Morphological correspondence between brain and endocranial surfaces in mice exposed to undernutrition during development.

The morphological changes of the brain and the skull are highly integrated as a result of shared developmental pathways and different types of interactions between them. Shared developmental trajectories between these two structures might be influenced by genetic and environmental factors. Although the effect of environmental factors on neural and craniofacial traits has been extensively studied, less is known about the specific impact of stressful conditions on the coordinated variation between these structures. Here, we test the effect of early nutrient restriction on morphological correspondence between the brain and the endocast. For this purpose, mice exposed to protein or calorie-protein restriction during gestation and lactation were compared with a control group in which dams were fed standard food ad libitum. High-resolution images were obtained after weaning to describe brain and endocranial morphology. By magnetic resonance imaging (MRI), brain volumes were obtained and endocasts were segmented from skull reconstructions derived from micro-computed tomography (microCT). Brain and endocranial volumes were compared to assess the correspondence in size. Shape changes were analyzed using a set of landmarks and semilandmarks on 3D surfaces. Results indicated that brain volume is relatively less affected by undernutrition during development than endocast volume. Shape covariation between the brain and the endocast was found to be quite singular for protein-restricted animals. Procrustes distances were larger between the brain and the endocast of the same specimens than between brains or endocasts of different animals, which means that the greatest similarity is by type of structure and suggests that the use of the endocast as a direct proxy of the brain at this intraspecific scale could have some limitations. In the same line, patterns of brain shape asymmetry were not directly estimated from endocranial surfaces. In sum, our findings indicate that morphological variation and association between the brain and the endocast is modulated by environmental factors and support the idea that head morphogenesis results from complex processes that are sensitive to the pervasive influence of nutrient intake.

Association between shape changes and bone remodeling patterns in the middle face during ontogeny in South American populations.

The morphology of facial bones is modeled by processes of bone formation and resorption induced by interactions between tissues and compensatory responses. However, the role of remodeling patterns on the morphological changes within and among populations has been scarcely explored. Here, we assess the association between facial shape and the underlying bone cell activity throughout the ontogeny in two Amerindian populations that represent the extremes of craniofacial variation in South America. The sample comprises 71 individuals (36 adults and 35 subadults) representing hunter-gatherers from Patagonia and horticulturists from Northwest Argentina. We analyzed the shape and size of the zygomatic and the maxilla, and compared them with the patterns of bone formation and resorption. Bone formation and resorption were described by quantitative histological analysis of bone surfaces. Morphological changes were described by landmarks and semilandmarks digitized on 3D surfaces obtained from CT images. The results from multivariate statistics analysis show that the patterns of bone remodeling are associated with variation in the morphology of the middle face. We found a similar pattern of facial shape variation along the ontogenetic trajectory in the two samples, and a similar trend in the amount of formation and resorption activities across ages. The main differences between samples were found in the distribution of the areas of bone formation and resorption, possibly associated with mechanical bone response to masticatory loading. These findings provide clues about the processes and mechanisms of bone development involved in the facial morphological differentiation in human populations from southern South America.

Modeling the effect of brain growth on cranial bones using finite-element analysis and geometric morphometrics.

PURPOSE: Brain expansion during ontogeny has been identified as a key factor for explaining the growth pattern of neurocranial bones. However, the dynamics of this relation are only partially understood and a detailed characterization of integrated morphological changes of the brain and the neurocranium along ontogeny is still lacking. The aim of this study was to model the effect of brain growth on cranial bones by means of finite-element analysis (FEA) and geometric morphometric techniques. METHODS: First, we described the postnatal changes in brain size and shape by digitizing coordinates of 3D semilandmarks on cranial endocasts, as a proxy of brain, segmented from CT-scans of an ontogenetic sample. Then, two scenarios of brain growth were simulated: one in which brain volume increases with the same magnitude in all directions, and other that includes the information on the relative expansion of brain regions obtained from morphometric analysis. RESULTS: Results indicate that in the first model, in which a uniform pressure is applied, the largest displacements were localized in the sutures, especially in the anterior and posterior fontanels, as well as the metopic suture. When information of brain relative growth was introduced into the model, displacements were also concentrated in the lambda region although the values along both sides of the neurocranium (parietal and temporal bones) were larger than under the first scenario. CONCLUSION: In sum, we propose a realistic approach to the use of FEA based on morphometric data that offered different results to more simplified models.

