Publications

2014

Brien, Haley D O, and Susan H Williams. 2014. “Using Biplanar Fluoroscopy to Guide Radiopaque Vascular Injections: A New Method for Vascular Imaging”. PloS One 9 (5): e97940. https://doi.org/10.1371/journal.pone.0097940.
Publisher's Version
Studying vascular anatomy, especially in the context of relationships with hard tissues, is of great interest to biologists. Vascular studies have provided significant insight into physiology, function, phylogenetic relationships, and evolutionary patterns. Injection of resin or latex into the vascular system has been a standard technique for decades. There has been a recent surge in popularity of more modern methods, especially radiopaque latex vascular injection followed by CT scanning and digital "dissection." This technique best displays both blood vessels and bone, and allows injections to be performed on cadaveric specimens. Vascular injection is risky, however, because it is not a standardizable technique, as each specimen is variable with regard to injection pressure and timing. Moreover, it is not possible to view the perfusion of injection medium throughout the vascular system of interest. Both data and rare specimens can therefore be lost due to poor or excessive perfusion. Here, we use biplanar video fluoroscopy as a technique to guide craniovascular radiopaque latex injection. Cadaveric domestic pigs (Sus scrofa domestica) and white-tailed deer (Odocoileus virginianus) were injected with radiopaque latex under guidance of fluoroscopy. This method was found to enable adjustments, in real-time, to the rate, location, and pressure at which latex is injected in order to avoid data and specimen loss. In addition to visualizing the injection process, this technique can be used to determine flow patterns, and has facilitated the development of consistent markers for complete perfusion.
Thompson, Cynthia L, Susan H Williams, Kenneth E Glander, Mark F Teaford, and Christopher J Vinyard. (2014) 2014. “Body Temperature and Thermal Environment in a Generalized Arboreal Anthropoid, Wild Mantled Howling Monkeys (Alouatta Palliata).”. American Journal of Physical Anthropology 154 (1): 1-10. https://doi.org/10.1002/ajpa.22505.
Publisher's Version

Free-ranging primates are confronted with the challenge of maintaining an optimal range of body temperatures within a thermally dynamic environment that changes daily, seasonally, and annually. While many laboratory studies have been conducted on primate thermoregulation, we know comparatively little about the thermal pressures primates face in their natural, evolutionarily relevant environment. Such knowledge is critical to understanding the evolution of thermal adaptations in primates and for comparative evaluation of humans' unique thermal adaptations. We examined temperature and thermal environment in free-ranging, mantled howling monkeys (Alouatta palliata) in a tropical dry forest in Guanacaste, Costa Rica. We recorded subcutaneous (Tsc ) and near-animal ambient temperatures (Ta ) from 11 animals over 1586.5 sample hours during wet and dry seasons. Howlers displayed considerable variation in Tsc , which was largely attributable to circadian effects. Despite significant seasonal changes in the ambient thermal environment, howlers showed relatively little evidence for seasonal changes in Tsc . Howlers experienced warm thermal conditions which led to body cooling relative to the environment, and plateaus in Tsc at increasingly warm Ta . They also frequently faced cool thermal conditions (Ta  < Tsc ) in which Tsc was markedly elevated compared with Ta . These data add to a growing body of evidence that non-human primates have more labile body temperatures than humans. Our data additionally support a hypothesis that, despite inhabiting a dry tropical environment, howling monkeys experience both warm and cool thermal pressures. This suggests that thermal challenges may be more prevalent for primates than previously thought, even for species living in nonextreme thermal environments.

O’Brien, Haley D, and Susan H Williams. (2014) 2014. “Using Biplanar Fluoroscopy to Guide Radiopaque Vascular Injections: A New Method for Vascular Imaging.”. PloS One 9 (5): e97940. https://doi.org/10.1371/journal.pone.0097940.
Publisher's Version

Studying vascular anatomy, especially in the context of relationships with hard tissues, is of great interest to biologists. Vascular studies have provided significant insight into physiology, function, phylogenetic relationships, and evolutionary patterns. Injection of resin or latex into the vascular system has been a standard technique for decades. There has been a recent surge in popularity of more modern methods, especially radiopaque latex vascular injection followed by CT scanning and digital "dissection." This technique best displays both blood vessels and bone, and allows injections to be performed on cadaveric specimens. Vascular injection is risky, however, because it is not a standardizable technique, as each specimen is variable with regard to injection pressure and timing. Moreover, it is not possible to view the perfusion of injection medium throughout the vascular system of interest. Both data and rare specimens can therefore be lost due to poor or excessive perfusion. Here, we use biplanar video fluoroscopy as a technique to guide craniovascular radiopaque latex injection. Cadaveric domestic pigs (Sus scrofa domestica) and white-tailed deer (Odocoileus virginianus) were injected with radiopaque latex under guidance of fluoroscopy. This method was found to enable adjustments, in real-time, to the rate, location, and pressure at which latex is injected in order to avoid data and specimen loss. In addition to visualizing the injection process, this technique can be used to determine flow patterns, and has facilitated the development of consistent markers for complete perfusion.

