Ecogeographic patterns in fetal limb proportions.

OBJECTIVES: Humans generally comply with the ecological rule of Allen (1877), with populations from tropical environments exhibiting body proportions in which limb segments are long relative to trunk height compared to temperate groups. This study tests whether ecogeographic differences in intralimb proportions are identifiable among two modern fetal samples of differing ancestry. MATERIALS AND METHODS: Data are derived from radiographic measurements of long bone diaphyseal length and crown-heel length (CHL) of contemporary, spontaneously aborted fetuses of African Americans ("black") of assumed African (tropical) ancestry and European Americans ("white") of assumed European (temperate) ancestry (n = 184). Population individual limb elements, brachial, and crural indices are compared via analyses of covariance (ANCOVA). Potential patterns of divergent allometric growth are quantified through principal components analysis (PCA). RESULTS: African ancestral distal limb elements were consistently, albeit slightly, longer than those of European ancestry, relative to CHL. None of the ANCOVA interactions with ancestry are statistically significant for limb indices. The radius was the only single element that displayed a statistically significant ancestry effect (p = 0.0435) equating to a 1 mm difference. PCA highlights that upper limbs demonstrate negative allometry and lower limbs demonstrate positive allometry with sample-specific multivariate growth patterns being nearly identical. Differences in growth allometry late in gestation make little contribution to observed differences in adult limb proportions. DISCUSSION: No statistically significant ecogeographic patterns were appreciated among intralimb proportions between these groups during the fetal period. This study contributes to a greater appreciation of phenotypic plasticity, ecogeographic variation in ontogeny, and the evolution of modern human diversity.

The Influence of trisomy 21 on facial form and variability.

Triplication of chromosome 21 (trisomy 21) results in Down syndrome (DS), the most common live-born human aneuploidy. Individuals with DS have a unique facial appearance that can include form changes and altered variability. Using 3D photogrammatic images, 3D coordinate locations of 20 anatomical landmarks, and Euclidean Distance Matrix Analysis methods, we quantitatively test the hypothesis that children with DS (n = 55) exhibit facial form and variance differences relative to two different age-matched (4-12 years) control samples of euploid individuals: biological siblings of individuals with DS (n = 55) and euploid individuals without a sibling with DS (n = 55). Approximately 36% of measurements differ significantly between DS and DS-sibling samples, whereas 46% differ significantly between DS and unrelated control samples. Nearly 14% of measurements differ significantly in variance between DS and DS sibling samples, while 18% of measurements differ significantly in variance between DS and unrelated euploid control samples. Of those measures that showed a significant difference in variance, all were relatively increased in the sample of DS individuals. These results indicate that faces of children with DS are quantitatively more similar to their siblings than to unrelated euploid individuals and exhibit consistent, but slightly increased variation with most individuals falling within the range of normal variation established by euploid samples. These observations provide indirect evidence of the strength of the genetic underpinnings of the resemblance between relatives and the resistance of craniofacial development to genetic perturbations caused by trisomy 21, while underscoring the complexity of the genotype-phenotype map.

Craniofacial divergence by distinct prenatal growth patterns in Fgfr2 mutant mice.

