Geographic Variation of Absolute and Relative Lower Molar Sizes in Two Closely Related Species of Japanese Field Mice (Apodemus speciosus and Apodemus argenteus: Muridae, Rodentia).

Geographic variation of the sizes of lower molar (M1 size) and relative lower molar sizes (size proportions among M1, M2, and M3) were examined in two species of closely related Japanese field mice (Apodemus speciosus and Apodemus argenteus). To determine the cause of the geographic variations observed, phylogeographic structure, interspecific competition, climate, and location (mainland or island) were compared. With regard to the phylogeographic structure, the sizes of the molar and the relative molar sizes in A. speciosus did not differ between two major clades (mainland vs. Hokkaido and peripheral islands), whereas the phylogeographic structure was not examined in A. argenteus, as no clear phylogeographic structure was evident. The sizes of M1 and relative molar size (M3/M1 score) in A. speciosus differed significantly between the mainland and islands; however, there was no significant difference between islands within and outside the distribution of A. argenteus. Interspecific competition between the two species may thus not be considerable. Climatic factors (temperature) and relative molar sizes (M2/M1 and M3/M1 scores) were significantly correlated in the mainland populations of A. speciosus, indicating that geographic variations in relative molar sizes may be affected by climate. In addition, M3/M1 scores varied more in the islands than on the mainland, suggesting effects of genetic drift. However, M1 size increases in the island populations of the two species are not attributed to the climate, but are explained by the so-called Island Rule. Geographic variation in A. speciosus is thus likely attributable to various effects.

Geographic Variation of Absolute and Relative Lower Molar Sizes in the Japanese Macaque (Macaca fuscata: Primates, Mammalia).

We examined geographic variations of absolute and relative lower molar sizes (size proportions among M1, M2, and M3) in the Japanese macaque (Macaca fuscata) using skull specimens obtained from 13 locations. We compared the geographic patterns and climatic factors. The size of M1 significantly and negatively correlated to the annual and coldest month mean temperatures and precipitation for both males and females. The M2/M1 and M3/M1 scores significantly and positively correlated to the annual and coldest month mean temperatures. The geographic pattern in the size of M1 was consistent with Bergmann's rule; however, the sizes of M2 and M3 did not correlate with temperature, and were not consistent with the rule. The geographic pattern in relative molar sizes (M2/M1 and M3/M1 scores) indicated that populations living in colder climates possess a larger M1 in relation to M2 and M3. Therefore, the correlations of M1, M2/M1, and M3/M1 scores to temperature involve an increase in the size of M1 in a colder climate. In macaques, the functions of the different molars (M1, M2, and M3) do not differ (they all exhibit grinding function, unlike differentiation between carnassial and other molars in Carnivora), whereas the timing of molar eruption does. In other words, at young ages (1.5-3.5 years), M1 erupts and is in occlusion, whereas M2 and M3 do not erupt and are not used for mastication. Therefore, the geographic pattern in the relative molar sizes may be attributed to increasing survivability in harsh winter climates by increasing occlusal surface in younger animals.

Effects of Usag-1 and Bmp7 deficiencies on murine tooth morphogenesis.

BACKGROUND: Wnt5a and Mrfzb1 genes are involved in the regulation of tooth size, and their expression levels are similar to that of Bmp7 during morphogenesis, including during the cap and early bell stages of tooth formation. We previously reported that Usag-1-deficient mice form supernumerary maxillary incisors. Thus, we hypothesized that BMP7 and USAG-1 signaling molecules may play important roles in tooth morphogenesis. In this study, we established double genetically modified mice to examine the in vivo inter-relationships between Bmp7 and Usag-1. RESULTS: We measured the volume and cross-sectional areas of the mandibular incisors using micro-computed tomography (micro-CT) in adult Bmp7- and Usag-1-LacZ knock-in mice and their F2 generation upon interbreeding. The mandibular incisors of adult Bmp7+/- mice were significantly larger than those of wild-type (WT) mice. The mandibular incisors of adult Usag-1-/- mice were the largest of all genotypes examined. In the F2 generation, the effects of these genes were additive; Bmp7+/- was most strongly associated with the increase in tooth size using generalized linear models, and the total area of mandibular supernumerary incisors of Usag-1-/-Bmp7+/- mice was significantly larger than that of Usag-1-/-Bmp7 +/+ mice. At embryonic day 15 (E15), BrdU assays demonstrated that the labeling index of Bmp7+/- embryos was significantly higher than that of WT embryos in the cervical loop. Additionally, the labeling index of Usag-1-/- embryos was significantly the highest of all genotypes examined in dental papilla. CONCLUSIONS: Bmp7 heterozygous mice exhibited significantly increased tooth sizes, suggesting that tooth size was controlled by specific gene expression. Our findings may be useful in applications of regenerative medicine and dentistry.