Region-specific changes in Mus musculus brain size and cell composition under chronic nutrient restriction.

Nutrition is one of the most influential environmental factors affecting the development of different tissues and organs. It is suggested that under nutrient restriction the growth of the brain is spared as a result of the differential allocation of resources from other organs. However, it is not clear whether this sparing occurs brain-wide. Here, we analyzed morphological changes and cell composition in different regions of the offspring mouse brain after maternal exposure to nutrient restriction during pregnancy and lactation. Using high-resolution magnetic resonance imaging, we found that brain regions were differentially sensitive to maternal protein restriction and exhibited particular patterns of volume reduction. The cerebellum was reduced in absolute and relative volume, while cortex volume was relatively preserved. Alterations in cell composition (examined by the isotropic fractionator method) and organization of white matter (measured by diffusor tensor images) were also region specific. These changes were not related to the metabolic rate of the regions and were only partially explained by their specific growth trajectories. This study is a first step towards understanding the mechanisms of regional brain sparing at microstructural and macrostructural levels resulting from undernutrition.

Variation in facial bone growth remodeling in prehistoric populations from southern South America.

OBJECTIVES: To assess the intraspecific variation in bone remodeling patterns in modern humans, we studied two populations from southern South America that represent the extremes of morphological variation in this region. We particularly analyzed the ontogenetic changes in the patterns of bone growth remodeling and compared the patterns between samples. MATERIALS AND METHODS: We obtained high-resolution casts of the periosteal surface of the upper and middle face of subadults (n = 36) and adult (n = 36) individuals from a sample of hunter-gatherers from Patagonia and a sample of horticulturists from Northwest Argentina. The areas of bone formation and resorption were registered using an incident-light microscope. We then estimated the average bone remodeling map by sample and age, and performed principal component analysis and multivariate regressions to assess the extension and distribution of these areas across ontogeny and between samples. RESULTS: We found that the remodeling pattern of the glabella, supraorbital arch, frontal process of the maxilla, and a large part of the zygomatic bone is relatively constant in subadults and adults from both sample with a clear predominance of bone formation. In contrast, the middle face is characterized by the spatial alternation between formation and resorption areas, and greater variation with age and between samples. The main differences were found in areas related to chewing and muscle insertions. CONCLUSIONS: Our study provides the first evidence of interpopulation variation in bone growth remodeling and suggests that biomechanical factors can influence the observed patterns. It also underlines the need to account for ecological factors in within and between species comparisons.

Mitochondrial DNA Diversity and Evolutionary History of Native Human Populations of Argentinean Northwest Patagonia.

The genetic composition of Amerindian descendants from Patagonia has long been a focus of interest, although the information available is still scarce for many geographic areas. Here, we report the first analysis of the variation in the mitochondrial DNA (mtDNA) control region for an area of northwestern Patagonia, the North of Neuquén, with the aim of studying the processes and historical events that modeled the evolutionary history of these human groups. We analyzed 113 individuals from two localities of northern Neuquén, along with 6 from southern Neuquén and 223 previously published mtDNA sequences from neighboring areas in Argentina and Chile. We estimated the haplotypic variation and spatial structure of molecular variability. Amerindian subhaplogroups predominate in the two samples from northern Neuquén (n = 70), with D1g and C1b13 the most represented, although in different proportions. These samples exhibit Amerindian mtDNA haplotypes similar to the variants from neighboring areas. Most of haplotype variability was within group; variation among groups was relatively low and scarcely associated with geographical space. The most frequent subhaplogroups in northern Neuquén are characteristic of native populations from Patagonia and Chilean Araucanía, and probably originated in the region during the Late Pleistocene or Early Holocene. However, the spatial variation of mtDNA haplotypes departs from a latitudinal pattern and suggests differential levels of gene flow among areas during the Late Holocene, with moderate levels across the North of Neuquén as well as between this area and neighboring populations from Chile, the South of Neuquén, and Río Negro.

