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Living primates show a complex trend in reduction of nasal cavity spaces and structures due to moderate to severe constraint on interorbital breadth. Here we describe the ontogeny of the posterior end of the primate cartilaginous nasal capsule, the thimble shaped posterior nasal cupula (PNC), which surrounds the hind end of the olfactory region. We used a histologically sectioned sample of strepsirrhine primates and two non-primates (Tupaia belangeri, Rousettus leschenaulti), and histochemical and immunohistochemical methods to study the PNC in a perinatal sample. At birth, most strepsirrhines possess only fragments of PNC, and these lack a perichondrium. Fetal specimens of several species reveal a more complete PNC, but the cartilage exhibits uneven or weak reactivity to type II collagen antibodies. Moreover, there is relatively less matrix than in the septal cartilage, resulting in clustering of chondrocytes, some of which are in direct contact with adjacent connective tissues. In one primate (Varecia spp.) and both non-primates, the PNC has a perichondrium at birth. In older, infant Varecia and Rousettus, the perichondrium of the PNC is absent, and PNC fragmentation at its posterior pole has occurred in the former. Loss of the perichondrium for the PNC appears to precede resorption of the posterior end of the nasal capsule. These results suggest that the consolidation of the basicranial and facial skeletons happens ontogenetically earlier in primates than other mammals. We hypothesize that early loss of cartilage at the sphenoethmoidal articulation limits chondral mechanisms for nasal complexity, such as interstitial expansion or endochondral ossification.

Cranial synchondroses are cartilaginous joints between basicranial bones or between basicranial bones and septal cartilage, and have been implicated as having a potential active role in determining craniofacial form. However, few studies have examined them histologically. Using histological and immunohistochemical methods, we examined all basicranial joints in serial sagittal sections of newborn heads from nine genera of primates (five anthropoids, four strepsirrhines). Each synchondrosis was examined for characteristics of active growth centers, including a zonal distribution of proliferating and hypertrophic chondrocytes, as well as corresponding changes in matrix characteristics (i.e., density and organization of Type II collagen). Results reveal three midline and three bilateral synchondroses possess attributes of active growth centers in all species (sphenooccipital, intrasphenoidal, presphenoseptal). One midline synchondrosis (ethmoseptal) and one bilateral synchondrosis (alibasisphenoidal synchondrosis [ABS]) are active growth centers in some but not all newborn primates. ABS is oriented more anteriorly in monkeys compared to lemurs and bushbabies. The sphenoethmoidal synchondrosis (SES) varies at birth: in monkeys, it is a suture-like joint (i.e., fibrous tissue between the two bones); however, in strepsirrhines, the jugum sphenoidale is ossified while the mesethmoid remains cartilaginous. No species possesses an SES that has the organization of a growth plate. Overall, our findings demonstrate that only four midline synchondroses have the potential to actively affect basicranial angularity and facial orientation during the perinatal timeframe, while the SES of anthropoids essentially transitions toward a "suture-like" function, permitting passive growth postnatally. Loss of cartilaginous continuity at SES and reorientation of ABS distinguish monkeys from strepsirrhines.

Ectotympanic bone morphology is commonly used as a large-scale phylogenetic indicator across primates. Less well-understood is the intraspecies variation in this characteristic and the dynamic ways in which it affects and is affected by overall basicranial shape. This study attempts to clarify relationships between the external ear canal and basicranial shape among humans in a sample of archaeological human crania. The ectotympanic length and orientation were hypothesized to correlate with the shape of the cranial base and particularly with shape variables associated with relative brachycephaly. Basicranial shape in 80 computed tomography (CT) scans of adult humans were quantified using landmark coordinate data, with particular emphasis on the cranial base and auditory structures. Scaled ectotympanic lengths were taken from interlandmark distances and then compared to shape variation of the whole basicranium as summarized by procrustes shape variables and principal components analysis (PCA). The length of the ectotympanic bone was correlated with total cranial base variation. Long ectotympanic bones were found to be associated with brachycephalic individuals and less flexed basicrania. Additionally, long ectotympanic bones were found to be more horizontally oriented, as opposed to inferiorly sloped. We suggest that as brachycephaly increases the distance between the otic capsule and the pinna, the ectotympanic bone lengthens in response.