Shape variation in the skull and lower carnassial in a wild population of raccoon dog (Nyctereutes procyonoides).

Individual variations in skull and lower carnassial morphology within a wild population of raccoon dog were examined using geometric morphometric techniques. We compared individual morphological variations by using relative warp analysis, and then tested morphological integration between the skull and carnassial by using partial least square (PLS) analysis. The most marked variation in skull shape was the dorsoventral flexion; i.e., deformation from klinorhynchy to airorhynchy. Two remarkable variations were observed, including tilting between the trigonid (or carnassial blade) and the talonid in the lower carnassial, and the relative sizes of the trigonid and the talonid. This observed variation in skull shape was similar to previous reports of variations among dog breeds that correlate with a polymorphism of the Runx2 gene. This polymorphism has also been reported to correlate with snout length, which is strongly related to carnivorous or omnivorous dietary adaptations, across the entire order Carnivora. Our results in the lower carnassial were also similar to previously reported patterns observed for carnivorous or omnivorous dietary adaptations among Carnivora. However, in our PLS analysis between skull and carnassial shapes, we only found a significant correlation in a lower dimension, suggesting a lower degree of integration. These results indicate that shape variations, which could be sources of natural selection in the skull and carnassial, were present in a wild population, suggesting high evolvability of these variations in the raccoon dog and the order Carnivora in general.

Ontogenetic scaling of the humerus in sea turtles and its implications for locomotion.

In the present study, we analyzed the ontogenetic scaling of humeri in the green turtle (Chelonia mydas) and loggerhead turtle (Caretta caretta). Green turtles have relatively thicker humeri than loggerhead turtles, indicating that the humerus of the green turtle can resist greater loads. Our results are consistent with isometry, or slightly negative allometry, of diameter in relation to length of the humerus in both species. Geometric similarity or isometry of the humerus in relation to body mass is supported by estimates of the cross-sectional properties of green turtles. Sea turtles are adapted for aquatic life, but also perform terrestrial locomotion. Thus, during terrestrial locomotion, which requires support against gravity, the observed scaling relationships indicate that there may be greater stress and fracture risk on the humeri of larger green turtles than on the humeri of smaller turtles. In aquatic habitats, in which limbs are mainly used for propulsion, the stress and fracture risk for green turtle humeri are estimated to increase with greater speed. This scaling pattern may be related to the possibility that smaller turtles swim at a relatively faster speed per body length.

Revisiting the evolutionary trend toward the mammalian lower jaw in non-mammalian synapsids in a phylogenetic context.

The mammalian lower jaw comprises a single bone, the dentary, which is a unique feature among vertebrates. The lower jaws of extinct non-mammalian synapsids were composed of the dentary and several postdentary bones. Synapsid fossils exhibit variation in the dentary size relative to the overall lower jaw. An evolutionary trend toward dentary enlargement and postdentary reduction in non-mammalian synapsids has long been documented but has not been established using modern phylogenetic comparative methods. In this study, we examine the evolutionary pattern of dentary size relative to the lower jaw through phylogenetic analyses of measurements in a broad range of non-mammalian synapsid taxa. Our analyses revealed an evolutionary trend toward dentary area enlargement relative to the overall lower jaw in the lateral view across all non-mammalian synapsids. This trend is likely due to vertical expansion of the dentary given that the same trend is not evident when looking at anterior to posterior measurements of the dentary relative to the lower jaw as a whole in lateral view. Ancestral character reconstructions revealed that the evolution of the measurements was not unidirectional in non-mammalian synapsids. Our results provide no evidence of an evolutionary trend toward the dentary enlargement at the expense of postdentary bones across non-mammalian synapsids. This implies that the evolutionary origin of the mammalian lower jaw is not adequately explained by the evolutionary trend of dentary enlargement throughout non-mammalian synapsids. Instead, selection that occurred during the transition from non-mammalian cynodonts to early mammals may have produced the mammalian lower jaw.