Integrating data on bone modeling and morphological ontogenetic changes of the maxilla in modern humans.

The aim of this work is to assess the association between the patterns of bone modeling and the changes in shape and size of the maxilla along ontogeny in modern humans. The sample analyzed includes an ontogenetic series of 30 individuals from an archeological site from Pampa Grande, northwest of Argentina. The areas of bone resorption and formation were described by histological analysis of bone surfaces and then quantified using spatial statistics. Morphological changes were analyzed by geometric morphometric methods using landmarks and semilandmarks digitized on 3D surfaces obtained from CT-scans. The regression of bone modeling maps on the centroid size shows no significant association between both variables neither in subadult nor adult individuals. On the contrary, the results of the partial least squares analysis shows a strong association between the shape changes in the maxilla with changes in the pattern of bone modeling in both groups of age, subadults and adults. Overall, this study contributes to the understanding of the mechanisms and processes that model maxillary morphology during growth.

Brain Structural Networks in Mouse Exposed to Chronic Maternal Undernutrition.

Brain structural connectivity is known to be altered in cases of intrauterine growth restriction and premature birth, although the specific effect of maternal nutritional restriction, a common burden in human populations, has not been assessed yet. Here we analyze the effects of maternal undernutrition during pregnancy and lactation by establishing three experimental groups of female mice divided according to their diet: control (Co), moderate calorie-protein restriction (MCP) and severe protein restriction (SP). Nutritionally restricted dams gained relatively less weight during pregnancy and the body weight of the offspring was also affected by maternal undernutrition, showing global growth restriction. We performed magnetic resonance imaging (MRI) of the offspring's brains after weaning and analyzed their connectivity patterns using complex graph theory. In general, changes observed in the MCP group were more subtle than in SP. Results indicated that brain structures were not homogeneously affected by early nutritional stress. In particular, the growth of central brain regions, such as the temporo-parietal cortex, and long integrative myelinated tracts were relatively preserved, while the frequency of short tracts was relatively reduced. We also found a differential effect on network parameters: network degree, clustering, characteristic path length and small-worldness remained mainly unchanged, while the rich-club index was lower in nutritionally restricted animals. Rich-club decrease reflects an impairment in the structure by which brain regions with large number of connections tend to be more densely linked among themselves. Overall, the findings presented here support the hypothesis that chronic nutritional stress produces long-term changes in brain structural connectivity.

A quantitative approach for analysing bone modelling patterns from craniofacial surfaces in hominins.

Bone size and shape arise throughout ontogeny as a result of the coordinated activity of osteoblasts and osteoclasts, responsible for bone deposition and resorption, and growth displacements. The modelling processes leave specific microstructural features on the bone surface, which can be used to infer the mechanisms shaping craniofacial traits in extinct and extant species. However, the analysis of bone surfaces from fossils and archaeological samples faces some difficulties related to the bone loss caused by taphonomic factors, and the lack of formal methods for estimating missing information and comparing the patterns of bone modelling among several specimens and samples. The present study provides a new approach for the quantitative analysis of bone formation and resorption patterns obtained from craniofacial surfaces. First, interpolation techniques were used to estimate missing data on high-resolution replicas of the left maxilla in a sample of sub-adult and adult modern humans and sub-adult fossil hominins. The performance of this approach was assessed by simulating variable amounts of missing data. Then, we applied measures of dispersion and central tendency to represent the variation and average pattern of bone modelling within samples. The spatial interpolation resulted in reliable estimations of the type of cell activity (deposition or resorption) in the missing areas, even when large extensions of the bone surface were lost. The quantification of the histological data allowed us to integrate the information of different specimens and depict the areas with higher and lower variation in the bone modelling pattern of the maxilla among specimens. Overall, the main advantages of the quantitative approach used here for generating bone modelling patterns are the high replicability and the possibility of incorporating variation among specimens into the comparisons among samples.