Nasal turbinals, delicate and complex bones of the nasal cavity that support respiratory or olfactory mucosa (OM), are now easily studied using high resolution micro-computed tomography (μ-CT). Standard μ-CT currently lacks the capacity to identify OM or other mucosa types without additional radio-opaque staining techniques. However, even unstained mucosa is more radio-opaque than air, and thus mucosal thickness can be discerned. Here, we assess mucosal thickness of the nasal fossa using the cranium of a cadaveric adult dog that was μ-CT scanned with an isotropic resolution of 30 μm, and subsequently histologically sectioned and stained. After co-alignment of μ-CT slice planes to that of histology, mucosal thickness was estimated at four locations. Results based on either μ-CT or histology indicate olfactory mucosa is thicker on average compared with non-olfactory mucosa (non-OM). In addition, olfactory mucosa has a lesser degree of variability than the non-OM. Variability in the latter appears to relate mostly to the varying degree of vascularity of the lamina propria. Because of this, in structures with both specialized vascular respiratory mucosa and OM, such as the first ethmoturbinal (ET I), the range of thickness of OM and non-OM may overlap. Future work should assess the utility of diffusible iodine-based contrast enhanced CT techniques, which can differentiate epithelium from the lamina propria, to enhance our ability to differentiate mucosa types on more rostral ethmoturbinals. This is especially critical for structures such as ET I, which have mixed functional roles in many mammals.

The range of cranial morphology seen in domestic dogs (Canis lupus familiaris) is a direct result of thousands of years of selective breeding. This article is the first to investigate how selection for reduced faces in brachycephalic dogs impacted the neuroanatomy of the canine brain through the analysis of endocasts. Previous research has demonstrated global effects on the shape of the bony cranium as the result of these breeding practices; however, these studies have largely focused on the bony structures of the skull and failed to consider the influence of facial reduction on the soft tissues of the brain. We generated endocasts from an existing set of clinically-obtained CT scans representing a variety of dogs with various cranial morphologies. These dogs represented four breeds as well as a comparative sample of dogs of unknown breed. We recorded three-dimensional coordinate data for 31 landmarks representing various gyri, sulci, and other neuroanatomical landmarks that allowed us to analyze differences in shape of the endocasts. Through geometric morphometric analyses, we determined that the endocast shape variance in this sample is correlated with cephalic index, and thus the selection for facial reduction has caused a perceivable effect on canine neuroanatomy. Additionally, we found the majority of the shape variance in the sample to be associated with olfactory anatomy; however, the rest of the morphology also correlates with cephalic index. The results of this article indicate that modern breeding practices and the selection for dogs with short faces have significantly influenced canine neuroanatomy.

Previous descriptive work on deciduous dentition of primates has focused disproportionately on great apes and humans. To address this bias in the literature, we studied 131 subadult nonhominoid specimens (including 110 newborns) describing deciduous tooth morphology and assessing maximum hydroxyapatite density (MHD). All specimens were CT scanned at 70 kVp and reconstructed at 20.5–39 μm voxels. Grayscale intensity from scans was converted to hydroxyapatite (HA) density (mg HA/cm(3)) using a linear conversion of grayscale values to calibration standards of known HA density (R(2) = .99). Using Amira software, mineralized dental tissues were captured by segmenting the tooth cusps first and then capturing the remainder of the teeth at descending thresholds of gray levels. We assessed the relationship of MHD of selected teeth to cranial length using Pearson correlation coefficients. In monkeys, anterior teeth are more mineralized than postcanine teeth. In tarsiers and most lemurs and lorises, postcanine teeth are the most highly mineralized. This suggests that monkeys have a more prolonged process of dental mineralization that begins with incisors and canines, while mineralization of postcanine teeth is delayed. This may in part be a result of relatively late weaning in most anthropoid primates. Results also reveal that in lemurs and lorises, MHD of the mandibular first permanent molar (M(1)) negatively correlates with cranial length. In contrast, the MHD of M(1) positively correlates with cranial length in monkeys. This supports the hypothesis that natural selection acts independently on dental growth as opposed to mineralization and indicates clear phylogenetic differences among primates.