BACKGROUND: Differences in cranial morphology arise due to changes in fundamental cell processes like migration, proliferation, differentiation and cell death driven by genetic programs. Signaling between fibroblast growth factors (FGFs) and their receptors (FGFRs) affect these processes during head development and mutations in FGFRs result in congenital diseases including FGFR-related craniosynostosis syndromes. Current research in model organisms focuses primarily on how these mutations change cell function local to sutures under the hypothesis that prematurely closing cranial sutures contribute to skull dysmorphogenesis. Though these studies have provided fundamentally important information contributing to the understanding of craniosynostosis conditions, knowledge of changes in cell function local to the sutures leave change in overall three-dimensional cranial morphology largely unexplained. Here we investigate growth of the skull in two inbred mouse models each carrying one of two gain-of-function mutations in FGFR2 on neighboring amino acids (S252W and P253R) that in humans cause Apert syndrome, one of the most severe FGFR-related craniosynostosis syndromes. We examine late embryonic skull development and suture patency in Fgfr2 Apert syndrome mice between embryonic day 17.5 and birth and quantify the effects of these mutations on 3D skull morphology, suture patency and growth. RESULTS: We show in mice what studies in humans can only infer: specific cranial growth deviations occur prenatally and worsen with time in organisms carrying these FGFR2 mutations. We demonstrate that: 1) distinct skull morphologies of each mutation group are established by E17.5; 2) cranial suture patency patterns differ between mice carrying these mutations and their unaffected littermates; 3) the prenatal skull grows differently in each mutation group; and 4) unique Fgfr2-related cranial morphologies are exacerbated by late embryonic growth patterns. CONCLUSIONS: Our analysis of mutation-driven changes in cranial growth provides a previously missing piece of knowledge necessary for explaining variation in emergent cranial morphologies and may ultimately be helpful in managing human cases carrying these same mutations. This information is critical to the understanding of craniofacial development, disease and evolution and may contribute to the evaluation of incipient therapeutic strategies.

Trisomy 21 and facial developmental instability.

The most common live-born human aneuploidy is trisomy 21, which causes Down syndrome (DS). Dosage imbalance of genes on chromosome 21 (Hsa21) affects complex gene-regulatory interactions and alters development to produce a wide range of phenotypes, including characteristic facial dysmorphology. Little is known about how trisomy 21 alters craniofacial morphogenesis to create this characteristic appearance. Proponents of the "amplified developmental instability" hypothesis argue that trisomy 21 causes a generalized genetic imbalance that disrupts evolutionarily conserved developmental pathways by decreasing developmental homeostasis and precision throughout development. Based on this model, we test the hypothesis that DS faces exhibit increased developmental instability relative to euploid individuals. Developmental instability was assessed by a statistical analysis of fluctuating asymmetry. We compared the magnitude and patterns of fluctuating asymmetry among siblings using three-dimensional coordinate locations of 20 anatomic landmarks collected from facial surface reconstructions in four age-matched samples ranging from 4 to 12 years: (1) DS individuals (n = 55); (2) biological siblings of DS individuals (n = 55); 3) and 4) two samples of typically developing individuals (n = 55 for each sample), who are euploid siblings and age-matched to the DS individuals and their euploid siblings (samples 1 and 2). Identification in the DS sample of facial prominences exhibiting increased fluctuating asymmetry during facial morphogenesis provides evidence for increased developmental instability in DS faces. We found the highest developmental instability in facial structures derived from the mandibular prominence and lowest in facial regions derived from the frontal prominence.

Technical note: a landmark-based approach to the study of the ear ossicles using ultra-high-resolution X-ray computed tomography data.

Previous study of the ear ossicles in Primates has demonstrated that they vary on both functional and phylogenetic bases. Such studies have generally employed two-dimensional linear measurements rather than three-dimensional data. The availability of Ultra- high-resolution X-ray computed tomography (UhrCT) has made it possible to accurately image the ossicles so that broadly accepted methodologies for acquiring and studying morphometric data can be applied. Using UhrCT data also allows for the ossicular chain to be studied in anatomical position, so that it is possible to consider the spatial and size relationships of all three bones. One issue impeding the morphometric study of the ear ossicles is a lack of broadly recognized landmarks. Distinguishing landmarks on the ossicles is difficult in part because there are only two areas of articulation in the ossicular chain, one of which (the malleus/incus articulation) has a complex three-dimensional form. A measurement error study is presented demonstrating that a suite of 16 landmarks can be precisely located on reconstructions of the ossicles from UhrCT data. Estimates of measurement error showed that most landmarks were highly replicable, with an average CV for associated interlandmark distances of less than 3%. The positions of these landmarks are chosen to reflect not only the overall shape of the bones in the chain and their relative positions, but also functional parameters. This study should provide a basis for further examination of the smallest bones in the body in three dimensions.

Phenotypic integration of neurocranium and brain.