2013

Chen, Yong, Susan H Williams, Amy L McNulty, Ji Hee Hong, Suk Hee Lee, Nicole E Rothfusz, Puja K Parekh, et al. (2025) 2013. “Temporomandibular Joint Pain: A Critical Role for Trpv4 in the Trigeminal Ganglion”. Pain 154 (8): 1295—1304. https://doi.org/10.1016/j.pain.2013.04.004.
Publisher's Version
Temporomandibular joint disorder (TMJD) is known for its mastication-associated pain. TMJD is medically relevant because of its prevalence, severity, chronicity, the therapy-refractoriness of its pain, and its largely elusive pathogenesis. Against this background, we sought to investigate the pathogenetic contributions of the calcium-permeable TRPV4 ion channel, robustly expressed in the trigeminal ganglion sensory neurons, to TMJ inflammation and pain behavior. We demonstrate here that TRPV4 is critical for TMJ-inflammation-evoked pain behavior in mice and that trigeminal ganglion pronociceptive changes are TRPV4-dependent. As a quantitative metric, bite force was recorded as evidence of masticatory sensitization, in keeping with human translational studies. In Trpv4(-/-) mice with TMJ inflammation, attenuation of bite force was significantly less than in wildtype (WT) mice. Similar effects were seen with systemic application of a specific TRPV4 inhibitor. TMJ inflammation and mandibular bony changes were apparent after injections of complete Freund adjuvant but were remarkably independent of the Trpv4 genotype. It was intriguing that, as a result of TMJ inflammation, WT mice exhibited significant upregulation of TRPV4 and phosphorylated extracellular-signal-regulated kinase (ERK) in TMJ-innervating trigeminal sensory neurons, which were absent in Trpv4(-/-) mice. Mice with genetically-impaired MEK/ERK phosphorylation in neurons showed resistance to reduction of bite force similar to that of Trpv4(-/-) mice. Thus, TRPV4 is necessary for masticatory sensitization in TMJ inflammation and probably functions upstream of MEK/ERK phosphorylation in trigeminal ganglion sensory neurons in vivo. TRPV4 therefore represents a novel pronociceptive target in TMJ inflammation and should be considered a target of interest in human TMJD.

2011

Konow, Nicolai, Anthony Herrel, Callum F Ross, Susan H Williams, Rebecca Z German, Christopher P J Sanford, and Chris Gintof. (2025) 2011. “Evolution of Muscle Activity Patterns Driving Motions of the Jaw and Hyoid During Chewing in Gnathostomes”. Integrative and Comparative Biology 51 (2): 235—246. https://doi.org/10.1093/icb/icr040.
Publisher's Version
Although chewing has been suggested to be a basal gnathostome trait retained in most major vertebrate lineages, it has not been studied broadly and comparatively across vertebrates. To redress this imbalance, we recorded EMG from muscles powering anteroposterior movement of the hyoid, and dorsoventral movement of the mandibular jaw during chewing. We compared muscle activity patterns (MAP) during chewing in jawed vertebrate taxa belonging to unrelated groups of basal bony fishes and artiodactyl mammals. Our aim was to outline the evolution of coordination in MAP. Comparisons of activity in muscles of the jaw and hyoid that power chewing in closely related artiodactyls using cross-correlation analyses identified reorganizations of jaw and hyoid MAP between herbivores and omnivores. EMG data from basal bony fishes revealed a tighter coordination of jaw and hyoid MAP during chewing than seen in artiodactyls. Across this broad phylogenetic range, there have been major structural reorganizations, including a reduction of the bony hyoid suspension, which is robust in fishes, to the acquisition in a mammalian ancestor of a muscle sling suspending the hyoid. These changes appear to be reflected in a shift in chewing MAP that occurred in an unidentified anamniote stem-lineage. This shift matches observations that, when compared with fishes, the pattern of hyoid motion in tetrapods is reversed and also time-shifted relative to the pattern of jaw movement.
Williams, Susan H, Kristin K Stover, Jillian S Davis, and Stephane J Montuelle. (2025) 2011. “Mandibular Corpus Bone Strains During Mastication in Goats (Capra Hircus): A Comparison of Ingestive and Rumination Chewing”. Archives of Oral Biology 56 (10): 960—971. https://doi.org/10.1016/j.archoralbio.2011.02.014.
Publisher's Version

Objective

To compare the mechanical loading environment of the jaw in goats during ingestive and rumination chewing.