Diffusible iodine-based contrast-enhanced computed tomography (diceCT) has emerged as a viable tool for discriminating soft tissues in serial CT slices, which can then be used for three-dimensional analysis. This technique has some potential to supplant histology as a tool for identification of body tissues. Here, we studied the head of an adult fruit bat (Cynopterus sphinx) and a late fetal vampire bat (Desmodus rotundus) using diceCT and µCT. Subsequently, we decalcified, serially sectioned and stained the same heads. The two CT volumes were rotated so that the sectional plane of the slice series closely matched that of histological sections, yielding the ideal opportunity to relate CT observations to corresponding histology. Olfactory epithelium is typically thicker, on average, than respiratory epithelium in both bats. Thus, one investigator (SK), blind to the histological sections, examined the diceCT slice series for both bats and annotated changes in thickness of epithelium on the first ethmoturbinal (ET I), the roof of the nasal fossa, and the nasal septum. A second trial was conducted with an added criterion: radioopacity of the lamina propria as an indicator of Bowman’s glands. Then, a second investigator (TS) annotated images of matching histological sections based on microscopic observation of epithelial type, and transferred these annotations to matching CT slices. Measurements of slices annotated according to changes in epithelial thickness alone closely track measurements of slices based on histologically-informed annotations; matching histological sections confirm blind annotations were effective based on epithelial thickness alone, except for a patch of unusually thick non-OE, mistaken for OE in one of the specimens. When characteristics of the lamina propria were added in the second trial, the blind annotations excluded the thick non-OE. Moreover, in the fetal bat the use of evidence for Bowman’s glands improved detection of olfactory mucosa, perhaps because the epithelium itself was thin enough at its margins to escape detection. We conclude that diceCT can by itself be highly effective in identifying distribution of OE, especially where observations are confirmed by histology from at least one specimen of the species. Our findings also establish that iodine staining, followed by stain removal, does not interfere with subsequent histological staining of the same specimen.

The analysis of textiles as trace evidence is an important area of focus in the field of forensic science, because enhanced understanding of the decomposition of textiles may point to more accurate methods for estimating the post mortem interval (PMI) of remains found in association with these materials. This research is especially crucial in areas with unique climates, like the state of Florida. This study examines the generation of microclimates from the differential decomposition of various textile types. This study hypothesized that the decomposition of textiles will generate microclimates with soil properties that differ from those of the surrounding environment, and that different types of textiles will create different microclimates as they decompose. Samples of cotton, UV-proofed cotton, polyester, cotton-polyester blended fabric, ripstop, and wool were buried at four sites on a property in North Central Florida for thirteen weeks, with measures of soil temperature, pH, and moisture level, and weather data collected weekly. Following burial, decomposition of each textile type was scored. Data collected were analyzed in R statistical software. Analysis indicated that the level of degradation differed by textile type but not by site. Textile presence, type of textile, and subsequent decomposition significantly impacted soil pH and moisture at all sites, but did not have a significant effect on soil temperature. The results of this study demonstrate that the decomposition of textiles can create diverse and unique microclimates in the soil environment. When found in association with human remains, presence and type of textile should be considered when estimating decomposition rates and the postmortem interval.