The anteriorization of tooth position underlies the atavism of tooth morphology: Insights into the morphogenesis of mammalian molars.

The evolution and development of complex molars as a key innovation in mammals have long been of interest yet remain poorly understood. With reference to century-old theories and modern findings, we focused on the teeth of pinnipeds (Carnivora) and cetaceans (Cetartiodactyla), which are morphologically simple compared with those of other mammals, and thus can be considered a reversal toward the ancestral state of nonmammalian synapsids. By reconstructing the evolutionary history of tooth complexity for the phylogenies of Carnivora and Cetartiodactyla, we established that a secondary evolution of simple teeth from more complex molars has occurred independently multiple times. Our phylogenetic comparative analyses showed that a simplification in tooth morphology was correlated with a more anterior dentition position relative to the component bones of the upper jaw in both Carnivora and Cetartiodactyla. These results suggest that the anterior shift of tooth position relative to the morphogenetic fields present in the jaw contributed to the evolutionary simplification in molar morphology. Our findings provide insights into the developmental basis of complex mammalian dentition.

Sexual dimorphism in external morphology of the American bullfrog Rana (Aquarana) catesbeiana and the possibility of sex determination based on tympanic membrane/eye size ratio.

The American bullfrog Rana (Aquarana) catesbeiana has been reported to show significant sexual dimorphism based on the size ratio between the tympanic membrane and the eye. In males the tympanic membrane is much larger than the eye, but not in females. The ratio has been used as a convenient criterion to discriminate sexes (sexing) in the American bullfrog, though its reliability is unknown. In this study, we examined 86 adult American bullfrogs to clarify whether the tympanic membrane long diameter/eye long diameter (Dtm/De) ratio is a reliable index to discriminate sexes in this species. In addition, we examined the growth of this sexually dimorphic trait. Results indicated that there is a significant difference but there is a small overlap in this ratio Dtm/De between sexes. The allometric comparisons showed the sexual dimorphism of the Dtm/De ratio was increased during growth and the dimorphism is attributable to the difference in the growth rate of the tympanic membrane (Dtm). Therefore, sex determination of American bullfrogs cannot be wholly reliably achieved by the Dtm/De ratio alone; other external morphological features are required in addition.

Dental integration and modularity in pinnipeds.

Morphological integration and modularity are important for understanding phenotypic evolution because they constrain variation subjected to selection and enable independent evolution of functional and developmental units. We report dental integration and modularity in representative otariid (Eumetopias jubatus, Callorhinus ursinus) and phocid (Phoca largha, Histriophoca fasciata) species of Pinnipedia. This is the first study of integration and modularity in a secondarily simplified dentition with simple occlusion. Integration was stronger in both otariid species than in either phocid species and related positively to dental occlusion and negatively to both modularity and tooth-size variability across all the species. The canines and third upper incisor were most strongly integrated, comprising a module that likely serves as occlusal guides for the postcanines. There was no or weak modularity among tooth classes. The reported integration is stronger than or similar to that in mammals with complex dentition and refined occlusion. We hypothesise that this strong integration is driven by dental occlusion, and that it is enabled by reduction of modularity that constrains overall integration in complex dentitions. We propose that modularity was reduced in pinnipeds during the transition to aquatic life in association with the origin of pierce-feeding and loss of mastication caused by underwater feeding.

Loss of Stemness, EMT, and Supernumerary Tooth Formation in Cebpb-/-Runx2+/- Murine Incisors.

Adult Cebpb KO mice incisors present amelogenin-positive epithelium pearls, enamel and dentin allopathic hyperplasia, fewer Sox2-positive cells in labial cervical loop epitheliums, and reduced Sox2 expression in enamel epithelial stem cells. Thus, Cebpb acts upstream of Sox2 to regulate stemness. In this study, Cebpb KO mice demonstrated cementum-like hard tissue in dental pulp, loss of polarity by ameloblasts, enamel matrix in ameloblastic layer, and increased expression of epithelial-mesenchymal transition (EMT) markers in a Cebpb knockdown mouse enamel epithelial stem cell line. Runx2 knockdown in the cell line presented a similar expression pattern. Therefore, the EMT enabled disengaged odontogenic epithelial stem cells to develop supernumerary teeth. Cebpb and Runx2 knockdown in the cell line revealed higher Biglycan and Decorin expression, and Decorin-positive staining in the periapical region, indicating their involvement in supernumerary tooth formation. Cebpb and Runx2 acted synergistically and played an important role in the formation of supernumerary teeth in adult incisors.

