureptile with acrodont dentition. We provide clear evidence, both histological and morphological, that there were regular tooth replacement events in the lower jaw of Opisthodontosaurus, similar to its captorhinid relatives. Thus, our study of the oldest known amniote with an acrodont dentition shows that acrodonty does not inhibit tooth replacement, and that many of the characteristics assigned to lepidosaurian acrodonty are actually highly derived features of lepidosaurs that have resulted secondarily from a lack of tooth replacement. In the context of reptilian dental evolution, we propose the retention of the simple definition of acrodonty, which only pertains to the relative position of the tooth at the apex of the jaw, where the jaw possesses equal lingual and labial walls. This definition of implantation therefore focuses solely on the spatial relationship between the tooth and the jawbone, and separates this relationship from tooth development and replacement.

Eocene lizard from Germany reveals amphisbaenian origins.

Amphisbaenia is a speciose clade of fossorial lizards characterized by a snake-like body and a strongly reinforced skull adapted for head-first burrowing. The evolutionary origins of amphisbaenians are controversial, with molecular data uniting them with lacertids, a clade of Old World terrestrial lizards, whereas morphology supports a grouping with snakes and other limbless squamates. Reports of fossil stem amphisbaenians have been falsified, and no fossils have previously tested these competing phylogenetic hypotheses or shed light on ancestral amphisbaenian ecology. Here we report the discovery of a new lacertid-like lizard from the Eocene Messel locality of Germany that provides the first morphological evidence for lacertid-amphisbaenian monophyly on the basis of a reinforced, akinetic skull roof and braincase, supporting the view that body elongation and limblessness in amphisbaenians and snakes evolved independently. Morphometric analysis of body shape and ecology in squamates indicates that the postcranial anatomy of the new taxon is most consistent with opportunistically burrowing habits, which in combination with cranial reinforcement indicates that head-first burrowing evolved before body elongation and may have been a crucial first step in the evolution of amphisbaenian fossoriality.

Ontogeny reveals function and evolution of the hadrosaurid dinosaur dental battery.

BACKGROUND: Hadrosaurid dinosaurs, dominant Late Cretaceous herbivores, possessed complex dental batteries with up to 300 teeth in each jaw ramus. Despite extensive interest in the adaptive significance of the dental battery, surprisingly little is known about how the battery evolved from the ancestral dinosaurian dentition, or how it functioned in the living organism. We undertook the first comprehensive, tissue-level study of dental ontogeny in hadrosaurids using several intact maxillary and dentary batteries and compared them to sections of other archosaurs and mammals. We used these comparisons to pinpoint shifts in the ancestral reptilian pattern of tooth ontogeny that allowed hadrosaurids to form complex dental batteries. RESULTS: Comparisons of hadrosaurid dental ontogeny with that of other amniotes reveals that the ability to halt normal tooth replacement and functionalize the tooth root into the occlusal surface was key to the evolution of dental batteries. The retention of older generations of teeth was driven by acceleration in the timing and rate of dental tissue formation. The hadrosaurid dental battery is a highly modified form of the typical dinosaurian gomphosis with a unique tooth-to-tooth attachment that permitted constant and perfectly timed tooth eruption along the whole battery. CONCLUSIONS: We demonstrate that each battery was a highly dynamic, integrated matrix of living replacement and, remarkably, dead grinding teeth connected by a network of ligaments that permitted fine scale flexibility within the battery. The hadrosaurid dental battery, the most complex in vertebrate evolution, conforms to a surprisingly simple evolutionary model in which ancestral reptilian tissue types were redeployed in a unique manner. The hadrosaurid dental battery thus allows us to follow in great detail the development and extended life history of a particularly complex food processing system, providing novel insights into how tooth development can be altered to produce complex dentitions, the likes of which do not exist in any living vertebrate.

Mineralized periodontia in extinct relatives of mammals shed light on the evolutionary history of mineral homeostasis in periodontal tissue maintenance.

AIM: Dental ankylosis is a rare pathological condition in mammals, however, it is prevalent in their extinct relatives, the stem mammals. This study seeks to compare the mineralized state of the periodontal attachment apparatus between stem and crown mammals and discuss its implications for the evolution of non-mineralized periodontal attachment in crown mammals, including humans. MATERIALS AND METHODS: Thin sections of a fossil mammal and three stem mammals were compared to reconstruct periodontal tissue development across distantly related lineages. RESULTS: Comparisons revealed that the extinct relatives of mammals possessed the same periodontal tissues as those in mammals, albeit in different arrangements. The ankylotic condition in stem mammals was achieved through extensive alveolar bone deposition, which eventually contacted the root cementum, thus forming a calcified periodontal ligament. CONCLUSIONS: Dental ankylosis was part of the normal development of the stem mammal periodontium for millions of years prior to the evolution of a permanent gomphosis in mammals. Mammals may have evolved a permanent gomphosis by delaying the processes that produced dental ankylosis in stem mammals. Pathological ankylosis may represent a reversion to the ancestral condition, which now only forms via advanced ageing and pathology.

