Non-avian Reptiles Research Articles

Exploring elbow kinematics in the central bearded dragon (Pogona vitticeps) using XROMM: Implications for the role of forearm long-axis rotation in non-avian reptile posture and mobility.

The functional morphology and kinematics of the elbow joint remain relatively understudied in squamates. Previous investigations of lizard elbow morphology and kinematics suggest long-axis rotation (LAR) of the radius and ulna during stance allows the manus to remain pronated during forelimb retraction. Using XROMM (X-ray Reconstruction of Moving Morphology), we explored the range of 3D movements and kinematics of the humerus, radius, and ulna in three adult male Central bearded dragons (Pogona vitticeps) during trackway walking. Our data indicate that the elbow joint of P. vitticeps experiences significant rotations in all three dimensions and that the radius and ulna adduct and rotate laterally on their long axes relative to the elbow joint and to one another during stance. These movements allow the distal ends of the radius and ulna to remain in a configuration necessary for manus pronation. These data support previous inferences that the radius and ulna of lizards move independently at the wrist joint. We suggest that independent LAR of the radius and ulna relative to the elbow joint and to one another may be an ancestral mechanism in lizards and perhaps more broadly across non-avian reptiles.

Palaeognaths Reveal Evolutionary Ancestry of the Avian Major Histocompatibility Complex Class II.

The multigene family of the major histocompatibility complex (MHC) codes for the key antigen-presenting molecules of the vertebrate immune system. In birds, duplicated MHC class II (MHC-II) genes are highly homogenized by concerted evolution, and thus, identification of their orthologous relationships across long evolutionary timescales remains challenging. Relatively low evolutionary rate of avian MHC class IIA genes has been expected to provide a promising avenue to allow such inferences, but availability of MHC-IIA sequences in nonmodel bird species has been limited until recently. Here, taking advantage from accumulating genomic resources, we identified and analyzed MHC-IIA sequences from the most basal lineage of extant birds (Palaeognathae). Conserved region of the MHC-IIA membrane-proximal domain was used to search for orthologous relationships between palaeognath birds and nonavian reptiles. First, analyses of palaeognath sequences revealed the presence of a separate MHC-IIA gene lineage (DAA3) in kiwis, which did not cluster with previously described avian MHC-IIA lineages (DAA1 and DAA2). Next, phylogenetic reconstruction showed that kiwi DAA3 sequences form a single well-supported cluster with turtle MHC-IIA. High similarity of these sequences most likely reflects their remarkable evolutionary conservation and retention of ancient orthologous relationships, which can be traced back to basal archosauromorphs ca. 250 million years ago. Our analyses offer novel insights into macroevolutionary history of the MHC and reinforce the view that rapid accumulation of high-quality genome assemblies across divergent nonmodel species can substantially advance our understanding of gene evolution.

On the function and origin of the avian renal portal shunt and its potential significance throughout evolution.

All birds possess a unique venous architecture surrounding the kidneys known as the renal portal system. In veterinary medicine, this system is well known for causing a first-pass effect when medication is administered parenterally via the leg veins, that is venous blood from the leg is filtered before entering general circulation, thus possibly compromising adequate dosage. Additionally, bilateral valves are present in these veins, and it has been hypothesized that they play a crucial role in regulating flow through the kidneys to protect them against increases in blood pressure. While this hypothesis has been acknowledged, it has not been thoroughly explored. We propose that the function of the renal portal valve extends beyond its significance for kidney function, potentially impacting general hemodynamics. Examining anatomical similarities with extant non-avian reptiles, which lack the renal portal shunt with valve, could reveal additional functionalities of this system in birds. Given the endothermic metabolism and the energetically expensive locomotor activity of birds, the resistance of the hepatic and renal portal system might constrain the blood flow from splanchnic to non-splanchnic blood vessels necessary for (sustained) peak performance. Therefore, diverting blood from the renal portal system using the renal portal valve as a regulatory structure might represent a key adaptation to facilitate sustained peak performance. In addition, we hypothesize that this shunt and valve represents a very early adaptation in amniotes, possibly lost in extant non-avian reptiles but enhanced in birds, with a pivotal role in maintaining hemodynamic homeostasis to support the high metabolic rates characteristic of birds.

Bioimaging of sense organs and the central nervous system in extant fishes and reptiles in situ: A review.

Bioimaging is changing the field of sensory biology, especially for taxa that are lesser-known, rare, and logistically difficult to source. When integrated with traditional neurobiological approaches, developing an archival, digital repository of morphological images can offer the opportunity to improve our understanding of whole neural systems without the issues of surgical intervention and negate the risk of damage and artefactual interpretation. This review focuses on current approaches to bioimaging the peripheral (sense organs) and central (brain) nervous systems in extant fishes (cartilaginous and bony) and non-avian reptiles in situ. Magnetic resonance imaging (MRI), micro-computed tomography (μCT), both super-resolution track density imaging and diffusion tensor-based imaging, and a range of other new technological advances are presented, together with novel approaches in optimizing both contrast and resolution, for developing detailed neuroanatomical atlases and enhancing comparative analyses of museum specimens. For MRI, tissue preparation, including choice of fixative, impacts tissue MR responses, where both resolving power and signal-to-noise ratio improve as field strength increases. Time in fixative, concentration of contrast agent, and duration of immersion in the contrast agent can also significantly affect relaxation times, and thus image quality. For μCT, the use of contrast-enhancing stains (iodine-, non-iodine-, or nanoparticle-based) is critical, where the type of fixative used, and the concentration of stain and duration of staining time often require species-specific optimization. Advanced reconstruction algorithms to reduce noise and artifacts and post-processing techniques, such as deconvolution and filtering, are now being used to improve image quality and resolution.