Morphological asymmetries of mouse brain assessed by geometric morphometric analysis of MRI data.

Mammalian brain has repeated structures at both sides of the median plane, although some asymmetries have been described even under normal conditions. Characterizing normal patterns of asymmetry in mouse brain is important to recognize features that depart from expected ranges in the most widely used mammalian model. Analyses on brain morphology based on magnetic resonance image (MRI) have largely focused on volumes while less is known about shape asymmetry. We introduce a flexible protocol based on geometric morphometrics to assess patterns of asymmetry in shape and size of mouse brain from microMRI scans. After systematic digitization of landmarks and semilandmarks, we combine multivariate methods for statistical analyses with visualization tools to display the results. No preliminary treatment of the images (e.g. space normalization) is needed to collect data on MRI slices and visual representations improve the interpretation of the results. Results indicated that the protocol is highly repeatable. Asymmetry was more evident for shape than for size. Particularly, fluctuating asymmetry accounted for more variation than directional asymmetry in all brain regions. Since this approach can detect subtle shape variation between sides, it is a promising methodology to explore morphological changes in the brain of model organisms and can be applied in future studies addressing the effect of genetic and environmental factors on brain morphology.

Technical note: Performance of semi and fully automated approaches for registration of 3D surface coordinates in geometric morphometric studies.

OBJECTIVES: One of the biggest challenges in the study of complex morphologies is to adequately describe shape variation. Here, we assess how the random sampling of surface points automatically obtained performs, when compared with observer-guided sampling procedures, and also evaluate the effect of sliding surface points by bending energy and minimum Procrustes distance. MATERIAL AND METHODS: Three datasets comprising structures with disparate levels of complexity and intrasample variation are as follows: mouse molars, mouse brains, and primate endocasts. Different configurations of 3D coordinates on curves and surfaces were digitized from MRI images and CT scans using semi and fully automated procedures. Shape variables were obtained by Generalized Procrustes Superpositions before and after sliding the pseudolandmarks. Multivariate analyses were used to summarize and compare shape variation. RESULTS: For the primate endocast, the semiautomated and automated strategies yield similar ordinations of specimens. Conversely, the semiautomated strategy better discriminates molar shapes between mouse groups. Shape differences among specimens are not adequately represented by the PCs calculated with surface pseudolandmarks. This is improved when the points are converted into semilandmarks by a sliding criterion. DISCUSSION: Surface semilandmarks automatically obtained from 3D models are promising although they should be used with some caution in complex structures. This approach can be taken as complementary of semiautomated procedures which perform better for assessing shape variation in localized traits previously selected while automated procedures are suitable in studies aimed at comparing overall variation in shape and when there is no previous information about highly variable anatomical regions.

Bone modeling patterns and morphometric craniofacial variation in individuals from two prehistoric human populations from Argentina.

Native human populations from South America display high levels of craniofacial variation encompassing gracile and robust skulls. Nevertheless, the processes of bone modeling by which morphological variation among populations were attained, remain poorly understood. Here we analyze the relationship between patterns of bone formation and resorption and morphometric variation in the upper face of adults belonging to farmers and hunter-gatherers from northwestern and south Argentina. Our analyses reveal a common pattern of bone modeling of the malar bone characterized by the presence of formation areas. Thus, the larger size and greater development of malar bone exhibited by hunter-gatherers would be linked to a greater magnitude of bone formation activity. Conversely, the glabella and the superciliary arch presented both formation and resorption areas with a variable distribution among individuals. In the extreme corresponding to more robust morphologies, the great development of the glabella is related to the presence of large formation fields, both in the upper region and toward the frontonasal suture. The less robust morphologies show resorption fields at the upper margin of the glabella, which would contribute to the weaker development of this region. The superciliary arch showed a complex relationship between its morphometric and histological variation; the individuals located at both extremes of the shape space presented large resorption areas located on its upper margin. Overall, our results show the existence of intraspecific variation in the patterns of bone modeling in the human upper face.

