Fossil bears break free from inhibitory cascade constraints at least twice ( Ursus minimus and Ursus deningeri ) caused by dietary adaptations

The inhibitory cascade (IC) model is a widely used evolutionary developmental explanation of among-species differences in relative molar tooth size. The IC model posits that, as molars develop from front to back, the relative strength of activating and inhibiting influences establishes a “ratcheting” mechanism leading to predictable relative molar sizes. Such a constraint on molar covariation would lead to strong variational biases on the evolutionary paths that the molar row can traverse through phenotypic space. These constraints manifest themselves in characteristic patterns of variation among species that loosely match observed macroevolutionary patterns. In this paper, we write out the predictions of the IC model for within-species covariation in molar size in a framework that unifies evolutionary developmental biological and quantitative genetic perspectives on the evolution of complex traits. We then evaluate these predictions about aspects of molar covariation in eight anthropoid primate species. We find that the IC model tends to over-predict aspects of within-species covariation by substantial margins. Only macaques exhibit covariation in and among individual teeth consistent with the IC model, but they do not show signs of the strong evolutionary constraint predicted by the model. Gorillas meet none of the predictions. While we cannot rule out an IC-like process as a contributor, causes of molar size covariation other than those described in the IC model must be major contributors to covariation in molar teeth within populations.

BackgroundDevelopmental processes that underpin morphological variation have become a focus of interest when attempting to interpret macroevolutionary patterns. Recently, the Dental Inhibitory Cascade (dic) model has been suggested to explain much of the variation in mammalian molar size proportions. We tested the macroevolutionary implications of this model using anthropoid primate species (n=100), focusing on overall morphological patterns, as well as predictions made about molar size variability, direct developmental control, and diet.ResultsOf the species sampled, 56 % had centroids that fell within regions of molar proportion morphospace consistent with the dic model. We also found that the third molar had greater variation in size than either the first or second molars, as expected by the model. Some dic model predictions were not supported, however, such as the expected proportion of M2/M1 when the third molar is absent. Furthermore, we found that some variability in third molar size could not be explained by the influence of the inhibitory cascade. Overall, we found considerable clade-specific differences in relative molar sizes among anthropoid primates, with hominoids and cercopithecins strongly divergent from dic model predictions, and platyrrhines, colobines, and papionins more consistent with the inhibitory cascade. Finally, we investigated reasons why some clades deviated from dic model expectations. Adaptations for frugivory (e.g., bunodont cusp relief) appeared to be one driver of relatively larger second molars and have evolved independently in multiple lineages of anthropoids.ConclusionsThe dic model explains some of the variation in anthropoid primate molar proportions. However, there are interesting deviations away from this broad mammalian pattern, particularly in hominoids and cercopithecins, which suggest the model is only one of multiple mechanisms determining morphological variability in mammalian teeth.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-016-0673-5) contains supplementary material, which is available to authorized users.

SynopsisMammalian molar crowns form a module in which measurements of size for individual teeth within a tooth row covary with one another. Molar crown size covariation is proposed to fit the inhibitory cascade model (ICM) or its variant the molar module component (MMC) model, but the inability of the former model to fit across biological scales is a concern in the few cases where it has been tested in Primates. The ICM has thus far failed to explain patterns of intraspecific variation, an intermediate biological scale, even though it explains patterns at both smaller organ-level and larger between-species biological scales. Studies of this topic in a much broader range of taxa are needed, but the properties of a sample appropriate for testing the ICM at the intraspecific level are unclear. Here, we assess intraspecific variation in relative molar sizes of the cotton mouse, Peromyscus gossypinus, to further test the ICM and to develop recommendations for appropriate sampling protocols in future intraspecific studies of molar size variation across Mammalia. To develop these recommendations, we model the sensitivity of estimates of molar ratios to sample size and simulate the use of composite molar rows when complete ones are unavailable. Similar to past studies on primates, our results show that intraspecific variance structure of molar ratios within the rodent P. gossypinus does not meet predictions of the ICM or MMC. When we extend these analyses to include the MMC, one model does not fit observed patterns of variation better than the other. Standing variation in molar size ratios is relatively constant across mammalian samples containing all three molars. In future studies, analyzing average ratio values will require relatively small minimum sample sizes of two or more complete molar rows. Even composite-based estimates from four or more specimens per tooth position can accurately estimate mean molar ratios. Analyzing variance structure will require relatively large sample sizes of at least 40–50 complete specimens, and composite molar rows cannot accurately reconstruct variance structure of ratios in a sample. Based on these results, we propose guidelines for intraspecific studies of molar size covariation. In particular, we note that the suitability of composite specimens for averaging mean molar ratios is promising for the inclusion of isolated molars and incomplete molar rows from the fossil record in future studies of the evolution of molar modules, as long as variance structure is not a key component of such studies.