Our knowledge of nasal cavity anatomy has grown considerably with the advent of micro-computed tomography (CT). More recently, a technique called diffusible iodine-based contrast-enhanced CT (diceCT) has rendered it possible to study nasal soft tissues. Using diceCT and histology, we aim to (a) explore the utility of these techniques for inferring the presence of venous sinuses that typify respiratory mucosa and (b) inquire whether distribution of vascular mucosa may relate to specialization for derived functions of the nasal cavity (i.e., nasal-emission of echolocation sounds) in bats. Matching histology and diceCT data indicate that diceCT can detect venous sinuses as either darkened, "empty" spaces, or radio-opaque islands when blood cells are present. Thus, we show that diceCT provides reliable information on vascular distribution in the mucosa of the nasal airways. Among the bats studied, a nonecholocating pteropodid (Cynopterus sphinx) and an oral-emitter of echolocation sounds (Eptesicus fuscus) possess venous sinus networks that drain into the sphenopalatine vein rostral to the nasopharynx. In contrast, nasopharyngeal passageways of nasal-emitting hipposiderids are notably packed with venous sinuses. The mucosae of the nasopharyngeal passageways are far less vascular in nasal-emitting phyllostomids, in which vascular mucosae are more widely distributed in the nasal cavity, and in some nectar-feeding species, a particularly large venous sinus is adjacent to the vomeronasal organ. Therefore, we do not find a common pattern of venous sinus distribution associated with nasal emission of sounds in phyllostomids and hipposiderids. Instead, vascular mucosa is more likely critical for air-conditioning and sometimes vomeronasal function in all bats.

The sphenoid bone articulates with multiple basicranial, facial, and calvarial bones, and in humans its synchondroses are known to contribute to elongation of the skull base and possibly to cranial base angulation. Its early development (embryological, early fetal) has frequently been studied in a comparative context. However, the perinatal events in morphogenesis of the sphenoid have been explored in very few primates. Using a cross-sectional age sample of non-human primates (n=39; 22 platyrrhines; 17 strepsirrhines), we used microcomputed tomographic (µCT) and histological methods to track age changes in the sphenoid bone. In the midline, the sphenoid expands its dimensions at three growth centers, including the sphenooccipital, intrasphenoidal (ISS) and presphenoseptal (PSept) synchondroses. Bilaterally, the alisphenoid is enlarged via appositional bone growth that radiates outward from cartilaginous parts of the alisphenoid during midfetal stages. The alisphenoid remains connected to the basitrabecular process of the basisphenoid via the alibasisphenoidal synchondrosis (ABS). Reactivity to proliferating cell-nuclear antigen is observed in all synchondroses, indicating active growth perinatally. Between mid-fetal and birth ages in Saguinus geoffroyi, all synchondroses decrease in the breadth of proliferating columns of chondrocytes. In most primates, the ABS is greatly diminished by birth, and is likely the earliest to fuse, although at least some cartilage may remain by at least one-month of age. Unlike humans, no non-human primate in our sample exhibits perinatal fusion of ISS. A dichotomy among primates is the orientation of the ABS, which is more rostrally directed in platyrrhines. Based on fetal Saguinus geoffroyi specimens, the ABS was initially oriented within a horizontal plane, and redirects inferiorly during late fetal and perinatal stages. These changes occur in tandem with forward orientation of the orbits in platyrrhines, combined with downward growth of the midface. Thus, we postulate that active growth centers direct the orientation of the midface and orbit before birth.

