The bony labyrinth of Platecarpus (Squamata: Mosasauria) and aquatic adaptations in squamate reptiles

Mosasaurs were among the last marine reptiles that lived before the Cretacesous–Paleogene extinction. Little is known about the sensory evolution of mosasaurs in relation to their aquatic lifestyle. In this study, the braincase of Platecarpus was CT-scanned and virtual models were constructed showing the bony labyrinth — or the inner ear — a sensory apparatus for balance and hearing. The virtual inner ear consists of the semicircular canals, vestibule, and cochlea. Compared with extant squamates, Platecarpus resembles sea snakes in having a small vestibule with a flat dorsal surface, but it differs from non-mosasaurian squamates in having rounded semicircular canals. Phylogenetic linear regression analysis supports a linear relationship, independent from phylogeny, between the length of the three semicircular canals and the length of the skull. The semicircular canals of Platecarpus are shorter than predicted, but the fossil data fell within the 95% prediction interval calculated from the extant data and the skull length of Platecarpus. Although size reduction of the bony labyrinth has been associated with aquatic adaptions in mammals, our results suggest that in squamates, semicircular canal size is related to skull size rather than habitat preference.     

Intraspecific variation in the domestic cat bony labyrinth revealed by different measurement techniques

The knowledge of intraspecific variation is important to make assumptions on an interspecific level. To study intraspecific variation in the bony labyrinth morphology of the domestic cat, eleven specimens of Felis silvestris catus and two additional subspecies (F. s. lybica, F. s. ornata) were investigated. The sample comprises skulls of adult males and females, as well as juvenile cats. Each bony labyrinth endocast was virtually reconstructed based on µCT scans. To estimate the radius of curvature of each inner ear semicircular canal, three different approaches were tested. The comparison of the different methods resulted in different absolute values for the measured radii. The assumed best structure to precisely characterize the size of a semicircular canal is the inner perimeter. Within the tested sample, the anterior semicircular canal is always the largest, while the posterior semicircular canal is the second largest and the lateral semicircular canal the smallest in most cases. The coefficient of variation lies below 10% for all bony labyrinth measurements within the sample. The inner perimeter values of each semicircular canal are similar within all investigated specimens, even though the skull length of adult cats is twice as long as that of juvenile cats. Thus, inner ear biometry of the domestic cat seems stable throughout growth series and can therefore be used for systematic and ecological studies and the inclusion of juvenile individuals is reasonable. It is noteworthy that the inner perimeter values of the semicircular canals do not vary as much as the values of the angles spanned between the three canals within the sample. The inner ear within the cat skull is oriented about 25° to 31° to the palate (angle between the plane anchored to the lateral semicircular canals (SC) and the plane anchored to the palate). The cochlea coils between 3.00 and 3.25 turns in the investigated sample.     

A Systematic Study on Tooth Enamel Microstructures of Lambdopsalis bulla (Multituberculate,Mammalia) - Implications for Multituberculate Biology and Phylogeny

Tooth enamel microstructure is a reliable and widely used indicator of dietary interpretations and data for phylogenetic reconstruction, if all levels of variability are investigated. It is usually difficult to have a thorough examination at all levels of enamel structures for any mammals, especially for the early mammals, which are commonly represented by sparse specimens. Because of the random preservation of specimens, enamel microstructures from different teeth in various species are often compared. There are few examples that convincingly show intraspecific variation of tooth enamel microstructure in full dentition of a species, including multituberculates. Here we present a systematic survey of tooth enamel microstructures of Lambdopsalis bulla, a taeniolabidoid multituberculate from the Late Paleocene Nomogen Formation, Inner Mongolia. We examined enamel structures at all hierarchical levels. The samples are treated differently in section orientations and acid preparation and examined using different imaging methods. The results show that, except for preparation artifacts, the crystallites, enamel types, Schmelzmuster and dentition types of Lambdopsalis are relatively consistent in all permanent teeth, but the prism type, including the prism shape, size and density, may vary in different portions of a single tooth or among different teeth of an individual animal. The most common Schmelzmuster of the permanent teeth in Lambdopsalis is a combination of radial enamel in the inner and middle layers, aprismatic enamel in the outer layer, and irregular decussations in tooth crown area with great curvature. The prism seam is another comparably stable characteristic that may be a useful feature for multituberculate taxonomy. The systematic documentation of enamel structures in Lambdopsalis may be generalized for the enamel microstructure study, and thus for taxonomy and phylogenetic reconstruction, of multituberculates and even informative for the enamel study of other early mammals.     