Effect of bite force and diet composition on craniofacial diversification of Southern South American human populations.

Ecological factors can be important to shape the patterns of morphological variation among human populations. Particularly, diet plays a fundamental role in craniofacial variation due to both the effect of the nutritional status-mostly dependent on the type and amount of nutrients consumed-on skeletal growth and the localized effects of masticatory forces. We examine these two dimensions of diet and evaluate their influence on morphological diversification of human populations from southern South America during the late Holocene. Cranial morphology was measured as 3D coordinates defining the face, base and vault. Size, form, and shape variables were obtained for 474 adult individuals coming from 12 samples. Diet composition was inferred from carious lesions and δ(13) C data, whereas bite forces were estimated using traits of main jaw muscles. The spatial structure of the morphological and ecological variables was measured using correlograms. The influence of diet composition and bite force on morphometric variation was estimated by a spatial regression model. Cranial variation and diet composition display a geographical structure, while no geographical pattern was observed in bite forces. Cranial variation in size and form is significantly associated with diet composition, suggesting a strong effect of systemic factors on cranial growth. Conversely, bite forces do not contribute significantly to the pattern of morphological variation among the samples analyzed. Overall, these results show that an association between diet composition and hardness cannot be assumed, and highlight the complex relationship between morphological diversification and diet in human populations.

Craniofacial variation, body size and ecological factors in aboriginal populations from central Patagonia (2000-200 years B.P.).

Previous studies have shown that ecological factors had a significant role in shaping the patterns of craniofacial variation among South American populations. Here, we evaluate whether temperature and diet contributed to facial diversification in small geographic areas. Facial size and shape of 9 osteological samples from central Patagonia (Argentina) were described using 2D landmarks and semilandmarks. Data on mean annual temperature, diet composition (δ(13)C and δ(15)N values) and femoral head maximum breadth, used as a proxy of body mass, were obtained for each sample. We then tested the association of body mass and the ecological variables with facial morphology using spatial regression techniques and a model selection approach. Akaike Information Criterion produced disparate results for both components of facial morphology. The best model for facial size included temperature and body mass proxy, and accounted for more than 80% of variation in size. Lower temperatures were related to larger facial sizes. Body mass was negatively associated with facial size and showed no relationship with the temperature. This suggests a relatively independent variation of cranial traits and body mass at the spatial scale studied here. Facial shape was not associated with the temperature or diet composition, contrasting with the patterns observed at larger spatial scales. Our results point out that the effect of climatic variables on cranial traits might be a source of morphological differentiation not only at large scales but also in small geographic areas, and that size and shape display a differential preservation of environmental signals.

Current status and perspectives of the development of dental research in biological anthropology of Argentina: introduction and conclusions of the symposium.

This paper describes and discusses the research in the field of dental anthropology in Argentina. It has been presented at the symposium entitled "The development of dental research in Argentine Biological Anthropology: current status and perspectives", coordinated by the authors at the IX National Meeting of Biological Anthropology of Argentina, Puerto Madryn, 20th-23rd October 2009. The aim of the symposium was to present new results and future prospects of this discipline in the country and to create a forum for discussion of current research within this field. Six contributions that focused on the study of teeth from different perspectives and analysed bioarchaeological samples from different areas of Argentina (Central Highlands, Pampa and Patagonia) were presented. After the presentations, a discussion about the state of the art of dental research in the country was generated, in which the need for the generation of methodological consensus on the criteria for the evaluation of the variables considered was stated, so that research conducted in different areas can be compared. In short, the contributions of this symposium provide insights into the diversity of dental anthropology in contemporary Argentina and the potential of these types of studies to gain important information about biological and cultural aspects of the native populations in the country.

Ontogenetic allometry and cranial shape diversification among human populations from South America.