The implications of the inhibitory cascade (IC) model in dental diversification have been primarily studied at an interspecific or higher level. In contrast, the study of organisms with recent evolutionary divergence or at an interpopulational scale is still very limited. Here, we assess the effect of changes in molar size and the ratio of local activators to inhibitors on molar proportions based on a compilation of data of crown diameters of the first, second, and third lower and upper molars of extinct and extant hominids and modern human populations. The analysis of allometric changes between the size of each tooth and the size of the molar row shows a negative allometry in first molars (M1), isometric changes in second molars (M2), and a positive allometry in third molars (M3) in both hominin phylogeny and modern human populations. On the other hand, the proportions of lower and upper molars of several hominid species fall outside the morphospace defined by the IC model, while most of the modern human populations fall within the morphospace defined by the model as M1>M2>M3. We conclude that there is a phylogenetic structuring for molar size, particularly in the maxilla, with a trend toward mesial-to-distal reduction in the molar row area accompanied by allometric changes. Our findings also show the limitations of the IC model for explaining molar proportions in primates, particularly the variation in the relative size at the interspecific scale in the hominid lineage.

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.

Unexpected variation of human molar size patterns

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.

BackgroundMuch of the current research in the growing field of evolutionary development concerns relating developmental pathways to large-scale patterns of morphological evolution, with developmental constraints on variation, and hence diversity, a field of particular interest. Tooth morphology offers an excellent model system for such ‘evo-devo’ studies, because teeth are well preserved in the fossil record, and are commonly used in phylogenetic analyses and as ecological proxies. Moreover, tooth development is relatively well studied, and has provided several testable hypotheses of developmental influences on macroevolutionary patterns. The recently-described Inhibitory Cascade (IC) Model provides just such a hypothesis for mammalian lower molar evolution. Derived from experimental data, the IC Model suggests that a balance between mesenchymal activators and molar-derived inhibitors determines the size of the immediately posterior molar, predicting firstly that molars either decrease in size along the tooth row, or increase in size, or are all of equal size, and secondly that the second lower molar should occupy one third of lower molar area. Here, we tested the IC Model in a large selection of taxa from diverse extant and fossil mammalian groups, ranging from the Middle Jurassic (~176 to 161 Ma) to the Recent.ResultsResults show that most taxa (~65%) fell within the predicted areas of the Inhibitory Cascade Model. However, members of several extinct groups fell into the regions where m2 was largest, or rarely, smallest, including the majority of the polyphyletic “condylarths”. Most Mesozoic mammals fell near the centre of the space with equality of size in all three molars. The distribution of taxa was significantly clustered by diet and by phylogenetic group.ConclusionsOverall, the IC Model was supported as a plesiomorphic developmental system for Mammalia, suggesting that mammal tooth size has been subjected to this developmental constraint at least since the divergence of australosphenidans and boreosphenidans approximately 180 Ma. Although exceptions exist, including many ‘condylarths’, these are most likely to be secondarily derived states, rather than alternative ancestral developmental models for Mammalia.