An X-radiographic and SEM study of the osseous inner ear of multituberculates and monotremes (Mammalia): implications for mammalian phylogeny and evolution of hearing

Multituberculate petrosals with well-preserved, three-dimensional internal anatomy from the Late Cretaceous/early Paleocene Bug Creek Anthills, Montana, U.S.A., are described from X-radiographic and SEM images, as well as from conventional visual observations, and are compared with the anatomy of the osseous inner ear in monotremes and in primitive non-therian and therian mammals. Results of this study indicate that: (1) the cochlea of at least some multituberculates retained a lagena, previously known only in monotremes among mammals; (2) an enlarged vestibule evolved in several lineages of multituberculates independently, and hence is not a synapomorphy of the order; (3) the cochlear canal lacks osseous laminae in support of the short, wide basilar membrane, which was probably inefficient in responding to high-frequency airborne vibrations; and (4) consequently, bone-conducted hearing in some multituberculate species may have been important in interpretation of their surroundings. Comparisons with the inner ear of monotremes and primitive therians indicate that curvature of the cochlea and cribriform plates for passage of vestibulocochlear nerve branches through the petrosal are unlikely homologues between monotremes and therians. From non-therian to therian mammals, there is a distinct morphological gap in the inner ear transition, characterized by acquisition of a number of neomorphs in the therian inner ear; an intermediate stage has yet to be discovered.     

The Evolution of Bat Vestibular Systems in the Face of Potential Antagonistic Selection Pressures for Flight and Echolocation

The vestibular system maintains the body’s sense of balance and, therefore, was probably subject to strong selection during evolutionary transitions in locomotion. Among mammals, bats possess unique traits that place unusual demands on their vestibular systems. First, bats are capable of powered flight, which in birds is associated with enlarged semicircular canals. Second, many bats have enlarged cochleae associated with echolocation, and both cochleae and semicircular canals share a space within the petrosal bone. To determine how bat vestibular systems have evolved in the face of these pressures, we used micro-CT scans to compare canal morphology across species with contrasting flight and echolocation capabilities. We found no increase in canal radius in bats associated with the acquisition of powered flight, but canal radius did correlate with body mass in bat species from the suborder Yangochiroptera, and also in non-echolocating Old World fruit bats from the suborder Yinpterochiroptera. No such trend was seen in members of the Yinpterochiroptera that use laryngeal echolocation, although canal radius was associated with wing-tip roundedness in this group. We also found that the vestibular system scaled with cochlea size, although the relationship differed in species that use constant frequency echolocation. Across all bats, the shape of the anterior and lateral canals was associated with large cochlea size and small body size respectively, suggesting differential spatial constraints on each canal depending on its orientation within the skull. Thus in many echolocating bats, it seems that the combination of small body size and enlarged cochlea together act as a principal force on the vestibular system. The two main groups of echolocating bats displayed different canal morphologies, in terms of size and shape in relation to body mass and cochlear size, thus suggesting independent evolutionary pathways and offering tentative support for multiple acquisitions of echolocation.     

Digital Reconstruction of the Otic Region and Inner Ear of the Non-Mammalian Cynodont Brasilitherium riograndensis (Late Triassic, Brazil) and Its Relevance to the Evolution of the Mammalian Ear

