Ontogenetic and functional modularity in the rodent mandible

The material properties of diets consumed by juvenile individuals are known to affect adult morphological outcomes. However, much of the current experimental knowledge regarding dietary effects on masticatory form is derived from studies in which individuals are fed a non-variable diet for the duration of their postweaning growth period. Thus, it remains unclear how intra-individual variation in diet, due to ontogenetic variation in feeding behaviors or seasonal resource fluctuations, affects the resulting adult morphology. Furthermore, the mandible is composed of multiple developmental and functional subunits, and the extent to which growth and plasticity among these modules is correlated may be misestimated by studies that examine non-variable masticatory function in adults only. To address these gaps in our current knowledge, this study raised Sprague Dawley rats (n = 42) in four dietary cohorts from weaning to skeletal maturity. Two cohorts were fed a stable (“annual”) diet of either solid or powdered pellets. The other two cohorts were fed a variable (“seasonal”) diet consisting of solid/powdered pellets for the first half of the study, followed by a shift to the opposite diet. Results of longitudinal morphometric analyses indicate that variation in the mandibular corpus is more prominent at immature ontogenetic stages, likely due to processes of dental eruption and the growth of tooth roots. Furthermore, adult morphology is influenced by both masticatory function and the ontogenetic timing of this function, e.g., the consumption of a mechanically resistant diet. The morphology of the coronoid process was found to separate cohorts on the basis of their early weanling diet, suggesting that the coronoid process/temporalis muscle module may have an early plasticity window related to high growth rates during this life stage. Conversely, the morphology of the angular process was found to be influenced by the consumption of a mechanically resistant diet at any point during the growth period, but with a tendency to reflect the most recent diet. The prolonged plasticity window of the angular process/pterygomasseteric muscle module may be related to delayed ossification and muscular maturation within this module. The research presented here highlights the importance of more naturalistic models of mammalian feeding, and underscores the need for a better understanding of the processes of both morphological and behavioral maturation that follow weaning

Adaptations to tree-gouging in the anterior masticatory apparatus of marmosets (callithrix) [abstract]

Although all genera of Callitrichinae engage in exudativory to some degree, marmosets (Callithrix, Cebuella) take advantage of exudates to the greatest extent. To facilitate exudate feeding, marmosets use their anterior teeth to gouge holes in bark and actively stimulate gum flow. As such, their anterior mandibular teethpossess specialized adaptations such as thickened labial enamel. Marmosets alsoshow masticatory features that facilitate increased gape, but do not appear to generate relatively large bite forces during gouging. However, even without increased bite force the anterior teeth of gougers likely experience different loading patternscompared to non-gouging platyrrhines. Specifically, one might expect that theanterior teeth and symphysis of marmosets are adapted to accommodate relatively high stresses linked to dissipating forces from yield-resistant and tough tree barks. This study uses histological data from thin- sectioned teeth, microCT data of jaws and teeth, and macroscale tests of simulated symphyseal loads to compare the micro- and macro-architecture of the anterior masticatory apparatus in Callithrix and Saguinus (as well as the outgroup Saimiri). Callithrix differs from the other genera in that its canine enamel possesses a much higher degree of decussation, and its anterior tooth roots are larger relative to alveolar bone volume. However, simulated jaw loading suggests a reduced ability to withstand external forces in the marmoset symphysis. The contrast between increased load-resistance ability in the anterior dentition versus relatively reduced symphyseal strength suggests both a potentially complex loading environment during gouging and a mosaic pattern of dentofacial adaptations to this derived biting behavior

Limitations of a morphological criterion of adaptive inference in the fossil record

Experimental analyses directly inform how an anatomical feature or complex functions during an organism's lifetime, which serves to increase the efficacy of comparative studies of living and fossil taxa. In the mammalian skull, food material properties and feeding behaviour have a pronounced influence on the development of the masticatory apparatus. Diet-related variation in loading magnitude and frequency induce a cascade of changes at the gross, tissue, cellular, protein and genetic levels, with such modelling and remodelling maintaining the integrity of oral structures vis-à-vis routine masticatory stresses. Ongoing integrative research using rabbit and rat models of long-term masticatory plasticity offers unique insight into the limitations of functional interpretations of fossilised remains. Given the general restriction of the palaeontological record to bony elements, we argue that failure to account for the disparity in the hierarchical network of responses of hard versus soft tissues may overestimate the magnitude of the adaptive divergence that is inferred from phenotypic differences. Second, we note that the developmental onset and duration of a loading stimulus associated with a given feeding behaviour can impart large effects on patterns of intraspecific variation that can mirror differences observed among taxa. Indeed, plasticity data are relevant to understanding evolutionary transformations because rabbits raised on different diets exhibit levels of morphological disparity comparable to those found between closely related primate species that vary in diet. Lastly, pronounced variation in joint form, and even joint function, can also characterise adult conspecifics that differ solely in age. In sum, our analyses emphasise the importance of a multi-site and hierarchical approach to understanding determinants of morphological variation, one which incorporates critical data on performance