Integration of the sphenoid and ethmoid bones during early postnatal development is poorly described in the literature. A uniquely prolonged patency of sphenoethmoidal synchondrosis, or prespheno-septal synchondrosis (PSept) has been attributed to humans. However, the sphenoethmoidal junction has not been studied using a comparative primate sample. Here, we examined development of the sphenoethmoidal interface using ontogenetic samples of Old and New World monkeys, strepsirrhine primates (lemurs and lorises), and a comparative sample of other mammals. Specimens ranging from late fetal to one month postnatal age were studied using histology, immunohistochemistry, and micro-computed tomography methods. Our results demonstrate that humans are not unique in anterior cranial base growth at PSept, since it is patent in all newborn primates. We found two distinctions within our sample. First, nearly all primates exhibit an earlier breakdown of the nasal capsule cartilage that abuts the orbitosphenoid when compared to non-primates. This may facilitate earlier postnatal integration of the basicranium and midface and may enhance morphological plasticity in the region. Second, the PSept exhibits a basic dichotomy between strepsirrhines and monkeys. In strepsirrhines, the PSept has proliferating chondrocytes that are primarily oriented in a longitudinal plane, as in other mammals. In contrast, monkeys have a convex anterior end of the presphenoid with a radial boundary of cartilaginous growth at PSept. Our findings suggest that the PSept acts as a "pacemaker" of longitudinal facial growth in mammals with relatively long snouts, but may also contribute to facial height and produce a relatively taller midface in anthropoid primates. This article is protected by copyright. All rights reserved.

'Macrosmatic' mammals have dedicated olfactory regions within their nasal cavity and segregated airstreams for olfaction and respiratory air-conditioning. Here, we examined the 3D distribution of olfactory surface area (SA) and nasal airflow patterns in the pygmy slow loris (

Kabuki syndrome 1 (KS1) is a Mendelian disorder of the epigenetic machinery caused by mutations in the gene encoding KMT2D, which methylates lysine 4 on histone H3 (H3K4). KS1 is characterized by intellectual disability, postnatal growth retardation, and distinct craniofacial dysmorphisms. A mouse model (Kmt2d+/ β Geo) exhibits features of the human disorder and has provided insight into other phenotypes; however, the mechanistic basis of skeletal abnormalities and growth retardation remains elusive. Using high-resolution micro-computed tomography we show thatKmt2d+/βGeomice have shortened long bones and ventral bowing of skulls.In vivoexpansion of growth plates within both the skull and long bones suggests disrupted endochondral ossification as a common disease mechanism. Stable chondrocyte cell lines harboring inactivating mutations inKmt2dexhibit increased proliferation and differentiation, which further supports this mechanism. A known inducer of chondrogenesis, SOX9, and its targets show markedly increased expression inKmt2d-/-chondrocytes. By transcriptome profiling, we identifyShox2as a putative KMT2D target. We propose that decreased KMT2D-mediated H3K4me3 atShox2releasesSox9inhibition and thereby leads to enhanced chondrogenesis, providing a novel and plausible explanation for precocious chondrocyte differentiation. Our findings not only provide insight into the pathogenesis of growth retardation in KS1, but also suggest novel therapeutic targets to rescue growth retardation in KS1 and related disorders.

'Macrosmatic' mammals have dedicated olfactory regions within their nasal cavity and segregated airstreams for olfaction and respiratory air-conditioning. Here, we examined the 3D distribution of olfactory surface area (SA) and nasal airflow patterns in the pygmy slow loris (Nycticebus pygmaeus), a primate with primitive nasal cavities, except for enlarged eyes that converge upon the posterodorsal nasal region. Using the head of an adult loris cadaver, we co-registered micro-computed tomography (CT) slices and histology sections to create a 3D reconstruction of the olfactory mucosa distribution. Histological sections were used to measure olfactory surface area and to annotate CT reconstructions. The loris has a complex olfactory recess (∼19% of total nasal SA) with multiple olfactory turbinals. However, the first ethmoturbinal has a rostral projection that extends far anterior to the olfactory recess, lined by ∼90% non-olfactory epithelium. Only one (of three) frontoturbinals bears olfactory mucosa. Computational fluid dynamics simulations of nasal airflow and odorant deposition revealed that there is some segregation of respiratory and olfactory flow in the loris nose, but that it is not as distinct as in well-studied 'macrosmats' (e.g. the dog). In the loris, airflow is segregated medially and laterally to vertically elongated, plate-like first ethmoturbinals. Thus, lorises may be said to have certain macrosmatic anatomical characteristics (e.g. olfactory recess), but not segregated nasal airflow patterns that are optimized for olfaction, as in canids. These results imply that a binary 'microsmatic/macrosmatic' dichotomy does not exist. Rather, mammals appear to exhibit complex trends with respect to specialization of the turbinals and recesses.