The external anatomy of the petrosal, the bony labyrinth of the inner ear, and the stapes of Brasilitherium riograndensis (specimen UFRGS-PV-1043-T) were investigated by digital 3D reconstructions based on μCT scan images. Brasilitherium is the most basal taxon bearing a distinct promontorium, although less inflated than that of Morganucodon and still lacking a flat medial facet. A bony wall formed by the petrosal separates the cochlear canal and the vestibule from the brain cavity, with an internal acoustic meatus bearing distinct foramina for the facial nerve (VII) and vestibulocochlear nerve (VIII). The semicircular canals are irregular in shape, the anterior canal being the largest and the lateral one the smallest. Brasilitherium has an elongated but straight cochlear canal. The stapes resembles the morphology of derived non-mammaliaform cynodonts, such as Probainognathus and Pachygenelus, and differs from Thrinaxodon. By the allometric relationship of the cochlear canal and the estimated body mass, Brasilitherium can be grouped with Yunnanodon and Morganucodon in a regression line, which is below the line of mammals and above the line of non-avian reptiles. Brasilitherium fits in a sequence of gradual elongation of the cochlear canal associated with the enhancement in the capacity to hear higher frequencies. Among the constraints that might have triggered these transformations in small, insectivorous, and possibly nocturnal Mesozoic cynodont taxa is the improvement of detecting acoustically active insects.     

Preliminary observations on the morphology of the inner ear of the fetal narwhal (Monodon monoceros)

The inner ear of the narwhal (Monodon monoceros) was investigated on the basis of a complete series of microslides of an early fetus. In this well‐preserved specimen, the cochlea is about twice as large as the vestibular apparatus. The latter exhibits a high degree of specialization. Whereas the utriculus and sacculus are of normal size in comparison with those of other mammals, the semicircular canals are miniaturized but not obliterated, as in the case of some adult toothed whales. In comparison with other mammals, there is relatively little perilymphatic space around the vestibular organ, but the inner radius of the lymphatic ducts comes close to that of the adult human. Although there are some hints that the sensitivity of the vestibular system may be high in this species, detailed analysis of the adult narwhal ear is needed to confirm this inference.     

Restriction of Wnt signaling in the dorsal otocyst determines semicircular canal formation in the mouse embryo

The mouse inner ear develops from a simple epithelial pouch, the otocyst, with the dorsal and ventral portions giving rise to the vestibule and cochlea, respectively. The otocyst undergoes a morphological change to generate flattened saclike structures, known as outpocketings, in the dorsal and lateral regions. The semicircular canals of the vestibule form from the periphery of the outpocketings, with the central region (the fusion plate) undergoing de-epithelialization and disappearing. However, little is known of the mechanism that orchestrates formation of the semicircular canals. We now show that the area of canonical Wnt signaling changes dynamically in the dorsal otocyst during its morphogenesis. The genes for several Wnt ligands were found to be expressed in the dorsal otocyst according to specific patterns, whereas those for secreted inhibitors of Wnt ligands were expressed exclusively in the ventral otocyst. With the use of whole-embryo culture in combination with potent modulators of canonical Wnt signaling, we found that forced persistence of such signaling resulted in impaired formation both of the lateral outpocketing and of the fusion plates of the dorsal outpocketing. Canonical Wnt signaling was found to suppress Netrin1 expression and to preserve the integrity of the outpocketing epithelium. In addition, inhibition of canonical Wnt signaling reduced the size of the otocyst, likely through suppression of cell proliferation and promotion of apoptosis. Our stage-specific functional analysis suggests that strict regulation of canonical Wnt signaling in the dorsal otocyst orchestrates the process of semicircular canal formation.     

Ear ossicle morphology of the Jurassic euharamiyidan Arboroharamiya and evolution of mammalian middle ear

The middle ear bones of Mesozoic mammals are rarely preserved as fossils and the morphology of these ossicles in the earliest mammals remains poorly known. Here, we report the stapes and incus of the euharamiyidan Arboroharamiya from the lower Upper Jurassic (～160 Ma) of northern China, which represent the earliest known mammalian middle ear ossicles. Both bones are miniscule in relation to those in non‐mammalian cynodonts. The skull length/stapedial footplate diameter ratio is estimated as 51.74 and the stapes length as the percentage of the skull length is 4%; both numbers fall into the stapes size ranges of mammals. The stapes is “rod‐like” and has a large stapedial foramen. It is unique among mammaliaforms in having a distinct posterior process that is interpreted as for insertion of the stapedius muscle and homologized to the ossified proximal (stapedial) end of the interhyal, on which the stapedius muscle attached. The incus differs from the quadrate of non‐mammalian cynodonts such as morganucodontids in having small size and a slim short process. Along with lack of the postdentary trough and Meckelian groove on the medial surface of the dentary, the ossicles suggest development of the definitive mammalian middle ear (DMME) in Arboroharamiya. Among various higher‐level phylogenetic hypotheses of mammals, the one we preferred places “haramiyidans” within Mammalia. Given this phylogeny, development of the DMME took place once in the allotherian clade containing euharamiyidans and multituberculates, probably independent to those of monotremes and therians. Thus, the DMME has evolved at least three times independently in mammals. Alternative hypothesis that placed “haramiyidans” outside of Mammalia would require independent acquisition of the DMME in multituberculates and euharamiyidans as well as parallel evolution of numerous derived similarities in the dentition, occlusion pattern, mandibles, cranium, and postcranium between the two groups and between “haramiyidans” and other mammals. J. Morphol. 279:441–457, 2018. © 2016 Wiley Periodicals, Inc.     