Chewed Out: An Experimental Link between Food Material Properties and Repetitive Loading of the Masticatory Apparatus in Mammals

Using a model organism (rabbits) that resembles a number of mammalian herbivores in key aspects of its chewing behaviors, we examined how variation in dietary mechanical properties a ects food breakdown during mastication. Such data have implications for understanding phenotypic variation in the mammalian feeding apparatus, particularly with respect to linking jaw form to diet-induced repetitive loading. Results indicate that chewing frequency (chews/s) is independent of food properties, whereas chewing investment (chews/g) and chewing duration(s), which are proportional to repetitive loading of the jaws, are positively related to food sti ness and toughness. In comparisons of displacement-limited and stress-limited fragmentation indices, which respectively characterize the intraoral breakdown of tough and sti foods, increases in chewing investment and duration are linked solely to sti ness. This suggests that sti er foods engender higher peak loads and increased cyclical loading. Our ndings challenge conventional wisdom by demonstrating that toughness does not, by itself, underlie increases in cyclical loading and loading duration. Instead, tough foods may be associated with such jaw-loading patterns because they must be processed in greater volumes owing to their lower nutritive quality and for longer periods of time to increase oral exposure to salivary chemicals

Experimental Perspective on Fallback Foods and Dietary Adaptations in Early Hominins

The robust jaws and large, thick-enameled molars of the Plio–Pleistocene hominins Australopithecus and Paranthropus have long been interpreted as adaptations for hard-object feeding. Recent studies of dental microwear indicate that only Paranthropus robustus regularly ate hard items, suggesting that the dentognathic anatomy of other australopiths reflects rare, seasonal exploitation of hard fallback foods. Here, we show that hard-object feeding cannot explain the extreme morphology of Paranthropus boisei. Rather, analysis of long-term dietary plasticity in an animal model suggests year-round reliance on tough foods requiring prolonged postcanine processing in P. boisei. Increased consumption of such items may have marked the earlier transition from Ardipithecus to Australopithecus, with routine hard-object feeding in P. robustus representing a novel behaviour

Masticatory loading and soft-tissue plasticity in the mammalian circumorbital region

Abstract only availableMasticatory loading is the key to understanding the plasticity or epigenetic responses of many of the soft and hard tissues of the mammalian skull. Diet-induced variation in the magnitude and/or frequency of masticatory loads influences the organization, functional adaptation and postnatal development of craniofacial systems. While experimental work exists concerning masticatory stress as a determinant of maxillomandibular form, similar research concerning the mammalian circumorbital region of soft-tissue structures remains sparse. Indeed, controversy remains over the function of the circumorbital region, specifically whether the postorbital region in primates and other mammals is responsive to biomechanical loading. This hinders our understanding of evolutionary transformations during primate origins, where the earliest primates evolved a bony postorbital bar from an ancestor with a soft-tissue structure along the lateral orbital margin. To fill this gap, we examined the postorbital microanatomy of white rabbits (Oryctolagus cuniculus). Rabbits exhibit a masticatory complex and feeding behaviors like primates, yet retain a primitive circumorbital region similar to their common ancestor. To address the plasticity and function of soft tissues of the lateral orbital wall, three cohorts of 10 rabbits each were raised from weaning (1 month old) to adulthood (6 months old) on diets of different mechanical properties (under-use, control, over-use). Once sacrificed, tissues were collected from the left lateral orbital wall. Dissections revealed that, rather than the anticipated postorbital ligament, rabbits instead exhibit fibrocartilage. Samples were analyzed histologically and immunohistochemically for general anatomy, fibril architecture, collagen expression/organization, and protein abundance using Western Blot analysis. Preliminary data suggests that collagen fibers are aligned differently, with an overexpression of collagen in the over-use dietary cohort versus both the control and under-use groups. In sum, these experimental findings suggest that the postorbital fibrocartilage is mechanically responsive, which contrasts with the non-masticatory nature of bony elements in the circumorbital region.Life Sciences Undergraduate Research Opportunity Progra