Design

Rosette strain gauges were attached to the external surface of the mandibular corpus in five goats to record bone strains during the mastication of hay and rumination.

Results

Strain magnitudes and maximum physiological strain rates during the mastication of hay are significantly higher than during rumination chewing on the working and balancing sides. Principal strain ratios and orientations are similar between the two chewing behaviours. Loading and chewing cycle duration are all longer during rumination chewing, whereas chew duty factor and variances in load and chewing cycle durations are higher during ingestive chewing. For most of the variables, differences in strain magnitudes or durations are similar at all three gauge sites, suggesting that rumination and ingestive chewing do not differentially influence bone at the three gauge sites.

Conclusions

Despite lower strain magnitudes, the repetitive nature of rumination chewing makes it an important component of the mechanical loading environment of the selenodont artiodactyl jaw. However, similarities in principal strain orientations and ratios indicate that rumination chewing need not be considered as a unique loading behaviour influencing the biomechanics of the selenodont artiodactyl jaw. Differences in loading and chewing cycle durations during rumination and ingestion demonstrate flexibility in adult chewing frequencies. Finally, although the low within-sequence variability in chewing cycle durations supports the hypothesis that mammalian mastication is energetically efficient, chewing during rumination may not be more efficient than during ingestion.
Wall, Christine E, Christopher J Vinyard, Susan H Williams, Vladimir Gapeyev, Xianhua Liu, Hilmar Lapp, and Rebecca Z German. (2025) 2011. “Overview of FEED, the Feeding Experiments End-User Database”. Integrative and Comparative Biology 51 (2): 215—223. https://doi.org/10.1093/icb/icr047.
Publisher's Version
The Feeding Experiments End-user Database (FEED) is a research tool developed by the Mammalian Feeding Working Group at the National Evolutionary Synthesis Center that permits synthetic, evolutionary analyses of the physiology of mammalian feeding. The tasks of the Working Group are to compile physiologic data sets into a uniform digital format stored at a central source, develop a standardized terminology for describing and organizing the data, and carry out a set of novel analyses using FEED. FEED contains raw physiologic data linked to extensive metadata. It serves as an archive for a large number of existing data sets and a repository for future data sets. The metadata are stored as text and images that describe experimental protocols, research subjects, and anatomical information. The metadata incorporate controlled vocabularies to allow consistent use of the terms used to describe and organize the physiologic data. The planned analyses address long-standing questions concerning the phylogenetic distribution of phenotypes involving muscle anatomy and feeding physiology among mammals, the presence and nature of motor pattern conservation in the mammalian feeding muscles, and the extent to which suckling constrains the evolution of feeding behavior in adult mammals. We expect FEED to be a growing digital archive that will facilitate new research into understanding the evolution of feeding anatomy.
Stover, Kristin K, and Susan H Williams. (2025) 2011. “Intraspecific Scaling of Chewing Cycle Duration in Three Species of Domestic Ungulates”. The Journal of Experimental Biology 214 (Pt 1): 104—112. https://doi.org/10.1242/jeb.043646.
Publisher's Version
In mammals, chewing cycle duration (CCD) increases with various measures of size, scaling with body mass(0.13-0.28) and jaw length(0.55). Proposed explanations for these scaling relationships include the allometry of body size, basal metabolic rate and tooth size, on the one hand, and pendular mechanics treating the jaw as a gravity-driven pendulum, on the other. Little is known, however, about the relationship between CCD and size within species. Recent research in dogs demonstrates altogether different scaling exponents and weaker correlations. This research suggests that breed-specific growth rates influence the maturation of the neural networks generating chewing rhythm, which may be altered because of changes in jaw mass during early postnatal growth. Here, we explored the intraspecific scaling of CCD within a sample of adult horses ranging from miniatures to draft breeds and an ontogenetic sample of goats and alpacas from infants to adults. In horses, CCD scales with body mass(0.19) and jaw length(0.57), although in neither case is the correlation significant. In the ontogenetic samples of goats and alpacas, CCD is significantly correlated with body mass, scaling as CCD∝body mass(0.37) in both species. In goats, but not alpacas, CCD is also significantly correlated with jaw length, scaling as jaw length(1.032). As in dogs, the scaling of CCD in horses may reflect the influence of selective breeding on growth trajectories of different breeds, resulting in reduced body and jaw size differences among infants, when CCD is established, compared with adults. However, the allometric scaling of tooth size in horses of different breeds may be a potential influence on the scaling of CCD. The scaling of CCD with body and jaw size in goats, and to a lesser extent in alpacas, also suggests that the development of peripheral masticatory structures such as the teeth and occlusal relations may play a role in changes in CCD during the earliest stages of postnatal ontogeny.