Tsukushi is essential for the formation of the posterior semicircular canal that detects gait performance

Tsukushi is a small, leucine-rich repeat proteoglycan that interacts with and regulates essential cellular signaling cascades in the chick retina and murine subventricular zone, hippocampus, dermal hair follicles, and the cochlea. However, its function in the vestibules of the inner ear remains unknown. Here, we investigated the function of Tsukushi in the vestibules and found that Tsukushi deficiency in mice resulted in defects in posterior semicircular canal formation in the vestibules, but did not lead to vestibular hair cell loss. Furthermore, Tsukushi accumulated in the non-prosensory and prosensory regions during the embryonic and postnatal developmental stages. The downregulation of Tsukushi altered the expression of key genes driving vestibule differentiation in the non-prosensory regions. Our results indicate that Tsukushi interacts with Wnt2b, bone morphogenetic protein 4, fibroblast growth factor 10, and netrin 1, thereby controlling semicircular canal formation. Therefore, Tsukushi may be an essential component of the molecular pathways regulating vestibular development.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12079-021-00627-1.     

Functional morphological adaptations of the bony labyrinth in marsupials (Mammalia,Theria)

Diprotodontia represents the largest and ecologically most distinct order of marsupials occurring in Australasian being highly divers in size, locomotion, habitat preferences, feeding, and activity pattern. The spatial orientation in the habitat and therefore the three‐dimensional space is detected by the vestibular system of the inner ear, more precisely by the three semicircular canals. In this study, we investigated the bony labyrinth of diprotodontian and selected non‐diprotodontian marsupial mammals of almost all genera with noninvasive micro‐CT scanning and 3D‐reconstructions. In principal component analyses, the subterranean taxon can be separated from gliding and saltatorial taxa, whereas arboreal species can be separated from saltatorial specimens. The highest PCA loadings of this functional distinction are clearly found in the diameter of the semicircular canals, whereas the overall shape (height, width, length) of the semicircular canals is less important. Additionally, the investigated arboreal and fossorial species of South America are nested in the morphospace of the Australasian taxa. Even if a phylogenetic signal in the anatomy of the bony labyrinth cannot be excluded entirely, the main functional morphological signal of the vestibular system is found in the diameter of the semicircular canals. With the large dataset of extant marsupial mammals analysed here, the locomotion mode of extinct taxa can be inferred in future studies independent of any evidence of postcranial material.     

Convergence vs. Specialization in the ear region of moles (mammalia)

We investigated if and how the inner ear region undergoes similar adaptations in small, fossorial, insectivoran‐grade mammals, and found a variety of inner ear phenotypes. In our sample, afrotherian moles (Chrysochloridae) and the marsupial Notoryctes differ from most other burrowing mammals in their relatively short radii of semicircular canal curvature; chrysochlorids and fossorial talpids share a relatively long interampullar width. Chrysochlorids are unique in showing a highly coiled cochlea with nearly four turns. Extensive cochlear coiling may reflect their greater ecological dependence on low frequency auditory cues compared to talpids, tenrecids, and the marsupial Notoryctes. Correspondingly, the lack of such extensive coiling in the inner ear of other fossorial species may indicate a greater reliance on other senses to enable their fossorial lifestyle, such as tactile sensation from vibrissae and Eimer's organs. The reliance of chrysochlorids on sound is evident in the high degree of coiling and in the diversity of its mallear types, and may help explain the lack of any semiaquatic members of that group. The simplest mallear types among chrysochlorids are not present in the basal‐most members of that clade, but all extant chrysochlorids investigated to date exhibit extensive cochlear coiling. The chrysochlorid ear region thus exhibits mosaic evolution; our data suggest that extensive coiling evolved in chrysochlorids prior to and independently of diversification in middle ear ossicle size and shape. J. Morphol. 276:900–914, 2015. © 2015 Wiley Periodicals, Inc.     