The relative sizes of body segments are a major determinant of the shape and functionality of an animal. Developmental biases affecting this trait can therefore have major evolutionary implications. In vertebrates, a molecular activator/inhibitor mechanism, known as the inhibitory cascade (IC), produces a simple and predictable pattern of linear relative size along successive segments. The IC model is considered the default mode of vertebrate segment development and has produced long-term biases in the evolution of serially homologous structures such as teeth, vertebrae, limbs, and digits. Here we investigate whether the IC model or an IC-like model also has controls on segment size development in an ancient and hyperdiverse group of extinct arthropods, the trilobites. We examined segment size patterning in 128 trilobite species, and during ontogenetic growth in three trilobite species. Linear relative segment size patterning is prominent throughout the trunk of trilobites in the adult form, and there is strict regulation of this patterning in newly developing segments in the pygidium. Extending the analysis to select stem and modern arthropods suggests that the IC is a common default mode of segment development capable of producing long-term biases in morphological evolution across arthropods as it does in vertebrates.

The Middle Pleistocene Sima de los Huesos (SH) site has yielded more than 7.500 human fossil remains belonging to a minimum of 29 individuals. Most of these individuals preserve either the complete mandibular molar series or at least the first (M1 ) and second (M2 ) molars. The inhibitory cascade mathematical model was proposed by Kavanagh et al. (Nature, 449, 427-433 [2007]) after their experimental studies on the dental development of murine rodent species. The activator-inhibitor mechanism of this model has shown its ability for predicting evolutionary size patterns of mammalian teeth, including hominins. The main aim of this study is to test whether the size molar patterns observed in the SH hominins fit the inhibitory cascade model. With this purpose, we have measured the crown area of all SH molars in photographs, using a planimeter and following techniques used and well contrasted in previous works. Following one of the premises of the inhibitory cascade model, we expect that the central tooth (M2 in our case) of a triplet would have the average size of the two outer teeth. The absolute difference between the observed and the expected values for the M2 s ranges from 0.23 to 8.46mm2 in the SH sample. In terms of percentage, the difference ranges between 0.25% and 10.34%, although in most cases, it is below 5%. The plot of the estimated M3 /M1 and M2 /M1 size ratios obtained in the SH hominins occupies a small area of the theoretical developmental morphospace obtained for rodent species. In addition, the majority of the values are placed near the theoretical line which defines the relationship predicted by the inhibitory cascade model in these mammals. The values of the slope and intercept of the reduced major regression obtained for the SH individuals do not differ significantly from those obtained for rodent species, thus confirming that the size of the molars of the SH hominins fits the inhibitory cascade model. We discuss these results in terms of dental development. Despite the promising results in the SH sample, we draw the attention to the fact that most Early Pleistocene Homo specimens exhibit a pattern (M1 <M2 >M3 ), which is outside the expected theoretical morphospace predicted by the inhibitory cascade model. The shift from the M1<M2<M3 size relationship observed in early hominins (including H.habilis) to the M1>M2>M3 size relationship, which is predominant in modern humans, includes sequences that depart from predictions of the inhibitory cascade model. Additional studies are required to understand these deviations.

Macroscelidid afrotherians and canid carnivorans possess four premolar loci, the first of which is not replaced. Previous work suggests that the first premolar in macroscelidids is a retained deciduous tooth, but in Canis it is a successional tooth with no milk precursor. We tested this contrasting interpretation of first premolar homology with data from ontogenetic anatomy and with area predictions from the inhibitory cascade (IC) model. Our results based on anatomy support previous interpretations that the functional first premolar is a retained deciduous tooth (dp1) with no successor in macroscelidids, and a successional tooth (p1) with no precursor in Canis. Hyracoids are among the few placental mammals that show replacement at the first premolar locus and show less deviation than other taxa of actual from predicted areas across the deciduous and molar toothrow. However, predicted vs. actual tooth areas can depart substantially from one another. At least without a better means of representing tooth size, the inhibitory cascade does not help to distinguish the deciduous from successional first premolar. This observation does not rule out the possibility that factors such as a size-shift within the toothrow (e.g., carnivoran carnassials) help to explain deviations from the inhibitory cascade model.

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.