Modifications of ontogenetic allometries play an important role in patterning the shape differentiation among populations. This study evaluates the influence of size variation on craniofacial shape disparity among human populations from South America and assesses whether the morphological disparity observed at the interpopulation level resulted from a variable extension of the same ontogenetic allometry, or whether it arose as a result of divergences in the pattern of size-related shape changes. The size and shape of 282 adult and subadult crania were described by geometric morphometric-based techniques. Multivariate regressions were used to evaluate the influence of size on shape differentiation between and within populations, and phylogenetic comparative methods were used to take into account the shared evolutionary history among populations. The phylogenetic generalized least-squares models showed that size accounts for a significant amount of shape variation among populations for the vault and face but not for the base, suggesting that the three modules did not exhibit a uniform response to changes in overall growth. The common slope test indicated that patterns of evolutionary and ontogenetic allometry for the vault and face were similar and characterized by a heightening of the face and a lengthening of the vault with increasing size. The conservation of the same pattern of shape changes with size suggests that differences in the extent of growth contributed to the interpopulation cranial shape variation and that certain directions of morphological change were favored by the trait covariation along ontogeny.

Ecological and evolutionary factors in dental morphological diversification among modern human populations from southern South America.

The knowledge of processes involved in morphological variation requires the integrated analysis of evolutionary and ecological factors. Here, we investigate the factors responsible for dental variation among human populations from southern South America. The aim of this work is to test the correspondence of dental size and shape variation with geographical, molecular (i.e. mtDNA) and ecological (i.e. climate, diet and food preparation) variables employing comparative phylogenetic methods, which have not previously been extensively applied at a within-species level. The results of the Procrustes analysis show a significant association of shape variables with molecular distance and geography, whereas dental size is not associated with molecular or geographical distances among groups. Phylogenetic generalized least-squares analysis, which takes into account the evolutionary autocorrelation among populations, shows a significant relationship between dental size variation and diet, while temperature and pottery do not correspond with dental size or shape. Specifically, groups with diets rich in carbohydrates, as well as the maritime hunter-gatherers, have the smallest teeth. In summary, our results support ecological factors as the dominant factor on dental size diversification in this region, while evolutionary relationships account for variation in dental shape.

Spatial patterns and evolutionary processes in southern South America: a study of dental morphometric variation.

The purpose of this article is to examine the patterns of evolutionary relationships between human populations from the later Late Holocene (1,500-100 years BP) of southern South America on the basis of dental morphometric data. We tested the hypotheses that the variation observed in this region would be explained by the existence of populations with different phylogenetic origin or differential action of gene flow and genetic drift. In this study, we analyzed permanent teeth from 17 samples of male and female adult individuals from throughout southern South America. We measured mesiodistal and buccolingual diameters at the base of the crown, along the cement-enamel junction. The results of multiple regression analysis and a mantel correlogram indicate the existence of spatial structure in dental shape variation, as the D(2) Mahalanobis distance between samples increases with increasing geographical distance between them. In addition, the correlation test results show a trend toward reduction of the internal variation of samples with increasing latitude. The detected pattern of dental variation agrees with the one expected as an outcome of founder serial effects related to an expansion of range during the initial occupation of southern South America.

Ontogeny of robusticity of craniofacial traits in modern humans: a study of South American populations.

To date, differences in craniofacial robusticity among modern and fossil humans have been primarily addressed by analyzing adult individuals; thus, the developmental basis of such differentiation remains poorly understood. This article aims to analyze the ontogenetic development of craniofacial robusticity in human populations from South America. Geometric morphometric methods were used to describe cranial traits in lateral view by using landmarks and semilandmarks. We compare the patterns of variation among populations obtained with subadults and adults to determine whether population-specific differences are evident at early postnatal ontogeny, compare ontogenetic allometric trajectories to ascertain whether changes in the ontogeny of shape contribute to the differentiation of adult morphologies, and estimate the amount of size change that occurs during growth along each population-specific trajectory. The results obtained indicate that the pattern of interpopulation variation in shape and size is already established at the age of 5 years, meaning that processes acting early during ontogeny contribute to the adult variation. The ontogenetic allometric trajectories are not parallel among all samples, suggesting the divergence in the size-related shape changes. Finally, the extension of ontogenetic trajectories also seems to contribute to shape variation observed among adults.