Teaching an Old Jaw New Tricks: Diet-Induced Plasticity in a Model Organism, from Weaning to Adulthood

Many organisms exhibit a decrease in the ability to modify their phenotypes in response to shifts in environmental conditions as they mature. Such age-dependent plasticity has important implications in a variety of evolutionary and ecological contexts, particularly with respect to understanding adaptive responses to heterogeneous environments. In this study we used experimental diet manipulation to examine the life-history trajectory of plasticity in the feeding complex of a model organism, the white rabbit (Oryctolagus cuniculus). We demonstrate that, contrary to expectations derived from previous cross-sectional studies of skeletal plasticity, the jaws of weanlings and young adults exhibit similar increases in relative bone cross-sectional areas in response to the introduction of mechanically challenging foods into their diets. Furthermore, we present evidence that sensitivity to loading patterns persists well into adulthood in some regions of the masticatory apparatus in rabbits, indicating that there is an extended window of opportunity to respond to changes in dietary properties during an animal\u27s life span. We conclude that certain aspects of the facial skeleton of rabbits, and perhaps mammals in general, are sensitive to environmental stimuli long after skeletal maturity is achieved, highlighting the importance of plasticity as a source of adaptive variation at later life-history stages

The fracture toughness of small animal cortical bone measured using arc-shaped tension specimens: Effects of bisphosphonate and deproteinization treatments

Small animal models, and especially transgenic models, have become widespread in the study of bone mechanobiology and metabolic bone disease, but test methods for measuring fracture toughness on multiple replicates or at multiple locations within a single small animal bone are lacking. Therefore, the objective of this study was to develop a method to measure cortical bone fracture toughness in multiple specimens and locations along the diaphysis of small animal bones. Arc-shaped tension specimens were prepared from the mid-diaphysis of rabbit ulnae and loaded to failure to measure the radial fracture toughness in multiple replicates per bone. The test specimen dimensions, crack length, and maximum load met requirements for measuring the plane strain fracture toughness. Experimental groups included a control group, bisphosphonate treatment group, and an ex vivo deproteinization treatment following bisphosphonate treatment (5 rabbits/group and 15 specimens/group). The fracture toughness of ulnar cortical bone from rabbits treated with zoledronic acid for six months exhibited no difference compared with the control group. Partially deproteinized specimens exhibited significantly lower fracture toughness compared with both the control and bisphosphonate treatment groups. The deproteinization treatment increased tissue mineral density (TMD) and resulted in a negative linear correlation between the measured fracture toughness and TMD. Fracture toughness measurements were repeatable with a coefficient of variation of 12–16% within experimental groups. Retrospective power analysis of the control and deproteinization treatment groups indicated a minimum detectable difference of 0.1 MPa·m1/2. Therefore, the overall results of this study suggest that arc-shaped tension specimens offer an advantageous new method for measuring the fracture toughness in small animal bones

Non-sutural basicranium-derived cells undergo a unique mineralization pathway via a cartilage intermediate in vitro

The basicranium serves as a key interface in the mammalian skull, interacting with the calvarium, facial skeleton and vertebral column. Despite its critical function, little is known about basicranial bone formation, particularly on a cellular level. The goal of this study was therefore to cultivate a better understanding of basicranial development by isolating and characterizing the osteogenic potential of cells from the neonatal murine cranial base. Osteoblast-like basicranial cells were isolated, seeded in multicellular aggregates (designated micromasses), and cultured in osteogenic medium in the presence or absence of bone morphogenetic protein-6 (BMP6). A minimal osteogenic response was observed in control osteogenic medium, while BMP6 treatment induced a chondrogenic response followed by up-regulation of osteogenic markers and extensive mineralization. This response appears to be distinct from prior analyses of the calvarium and long bones, as basicranial cells did not mineralize under standard osteogenic conditions, but rather required BMP6 to stimulate mineralization, which occurred via an endochondral-like process. These findings suggest that this site may be unique compared to other cranial elements as well as the limb skeleton, and we propose that the distinct characteristics of these cells may be a function of the distinct properties of the basicranium: endochondral ossification, dual embryology, and complex loading environment