The oldest parareptile and the early diversification of reptiles.

Amniotes, tetrapods that evolved the cleidoic egg and thus independence from aquatic larval stages, appeared ca 314 Ma during the Coal Age. The rapid diversification of amniotes and other tetrapods over the course of the Late Carboniferous period was recently attributed to the fragmentation of coal-swamp rainforests ca 307 Ma. However, the amniote fossil record during the Carboniferous is relatively sparse, with ca 33% of the diversity represented by single specimens for each species. We describe here a new species of reptilian amniote that was collected from uppermost Carboniferous rocks of Prince Edward Island, Canada. Erpetonyx arsenaultorum gen. et sp. nov. is a new parareptile distinguished by 29 presacral vertebrae and autapomorphies of the carpus. Phylogenetic analyses of parareptiles reveal E. arsenaultorum as the closest relative of bolosaurids. Stratigraphic calibration of our results indicates that parareptiles began their evolutionary radiation before the close of the Carboniferous Period, and that the diversity of end-Carboniferous reptiles is 80% greater than suggested by previous work. Latest Carboniferous reptiles were still half as diverse as synapsid amniotes, a disparity that may be attributable to preservational biases, to collecting biases, to the origin of herbivory in tetrapods or any combination of these factors.

Third-harmonic generation microscopy reveals dental anatomy in ancient fossils.

Fossil teeth are primary tools in the study of vertebrate evolution, but standard imaging modalities have not been capable of providing high-quality images in dentin, the main component of teeth, owing to small refractive index differences in the fossilized dentin. Our first attempt to use third-harmonic generation (THG) microscopy in fossil teeth has yielded significant submicrometer level anatomy, with an unexpectedly strong signal contrasting fossilized tubules from the surrounding dentin. Comparison between fossilized and extant teeth of crocodilians reveals a consistent evolutionary signature through time, indicating the great significance of THG microscopy in the evolutionary studies of dental anatomy in fossil teeth.

Oldest known dinosaurian nesting site and reproductive biology of the Early Jurassic sauropodomorph Massospondylus.

The extensive Early Jurassic continental strata of southern Africa have yielded an exceptional record of dinosaurs that includes scores of partial to complete skeletons of the sauropodomorph Massospondylus, ranging from embryos to large adults. In 1976 an incomplete egg clutch including in ovo embryos of this dinosaur, the oldest known example in the fossil record, was collected from a road-cut talus, but its exact provenance was uncertain. An excavation program at the site started in 2006 has yielded multiple in situ egg clutches, documenting the oldest known dinosaurian nesting site, predating other similar sites by more than 100 million years. The presence of numerous clutches of eggs, some of which contain embryonic remains, in at least four distinct horizons within a small area, provides the earliest known evidence of complex reproductive behavior including site fidelity and colonial nesting in a terrestrial vertebrate. Thus, fossil and sedimentological evidence from this nesting site provides empirical data on reproductive strategies in early dinosaurs. A temporally calibrated optimization of dinosaurian reproductive biology not only demonstrates the primary significance of the Massospondylus nesting site, but also provides additional insights into the initial stages of the evolutionary history of dinosaurs, including evidence that deposition of eggs in a tightly organized single layer in a nest evolved independently from brooding.

First record of plicidentine in Synapsida and patterns of tooth root shape change in Early Permian sphenacodontians.

Recent histological studies have revealed a diversity of dental features in Permo-Carboniferous tetrapods. Here, we report on the occurrence of plicidentine (infolded dentine around the base of the tooth root) in Sphenacodontia, the first such documentation in Synapsida, the clade that includes mammals. Five taxa were examined histologically, Ianthodon schultzei, Sphenacodon ferocior, Dimetrodon limbatus, Dimetrodon grandis, and Secodontosaurus obtusidens. The tooth roots of Ianthodon possess multiple folds, which is generally viewed as the primitive condition for amniotes. Sphenacodon and D. limbatus have distinctive "four-leaf clover"-shaped roots in cross section, whereas Secodontosaurus has an elongate square shape with only subtle folding. The most derived and largest taxon examined in this study, D. grandis, has rounded roots in cross section and therefore no plicidentine. This pattern of a loss of plicidentine in sphenacodontids supports previous functional hypotheses of plicidentine, where teeth with shallow roots require folds to increase the area of attachment to the tooth-bearing element, whereas teeth with long roots do not. This pattern may also reflect differences in diet between co-occurring sphenacodontids as well as changes in feeding niche through time, specifically in the apex predator Dimetrodon.