Evolutionary history of Mammalia provides strong evidence that the morphology of skull and brain change jointly in evolution. Formation and development of brain and skull co-occur and are dependent upon a series of morphogenetic and patterning processes driven by genes and their regulatory programs. Our current concept of skull and brain as separate tissues results in distinct analyses of these tissues by most researchers. In this study, we use 3D computed tomography and magnetic resonance images of pediatric individuals diagnosed with premature closure of cranial sutures (craniosynostosis) to investigate phenotypic relationships between the brain and skull. It has been demonstrated previously that the skull and brain acquire characteristic dysmorphologies in isolated craniosynostosis, but relatively little is known of the developmental interactions that produce these anomalies. Our comparative analysis of phenotypic integration of brain and skull in premature closure of the sagittal and the right coronal sutures demonstrates that brain and skull are strongly integrated and that the significant differences in patterns of association do not occur local to the prematurely closed suture. We posit that the current focus on the suture as the basis for this condition may identify a proximate, but not the ultimate cause for these conditions. Given that premature suture closure reduces the number of cranial bones, and that a persistent loss of skull bones is demonstrated over the approximately 150 million years of synapsid evolution, craniosynostosis may serve as an informative model for evolution of the mammalian skull.

Technical note: out-of-plane angular correction based on a trigonometric function for use in two-dimensional kinematic studies.

In two-dimensional (2D) kinematic studies, limb positions in three-dimensional (3D) space observed in lateral view are projected onto a 2D film plane. Elbow and knee-joint angles that are less than 20 degrees out-of-plane of lateral-view cameras generally exhibit very little measurable difference from their 3D counterparts (Plagenhoef 1979 Environment, Behavior, and Morphology; New York: Gustav Fisher, p. 95-118). However, when limb segment angles are more than 20 degrees out-of-plane, as is often the case in locomotor studies of arboreal primates, elbow and knee angles can appear significantly more extended than they actually are. For this reason, a methodology is described that corrects 2D out-of-plane angular estimates using a series of trigonometric transformations.

Mandibular form in a rabbit model of familial nonsyndromic coronal suture synostosis.

Nonsyndromic coronal suture synostosis produces predictable and well-documented morphologies of the cranial vault with anteroposterior growth restrictions and mediolateral compensatory growth. The potential effects of nonsyndromic coronal suture synostosis on mandibular form are not as clear, however. This study was designed to evaluate whether coronal suture synostosis is associated with alterations in mandibular form by using a familial rabbit model of coronal suture synostosis. To assess this potential relation, the following hypothesis was tested: mandibular form in rabbits with coronal suture synostosis is significantly (P < 0.05) different from that seen in normal rabbits. The cleaned and dried mandibles of 33 adult New Zealand white rabbits were used (12 from normal rabbits, 13 from rabbits with delayed-onset synostosis, and 8 from rabbits with complete coronal suture synostosis). Seven anatomical landmarks on the mandible were located and digitized in three dimensions: anterior molar on the alveolus, posterior molar on the alveolus, coronoid process, anterior pole of the condyle, condylar process, angular process, and mandibular angle. To describe the mandibular condyle, the distance from the anterior pole to the posterior pole of the condyle was measured with digital sliding calipers, as was the distance between the medial and lateral poles. A shape ratio was then created using the dividend of these sums. Statistical analyses of mean form differences between mandibles were executed using Euclidean distance matrix analysis. Statistical analyses of the mandibular condyle linear and shape measurements were analyzed using one-way ANOVA in the three groups. Results showed that complete coronal suture synostosis is associated with significant (P < 0.05) differences in mandibular form compared with that of normal rabbits but that mandibular form in rabbits with delayed-onset synostosis does not differ from that of normal rabbits (P > 0.05). In particular, distances involving the coronoid process in rabbits with coronal suture synostosis were significantly different, paralleling previous work in human patients with coronal synostosis. There are no intrinsic condylar linear or shape differences between any of these groups, however. The form difference noted is most likely secondary to the synostosed coronal suture and may reflect alterations in the cranial base or masticatory musculature in this rabbit model.