Bone‐conduction hearing and seismic sensitivity of the late permian anomodont Kawingasaurus fossilis

An investigation of the internal cranial anatomy of the anomodont Kawingasaurus from the Upper Permian Usili Formation in Tanzania by means of neutron tomography revealed an unusual inner and middle ear anatomy such as extraordinarily inflated vestibules, lateroventrally orientated stapes with large footplates, and a small angle between the planes of the anterior and lateral semicircular canals. The vestibule has a volume, which is about 25 times larger than the human vestibule, although Kawingasaurus has only a skull length of approximately 40 mm. Vestibule inflation and enlarged stapes footplates are thought to be functionally correlated with bone‐conduction hearing; both morphologies have been observed in fossorial vertebrates using seismic signals for communication. The firmly fused triangular head with spatulate snout was probably used for digging and preadapted to seismic signal detection. The quadrate‐quadratojugal complex was able to transmit sound from the articular to the stapes by small vibrations of the quadrate process, which formed a ball and socket joint with the squamosal. Mechanical considerations suggest that the ventrolaterally orientated stapes of Kawingasaurus was mechanically better suited to transmit seismic sound from the ground to the fenestra vestibuli than a horizontal orientated stapes. The low sound pressure level transformer ratio of 2–3 in Kawingasaurus points to a seismic sensitivity of the middle ear and a vestigial or reduced sensitivity to airborne sound. Three hypothetical pathways of bone conduction in Kawingasaurus are discussed: 1) sound transmission via the spatulate snout and skull roof to the otic capsules, 2) relative movements resulting from the inertia of the mandible if sound is percepted with the skull, and 3) bone conduction from the substrate via mandible, jaw articulation, and stapes to the inner ear. J. Morphol. 276:121–143, 2015. © 2014 Wiley Periodicals, Inc.     

A Late Role for bmp2b in the Morphogenesis of Semicircular Canal Ducts in the Zebrafish Inner Ear

Background

The Bone Morphogenetic Protein (BMP) genes bmp2 and bmp4 are expressed in highly conserved patterns in the developing vertebrate inner ear. It has, however, proved difficult to elucidate the function of BMPs during ear development as mutations in these genes cause early embryonic lethality. Previous studies using conditional approaches in mouse and chicken have shown that Bmp4 has a role in semicircular canal and crista development, but there is currently no direct evidence for the role of Bmp2 in the developing inner ear.

Methodology/Principal Findings

We have used an RNA rescue strategy to test the role of bmp2b in the zebrafish inner ear directly. Injection of bmp2b or smad5 mRNA into homozygous mutant swirl (bmp2b^−/−) embryos rescues the early patterning defects in these mutants and the fish survive to adulthood. As injected RNA will only last, at most, for the first few days of embryogenesis, all later development occurs in the absence of bmp2b function. Although rescued swirl adult fish are viable, they have balance defects suggestive of vestibular dysfunction. Analysis of the inner ears of these fish reveals a total absence of semicircular canal ducts, structures involved in the detection of angular motion. All other regions of the ear, including the ampullae and cristae, are present and appear normal. Early stages of otic development in rescued swirl embryos are also normal.

Conclusions/Significance

Our findings demonstrate a critical late role for bmp2b in the morphogenesis of semicircular canals in the zebrafish inner ear. This is the first demonstration of a developmental role for any gene during post-embryonic stages of otic morphogenesis in the zebrafish. Despite differences in the early stages of semicircular canal formation between zebrafish and amniotes, the role of Bmp2 in semicircular canal duct outgrowth is likely to be conserved between different vertebrate species.     