Molar size and diet in the Strepsirrhini: Implications for size-adjustment in studies of primate dental adaptation

The developmental processes that contribute to variation of morphological traits are the subject of considerable interest when attempting to understand phenotypic evolution. It is well demonstrated that most characteristics of tooth pattern can be modified by tinkering conserved signal pathways involved in dental development. This effect can be evaluated by comparing developmental models with naturally occurring variation within explicit phylogenetic contexts. Here, we assess whether evolutionary changes in lower molar (M) ratios among platyrrhines were channelled by alterations in the balance of activators and inhibitors as predicted by the inhibitory cascade (IC) model (Kavanagh et al. in Nature 449:427–432, 2007). Ordinary linear regression adjusted to M2/M1 versus M3/M1 ratios of 38 species of platyrrhines indicated that the slope and intercept were significantly different from the IC model. Conversely, when the phylogeny was incorporated into the regression analyses (PGLS), variation in molar ratios did not differ from the developmental model. PGLS also showed that changes in molar proportions are not an allometric effect associated with body size. Discrepancies between phylogenetically corrected and non-corrected analyses are mainly due to the departure of Callitrichines from the predicted values. This subfamily displays agenesis of M3 with higher than expected M2/M1 ratios, indicating that M3 fails to develop even when the inhibition by M1 on the subsequent molars is not increased. Our results show that evolution in molar ratios is concordant with slight changes in the proportion of activators and inhibitors that regulate molar development; however, other processes are required to account for variation in the number of teeth.

The Inhibitory Cascade Model (ICM) predicts in mice that a larger permanent first molar (M1) results in a smaller second molar (M2) and an even smaller third molar (M3). Counter to the ICM, our lab found in contemporary humans that instead of M1 size, only M2 size predicts M3 size. Our lab also identified in humans 13 different molar size ratio patterns instead of one classic ICM pattern (M1&gt;M2&gt;M3). Further, other work in nonhuman primates reported a Premolar Molar Module (PMM) where sizes of the fourth premolar (P4) and all three molars are highly correlated. However, premolar:molar size correlation in contemporary humans is yet to be explored. This exploration can help define how the development of these two different classes of teeth influence each other’s sizes. In humans, M1 initiates near birth, followed ~24 months later by P3 and P4 (almost simultaneously with each other), then ~12 months later by M2, and ~6 years later by M3. Our first aim was to define variation in premolar size ratio pattern. We hypothesized that all three mathematically possible premolar size ratio patterns (P3=P4, P3&gt;P4, P3&lt;P4) will be observed in our sample. Our second aim was to test whether premolar crown size and size ratio predict molar absolute sizes and molar size ratio patterns (e.g., M1&gt;M2&gt;M3). We hypothesized that absolute sizes as well as size ratios of earlier initiating teeth will predict the absolute sizes and size ratios of later initiating teeth. Maximum mesiodistal lengths of P3, P4, M1, M2, and M3 crowns were measured in millimeters to represent absolute tooth sizes using retrospective Cone Beam Computed Tomography scans of 91 dental patients (54 female, 37 male) aged 13‐23 years. A frequency analysis revealed all three mathematically possible premolar size ratio patterns in our sample (P3&gt;P4: 49.83%, P3&lt;P4: 34.13%, and P3=P4: 16.04%, n=293 oral quadrants). Also, 57% (n=49) of the subjects with data collected from all four quadrants showed all three premolar size ratio patterns within the same mouth. A linear mixed model analysis (95% confidence level, alpha = 0.05) showed that P3 and P4 predicted each other’s crown size, as well as that of M2, but not M3. M1 size predicts P4 size only. In sum, we found that premolar size ratio patterns are highly variable even within the same oral cavity. We also found strong correlations among P3, P4 and M2 sizes, and between M1 and P4 sizes. Our results found variability in premolar size ratio patterns that is similar to the variability that we previously found in molar size ratio patterns. Our study in contemporary humans does not support the ICM prediction of a single classic tooth size ratio pattern. However, our study outcomes supported the PMM findings with the exception that P4 was not correlated with M3.