We introduce a new method to geometrically reconstruct eye volume and placement in small-bodied primates based on the three-dimensional contour of the intraorbital surface. We validate it using seven species of living primates, with dry skulls and wet dissections, and test its application on seven species of Paleogene fossils of interest. The method performs well even when the orbit is damaged and incomplete, lacking the postorbital bar and represented only by the orbital floor. Eye volume is an important quantity for anatomic and metabolic reasons, which due to differences in eye set, or position within (or outside) the bony orbit, can be underestimated in living and fossil forms when calculated from aperture diameter. Our Ectopic Index quantifies how much the globe's volume protrudes anteriorly from the aperture. Lemur, Notharctus and Rooneyia resemble anthropoids, with deeply recessed eyes protruding 11%-13%. Galago and Tarsius are the other extreme, at 47%-56%. We argue that a laterally oriented aperture has little to do with line-of-sight in euprimates, as large ectopic eyes can position the cornea to enable a directly forward viewing axis, and soft tissue positions the eyes facing forward in megachiropteran bats, which have unenclosed, open eye sockets. The size and set of virtual eyes reconstructed from 3D cranial models confirm that eyes were large to hypertrophic in Hemiacodon, Necrolemur, Microchoerus, Pseudoloris and Shoshonius, but eye size in Rooneyia may have been underestimated by measuring the aperture, as in Aotus. Anat Rec, 299:1671-1689, 2016. © 2016 Wiley Periodicals, Inc.

Living primates have relatively large eyes and support orbital tissues with a postorbital bar (POB) and/or septum. Some mammals with large eyes lack a POB, and presumably rely on soft tissues. Here, we examined the orbits of four species of strepsirrhine primates (Galagidae, Cheirogaleidae) and three species of fruit bats (Pteropodidae). Microdissection and light microscopy were employed to identify support structures of the orbit. In bats and primates, there are two layers of fascial sheets that border the eye laterally. The outer membrane is the most superficial layer of deep fascia, and has connections to the POB in primates. In fruit bats, which lacked a POB or analogous ligament, the deep fascia is reinforced by transverse ligaments. Bats and primates have a deeper membrane supporting the eye, identified as the periorbita (PA) based on the presence of elastic fibers and smooth muscle. The PA merges with periostea deep within the orbit, but has no periosteal attachment to the POB of primates. These findings demonstrate that relatively big eyes can be supported primarily with fibrous connective tissues as well as the PA, in absence of a POB or ligament. The well-developed smooth muscle component within the PA of fruit bats likely helps to protrude the eye, maintaining a more convergent eye orientation, with greater overlap of the visual fields. The possibility should be considered that early euprimates, and even stem primates that may have lacked a POB, also had more convergent eyes than indicated by osseous measurements of orbital orientation. Anat Rec, 299:1690-1703, 2016. © 2016 Wiley Periodicals, Inc.