The primate subarcuate fossa and its relationship to the semicircular canals part II: adult interspecific variation

Studies have reported an empirical link between the size of the semicircular canals and locomotor agility across adult primates. In this paper, we investigate the possibility that this relationship does not follow from the function of the semicircular canals to sense head rotations, but rather reflects spatial constraints imposed by the subarcuate fossa. The latter sits among the three canals and contains the petrosal lobule of the cerebellar paraflocculus, a structure involved in neural processing of locomotion-related eye movements. Hence, it is feasible that agility-related variations of lobule and fossa size affect the arc size of the surrounding semicircular canals. The present study tests such hypothetical correlations by evaluating canal size, fossa size, and agility among extant adult primates. Phylogenetically informed multivariate regression analyses show that, after controlling for body mass, the size of the subarcuate fossa has a significant positive effect on the overall size of the anterior canal and the width of the posterior canal. Multivariate regressions involving the height of the posterior canal and overall size of the lateral canal are not significant. Further bivariate analyses confirm that fossa size is unlikely to play a role in the previously reported link between agility and the size of the posterior and lateral canals. However, fossa size, especially its opening though the arc of the anterior canal, cannot be excluded as a factor that influences the size of the anterior canal more than agility. The findings show that the most reliable functional signals pertaining to locomotion in species that possess a patent subarcuate fossa are likely to come from the lateral canal and are least likely to come from the anterior canal.     

3D computational imaging of the petrosal of a new multituberculate mammal from the Late Cretaceous of China and its paleobiologic inferences

The derived middle and inner ears of mammals are the major features distinguishing them from non-mammalian vertebrates. Among them, multituberculate mammals represent an important transitional stage and a groundplan for further therian ear evolution. We present the reconstruction of petrosal features of a new multituberculate from the Late Cretaceous of Inner Mongolia (China) based on high resolution computed tomography and three-dimensional imaging analysis. Besides questioning some aspects of previous interpretations, this study reveals a combination of derived and primitive characters, such as a therian-like vascular and nervous pattern and internal acoustic meatus, and a monotreme-like inner ear, but with a derived semicircular canal planarity. The possible presence of a primary bony lamina for the basilar membrane could demonstrate that the first step in the elaboration of a coiled cochlea was already present in multituberculates. Auditory capabilities can be deduced for this animal, which was certainly terrestrial and possibly fossorial.     

The petrosal and inner ear of the Late Jurassic cladotherian mammal Dryolestes leiriensis and implications for ear evolution in therian mammals

Dryolestes leiriensis is a Late Jurassic fossil mammal of the dryolestoid superfamily in the cladotherian clade that includes the extant marsupials and placentals. We used high resolution micro‐computed tomography (µCT) scanning and digital reconstruction of the virtual endocast of the inner ear to show that its cochlear canal is coiled through 270°, and has a cribriform plate with the spiral cochlear nerve foramina between the internal acoustic meatus and the cochlear bony labyrinth. The cochlear canal has the primary bony lamina for the basilar membrane with a partially formed (or partially preserved) canal for the cochlear spiral ganglion. These structures, in their fully developed condition, form the modiolus (the bony spiral structure) of the fully coiled cochlea in extant marsupial and placental mammals. The CT data show that the secondary bony lamina is present, although less developed than in another dryolestoid Henkelotherium and in the prototribosphenidan Vincelestes. The presence of the primary bony lamina with spiral ganglion canal suggests a dense and finely distributed cochlear nerve innervation of the hair cells for improved resolution of sound frequencies. The primary, and very probably also the secondary, bony laminae are correlated with a more rigid support for the basilar membrane and a narrower width of this membrane, both of which are key soft‐tissue characteristics for more sensitive hearing for higher frequency sound. All these cochlear features originated prior to the full coiling of the therian mammal cochlea beyond one full turn, suggesting that the adaptation to hearing a wider range of sound frequencies, especially higher frequencies with refined resolution, has an ancient evolutionary origin no later than the Late Jurassic in therian evolution. The petrosal of Dryolestes has added several features that are not preserved in the petrosal of Henkelotherium. The petrosal characters of dryolestoid mammals are essentially the same as those of Vincelestes, helping to corroborate the synapomorphies of the cladotherian clade in neural, vascular, and other petrosal characteristics. The petrosal characteristics of Dryolestes and Henkelotherium together represent the ancestral morphotype of the cladotherian clade (Dryolestoidea + Vincelestes + extant Theria) from which the extant therian mammals evolved their ear region characteristics. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 166 , 433–463.