Vinyard, Christopher J, Susan H Williams, Christine E Wall, Alison H Doherty, Alfred W Crompton, and William L Hylander. (2025) 2011. “A Preliminary Analysis of Correlations Between Chewing Motor Patterns and Mandibular Morphology across Mammals”. Integrative and Comparative Biology 51 (2): 260—270. https://doi.org/10.1093/icb/icr066.
Publisher's Version
The establishment of a publicly-accessible repository of physiological data on feeding in mammals, the Feeding Experiments End-user Database (FEED), along with improvements in reconstruction of mammalian phylogeny, significantly improves our ability to address long-standing questions about the evolution of mammalian feeding. In this study, we use comparative phylogenetic methods to examine correlations between jaw robusticity and both the relative recruitment and the relative time of peak activity for the superficial masseter, deep masseter, and temporalis muscles across 19 mammalian species from six orders. We find little evidence for a relationship between jaw robusticity and electromyographic (EMG) activity for either the superficial masseter or temporalis muscles across mammals. We hypothesize that future analyses may identify significant associations between these physiological and morphological variables within subgroups of mammals that share similar diets, feeding behaviors, and/or phylogenetic histories. Alternatively, the relative peak recruitment and timing of the balancing-side (i.e., non-chewing-side) deep masseter muscle (BDM) is significantly negatively correlated with the relative area of the mandibular symphysis across our mammalian sample. This relationship exists despite BDM activity being associated with different loading regimes in the symphyses of primates compared to ungulates, suggesting a basic association between magnitude of symphyseal loads and symphyseal area among these mammals. Because our sample primarily represents mammals that use significant transverse movements during chewing, future research should address whether the correlations between BDM activity and symphyseal morphology characterize all mammals or should be restricted to this "transverse chewing" group. Finally, the significant correlations observed in this study suggest that physiological parameters are an integrated and evolving component of feeding across mammals.
Williams, Susan H, Christopher J Vinyard, Christine E Wall, Alison H Doherty, Alfred W Crompton, and William L Hylander. (2025) 2011. “A Preliminary Analysis of Correlated Evolution in Mammalian Chewing Motor Patterns”. Integrative and Comparative Biology 51 (2): 247—259. https://doi.org/10.1093/icb/icr068.
Publisher's Version
Descriptive and quantitative analyses of electromyograms (EMG) from the jaw adductors during feeding in mammals have demonstrated both similarities and differences among species in chewing motor patterns. These observations have led to a number of hypotheses of the evolution of motor patterns, the most comprehensive of which was proposed by Weijs in 1994. Since then, new data have been collected and additional hypotheses for the evolution of motor patterns have been proposed. Here, we take advantage of these new data and a well-resolved species-level phylogeny for mammals to test for the correlated evolution of specific components of mammalian chewing motor patterns. We focus on the evolution of the coordination of working-side (WS) and balancing-side (BS) jaw adductors (i.e., Weijs Triplets I and II), the evolution of WS and BS muscle recruitment levels, and the evolution of asynchrony between pairs of muscles. We converted existing chewing EMG data into binary traits to incorporate as much data as possible and facilitate robust phylogenetic analyses. We then tested hypotheses of correlated evolution of these traits across our phylogeny using a maximum likelihood method and the Bayesian Markov Chain Monte Carlo method. Both sets of analyses yielded similar results highlighting the evolutionary changes that have occurred across mammals in chewing motor patterns. We find support for the correlated evolution of (1) Triplets I and II, (2) BS deep masseter asynchrony and Triplets I and II, (3) a relative delay in the activity of the BS deep masseter and a decrease in the ratio of WS to BS muscle recruitment levels, and (4) a relative delay in the activity of the BS deep masseter and a delay in the activity of the BS posterior temporalis. In contrast, changes in relative WS and BS activity levels across mammals are not correlated with Triplets I and II. Results from this work can be integrated with dietary and morphological data to better understand how feeding and the masticatory apparatus have evolved across mammals in the context of new masticatory demands.