Bony structure of the postorbital region is a key trait distinguishing major clades of primates. Strepsirrhines share a postorbital bar, and anthropoids share a complete postorbital septum. At issue is whether the partial postorbital septum of tarsiers unites living tarsiers more closely with anthropoids than with certain large-eyed Eocene fossils. Previously we reported incomplete postorbital closure in tarsiers at birth. In this article, we document comparative analyses of the postorbital region in a broad range of perinatal primates. Virtual reconstructions of microCT data were used to study three-dimensional structure of the perinatal cranium in these taxa. We also describe and illustrate formation of the tarsier partial postorbital septum through the perinatal period using a growth series of Tarsius syrichta. Our results support the hypothesis that partial postorbital septation in the tarsier is secondary to eye hypertrophy. Based on these observations, we propose a structural hypothesis for phylogenetic differences observed in the primate postorbital region. Specifically, we propose that key postorbital traits, including the frontal spur in strepsirrhines and the posterior lamina of the zygomatic in anthropoids, develop as a result of the spatial relationships of brain, eyes, and teeth. Haplorhines are united by expansion of the anterior cranial fossa and loss of the frontal spur. Anthropoids are further united to the exclusion of tarsiers by expansion of the temporal lobes and associated formation of the posterior lamina of the zygomatic. Mechanical forces related to these spatial relationships may be modulated by deep fascia of the orbit to induce formation of the postorbital septum. Anat Rec, 299:1631-1645, 2016. © 2016 Wiley Periodicals, Inc.

The nasal cavity of strepsirrhine primates (lemurs and lorises) has the most primitive arrangement of extant primates. In nocturnal species, the numerous turbinals of the ethmoid bear a large surface area of olfactory mucosa (OM). In this study, we examine turbinal development in four genera of diurnal or cathemeral lemuriformes. In addition, we examined an age series of each genus to detect whether structures bearing OM as opposed to respiratory mucosa (RM) develop differently, as has been observed in nocturnal strepsirrhines. In adults, the maxilloturbinal is covered by highly vascular respiratory mucosa throughout its entire length, with large sinusoidal vessels in the lamina propria; any parts of other turbinals that closely borders the maxilloturbinal has a similar mucosa. Posteriorly, the most vascular RM is restricted in the nasopharyngeal duct, which becomes partitioned from the dorsal olfactory region. A comparison of newborns to adults reveals that the first ethmoturbinal increases more in length in the parts that are covered with RM than OM, which supports the idea that ethmoturbinals can specialize in more than one function. Finally, we observe that the regions of turbinals that are ultimately covered with RM develop more accessory lamellae or additional surface area of existing scrolls compared to the regions covered with OM. Because such outgrowths of bone develop postnatally and without cartilaginous precursors, we hypothesize that the complexity of olfactory lamellae within the ethmoturbinal complex is primarily established at birth, while respiratory lamellae become elaborated due to the epigenetic influence of respiratory physiology. Anat Rec, 299:1492-1510, 2016. © 2016 Wiley Periodicals, Inc.

Through artificial selection, humans have altered the morphology of domestic dogs and created the range of morphological traits present in the diverse dog breeds seen today. This article tests how artificial breeding for extreme facial reduction affects the craniodental complex in domestic dogs (Canis lupus familiaris). These brachycephalic dog breeds allow for a natural experiment on facial reduction, providing additional insight into this well-studied phenomenon associated with the evolutionary history of other animals including primates. We used an existing set of clinically obtained CT scans from dogs representing three brachycephalic breeds and a comparative sample. We recorded three-dimensional coordinate data for 62 cranial and mandibular landmarks that allowed us to analyze differences in shape in the neurocranium, basicranium, and viscerocranium. Through geometric morphometric analysis, we confirmed that most of the cranial shape variance in our sample was associated with cephalic index (cranial width-to-length ratio) and reflects the effects of facial reduction. Shape changes in the cranium and mandible co-vary. We also identified several areas of localized shape change. Whereas the primary shape change seen in the mandible is a uniform shift in proportion involving rostral-caudal shortening and medial-lateral widening, the majority of the shape change in the palate is localized to the maxilla. Here, a pronounced difference in maxillary carnassial orientation relative to the mandibular carnassials results in disruption of the functionally important carnassial complex. These results support previous studies showing integration within the skull and highlight the deleterious effects of artificial selection for extreme facial reduction in domestic dogs. Anat Rec, 2019. © 2019 Wiley Periodicals, Inc. Anat Rec, 303:330-346, 2020. © 2019 American Association for Anatomy.