A stem batrachian from the Early Permian of Texas and the origin of frogs and salamanders.

The origin of extant amphibians (Lissamphibia: frogs, salamanders and caecilians) is one of the most controversial questions in vertebrate evolution, owing to large morphological and temporal gaps in the fossil record. Current discussions focus on three competing hypotheses: a monophyletic origin within either Temnospondyli or Lepospondyli, or a polyphyletic origin with frogs and salamanders arising among temnospondyls and caecilians among the lepospondyls. Recent molecular analyses are also controversial, with estimations for the batrachian (frog-salamander) divergence significantly older than the palaeontological evidence supports. Here we report the discovery of an amphibamid temnospondyl from the Early Permian of Texas that bridges the gap between other Palaeozoic amphibians and the earliest known salientians and caudatans from the Mesozoic. The presence of a mosaic of salientian and caudatan characters in this small fossil makes it a key taxon close to the batrachian (frog and salamander) divergence. Phylogenetic analysis suggests that the batrachian divergence occurred in the Middle Permian, rather than the late Carboniferous as recently estimated using molecular clocks, but the divergence with caecilians corresponds to the deep split between temnospondyls and lepospondyls, which is congruent with the molecular estimates.

Exceptionally rapid tooth development and ontogenetic changes in the feeding apparatus of the Komodo dragon.

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.

The age and growth information of a ctenoid scale fossil from the Upper Cretaceous Nenjiang Formation in Songliao Basin, China.

The study of morphological characteristics and growth information in fish scales is a crucial component of modern fishery biological research, while it has been less studied in fossil materials. This paper presents a detailed morphological description and growth analysis of a fossil ctenoid scale obtained from the Upper Cretaceous Campanian lacustrine deposits in northeastern China. The morphological features of this fossil scale are well-preserved and consistent with the structures found in ctenoid scales of extant fish species and display prominent ring ornamentation radiating outward from the central focus, with grooves intersecting the rings. A comparative analysis of the morphological characteristics between the fossil ctenoid scale and those well-studied extant fish Mugilidae allows us to explore the applicability of modern fishery biological research methods to the field of fossil scales. The scale length, scale width, the vertical distance from the focus to the apex of the scale, and the total number of radii have been measured. The age of the fish that possessed this ctenoid scale has been estimated by carefully counting the annuli, suggesting an age equal to or more than seven years. The distribution of growth rings on the scale potentially reflects the warm paleoclimatic condition and fish-friendly paleoenvironment prevalent during that period. This paper, moreover, serves as a notable application of fishery biological methods in the examination of fossil materials.

Paleozoic cave system preserves oldest-known evidence of amniote skin.

The richest and most diverse assemblage of early terrestrial tetrapods is preserved within the infilled cave system of Richards Spur, Oklahoma (289-286 Mya1). Some of the oldest-known terrestrial amniotes2,3 are exquisitely preserved here because of early impregnation and encasement of organic material by oil-seep hydrocarbons within rapidly deposited clay-rich cave sediments under toxic anoxic conditions.4 This phenomenon has also afforded the preservation of exceedingly rare integumentary soft tissues, reported here, providing critical first evidence into the anatomical changes marking the transition from the aquatic and semiaquatic lifestyles of anamniotes to the fully terrestrial lifestyles of early amniotes. This is the first record of a skin-cast fossil (3D carbonization of the skin proper) from the Paleozoic Era and the earliest known occurrence of epidermal integumentary structures. We also report on several compression fossils (carbonized skin impressions), all demonstrating similar external morphologies to extant crocodiles. A variety of previously unknown ossifications, as well as what are likely palpebral ossifications of the deeper dermis layer of the skin, are also documented. These fossils also serve as invaluable references for paleontological reconstructions. Chromatographic analysis of extractable hydrocarbons from bone and cave samples indicates that the source rock is the Devonian age Woodford Shale. Hydrocarbons derived from ancient marine organisms interacting with geologically younger terrestrial vertebrates have therefore resulted in the oldest-known preservation of amniote skin proper.