Geographic variation of craniodental morphology of the Eurasian otter (Lutra lutra) in East Asia.

Craniodental morphology of the Eurasian otter (Lutra lutra) in the Korean Peninsula, Japanese islands and Kinmen Island (Taiwan) was studied using geometric morphometrics to identify the skull variations between the populations. Forty adult skulls were examined (29 specimens from the Korean Peninsula, six from Shikoku, Honshu and Hokkaido of Japan, and five from Kinmen Island). Images of the dorsal and ventral views of the skull and the right lateral view of the mandible were analyzed. Specimens from the Korean Peninsula were larger than those from the Japanese islands and Kinmen Island. However, no correlation was observed between the shape variations in the three populations and the centroid size of the skull. The Mann-Whitney U-test showed that relative warps (RWs) RW1, RW2 and RW4 of the dorsal view and RW2 of the ventral view of the skull differed significantly between the populations. Some craniodental differences between the populations were seen in the dorsal and ventral views of the skull, mostly at the snout and parietal regions. The MANOVA test revealed significant differences between the specimens from the Japanese islands and Korean Peninsula and between the specimens from the Korean Peninsula and Kinmen Island. RWs plots showed an overlap of all three populations. In conclusion, the comparisons of the three examined populations revealed significant differences in their craniodental morphology.

The origin of the lower fourth molar in canids, inferred by individual variation.

BACKGROUND: An increase in tooth number is an exception during mammalian evolution. The acquisition of the lower fourth molar in the bat-eared fox (Otocyon megalotis, Canidae, Carnivora, Mammalia) is one example; however, its developmental origin is not clear. In some canids (Canidae), individual variation exist as supernumerary molar M4. This study focuses on the acquisition of the lower fourth molar in canids and proposes that the inhibitory cascade model can explain its origin. METHODS: Occlusal view projected area of lower molars was determined from 740 mandibles obtained from Canis latrans, Nyctereutes procyonoides, and Urocyon cinereoargenteus museum specimens. For each molar, relative sizes of molars (M2/M1 and M3/M1 scores) affected by inhibition/activation dynamics during development, were compared between individuals with and without supernumerary molar (M4). RESULTS: Possession of a supernumerary molar was associated with significantly larger M2/M1 score in Canis latrans, M3/M1 score in Nyctereutes procyonoides, and M2/M1 and M3/M1 scores in Urocyon cinereoargenteus compared to individuals of these species that lacked supernumerary molars. DISCUSSION: We propose that, in canids, the supernumerary fourth molar is attributable to reduced inhibition and greater activation during molar development. In the bat-eared fox, altered inhibition and activation dynamics of dental development during omnivorous-insectivorous adaptation may be a contributing factor in the origin of the lower fourth molar.

Sexual dimorphism of the Eurasian otter (Lutra lutra) in South Korea: Craniodental geometric morphometry.

Sexual dimorphism of the craniodental morphology of the Eurasian otter in South Korea was studied with geometric morphometrics. 29 adult skulls (15 males and 14 females) were used. Images of the dorsal and ventral view of the cranium and right lateral view of the mandible were taken and then digitized, and measurements were taken on the right side. Results showed that size difference between males and females was significant. Correlations between the size and shape variations have not been observed in this study. The bivariate plots with centroid size showed size dimorphism between males and females with some overlapping. Most relative warp (RW) scores were not significantly different between males and females. We observed only RW2 for dorsal and ventral view of the skull, and only RW1 for mandible was significantly different between the sexes. Shape dimorphisms were revealed at the postorbital constriction, temporal-mandibular joint, coronoid process, mandibular condyle and angular process of the skull. Based on our study, sexual dimorphism exists in Eurasian otter from the South Korean population in terms of both the size and shape. Furthermore, the degree of size dimorphism is greater than shape dimorphism.

Comparative cranial morphology in living and extinct platypuses: Feeding behavior, electroreception, and loss of teeth.

The modern platypus, Ornithorhynchus anatinus, has an eye structure similar to aquatic mammals; however, platypuses also have a "sixth sense" associated with the bill electro- and mechanoreception that they use without opening their eyes underwater. We hypothesize that Ornithorhynchus and the Miocene taxon Obdurodon have different sensory capacities, which may have resulted from differences in foraging behavior. To estimate differences in foraging, sensory systems, and anatomical divergence between these monotremes, we compared their skull morphologies. Results indicate that the bill of Obdurodon is more dorsally deflected than that of Ornithorhynchus, suggesting a pelagic foraging behavior in Obdurodon compared to the bottom-feeding behavior in Ornithorhynchus. The infraorbital foramen of Obdurodon, through which the maxillary nerve passes sensory data from the bill to the brain, is relatively less developed than that of Ornithorhynchus. Whereas bill-focused sensory perception was likely shared among Mesozoic monotremes, the highly developed electrosensory system of Ornithorhynchus may represent an adaptation to foraging in cloudy water. Computed tomography imagery indicates that the enlarged infraorbital canal of Ornithorhynchus restricts the space available for maxillary tooth roots. Hence, loss of functional teeth in Ornithorhynchus may possibly have resulted from a shift in foraging behavior and coordinate elaboration of the electroreceptive sensory system. Well-developed electroreceptivity in monotremes is known at least as far back as the early Cretaceous; however, there are differences in the extent of elaboration of the feature among members of the ornithorhynchid lineage.

Tooth Size Variation in Pinniped Dentitions.

It is contentious whether size variation among mammalian teeth is heterogeneous or homogeneous, whether the coefficient of variation is reliable, and whether the standard deviation of log-transformed data and the residual of standard deviation on mean variable size are useful replacements for the coefficient of variation. Most studies of tooth size variation have been on mammals with complex-crowned teeth, with relatively little attention paid to taxa with simple-crowned teeth, such as Pinnipedia. To fill this gap in knowledge and to resolve the existing controversies, we explored the variation of linear size variables (length and width) for all teeth from complete permanent dentitions of four pinniped species, two phocids (Histriophoca fasciata, Phoca largha) and two otariids (Callorhinus ursinus, Eumetopias jubatus). Size variation among these teeth was mostly heterogeneous both along the toothrow and among species. The incisors, canines, and mesial and distal postcanines were often relatively highly variable. The levels of overall dental size variation ranged from relatively low as in land carnivorans (Phoca largha and both otariids) to high (Histriophoca fasciata). Sexual size dimorphism varied among teeth and among species, with teeth being, on average, larger in males than in females. This dimorphism was more pronounced, and the canines were larger and more dimorphic relative to other teeth in the otariids than in the phocids. The coefficient of variation quantified variation reliably in most cases. The standard deviation of log-transformed data was redundant with the coefficient of variation. The residual of standard deviation on mean variable size was inaccurate when size variation was considerably heterogeneous among the compared variables, and was incomparable between species and between sexes. The existing hypotheses invoking developmental fields, occlusal complexity, and the relative timing of tooth formation and sexually dimorphic hormonal activity do not adequately explain the differential size variation along the pinniped toothrow.

Unique inhibitory cascade pattern of molars in canids contributing to their potential to evolutionary plasticity of diet.

Developmental origins that guide the evolution of dental morphology and dental formulae are fundamental subjects in mammalian evolution. In a previous study, a developmental model termed the inhibitory cascade model was established. This model could explain variations in relative molar sizes and loss of the lower third molars, which sometimes reflect diet, in murine rodents and other mammals. Here, I investigated the pattern of relative molar sizes (inhibitory cascade pattern) in canids, a taxon exhibiting a wide range of dietary habits. I found that interspecific variation in canid molars suggests a unique inhibitory cascade pattern that differs from that in murine rodents and other previously reported mammals, and that this variation reflects dietary habits. This unique variability in molars was also observed in individual variation in canid species. According to these observations, canid species have greater variability in the relative sizes of first molars (carnassials), which are functionally important for dietary adaptation in the Carnivora. In conclusion, an inhibitory cascade that differs from that in murine rodents and other mammals may have contributed to diverse dietary patterns and to their parallel evolution in canids.