Gekko gecko as a model organism for understanding aspects of laryngeal vocal evolution.

The ability to communicate through vocalization plays a key role in the survival of animals across all vertebrate groups. Although avian reptiles have received much attention relating to their stunning sound repertoire, non-avian reptiles have been wrongfully assumed to have less elaborate vocalization types, and little is known about the biomechanics of sound production and their underlying neural pathways in this group. We investigated alarm calls of Gekko gecko using audio and cineradiographic recordings. Acoustic analysis revealed three distinct call types: a sinusoidal call type (type 1); a train-like call type, characterized by distinct pulse trains (type 3); and an intermediate type, which showed both sinusoidal and pulse train components (type 2). Kinematic analysis of cineradiographic recordings showed that laryngeal movements differ significantly between respiratory and vocal behavior. During respiration, animals repeatedly moved their jaws to partially open their mouths, which was accompanied by small glottal movements. During vocalization, the glottis was pulled back, contrasting with what has previously been reported. In vitro retrograde tracing of the nerve innervating the laryngeal constrictor and dilator muscles revealed round to fusiform motoneurons in the hindbrain-spinal cord transition ipsilateral to the labeled nerve. Taken together, our observations provide insight into the alarm calls generated by G. gecko, the biomechanics of this sound generation and the underlying organization of motoneurons involved in the generation of vocalizations. Our observations suggest that G. gecko may be an excellent non-avian reptile model organism for enhancing our understanding of the evolution of vertebrate vocalization.

Karyotype depends on sperm head morphology in some amniote groups.

The karyotype of an organism is the set of gross features that characterize the way the genome is packaged into separate chromosomes. It has been known for decades that different taxonomic groups often have distinct karyotypic features, but whether selective forces act to maintain these differences over evolutionary timescales is an open question. In this paper we analyze a database of karyotype features and sperm head morphology in 103 mammal species with spatulate sperm heads and 90 sauropsid species (birds and non-avian reptiles) with vermiform heads. We find that mammal species with a larger head area have more chromosomes, while sauropsid species with longer heads have a wider range of chromosome lengths. These results remain significant after controlling for genome size, so sperm head morphology is the relevant variable. This suggest that post-copulatory sexual selection, by acting on sperm head shape, can influence genome architecture.

Hidden social complexity behind vocal and acoustic communication in non-avian reptiles.

Social interactions are inevitable in the lives of most animals, since most essential behaviours require interaction with conspecifics, such as mating and competing for resources. Non-avian reptiles are typically viewed as solitary animals that predominantly use their vision and olfaction to communicate with conspecifics. Nevertheless, in recent years, evidence is mounting that some reptiles can produce sounds and have the potential for acoustic communication. Reptiles that can produce sound have an additional communicative channel (in addition to visual/olfactory channels), which could suggest they have a higher communicative complexity, the evolution of which is assumed to be driven by the need of social interactions. Thus, acoustic reptiles may provide an opportunity to unveil the true social complexity of reptiles that are usually thought of as solitary. This review aims to reveal the hidden social interactions behind the use of sounds in non-avian reptiles. Our review suggests that the potential of vocal and acoustic communication and the complexity of social interactions may be underestimated in non-avian reptiles, and that acoustic reptiles may provide a great opportunity to uncover the coevolution between sociality and communication in non-avian reptiles. This article is part of the theme issue 'The power of sound: unravelling how acoustic communication shapes group dynamics'.

Hydric Environment and Chemical Composition Shape Non-avian Reptile Eggshell Absorption.

The amniotic egg fulfils a critical role in reproduction by serving as an interface between the external environment and the embryo. Because non-avian reptiles are rarely incubated, they must be heated by, and absorb water from, the oviposition site for the developing embryo. The mechanisms by which they absorb sufficient, but not excess, water and how these mechanisms vary with local habitat is largely unknown, despite its significance to their evolution. Here, we first performed histology, Fourier-transform infrared spectroscopy and dynamic vapor sorption experiments to elucidate the mechanisms of eggshell absorption for 56 reptile species. Then, we used phylogenetic comparative analysis to test the hypothesis that the absorptive capacity of reptile eggshells increases with aridity of the environment. We found that water absorption increases in the presence of a superficial mucopolysaccharide layer and decreases with increased calcium content. We found that eggs from arid environments have highly absorbent eggshells, but only in species with weakly calcified shells. Our results suggest that reptile eggshells have over evolutionary time tuned absorptive capacity to environmental moisture level. Since these eggs often must sustain conflicting constraints, they may serve as inspirations for new biomimetic materials, such as water filtering membranes or humidity sensors.

Structural analysis of the female reptile reproductive system by micro-computed tomography and optical coherence tomography†.

Volumetric data provide unprecedented structural insight to the reproductive tract and add vital anatomical context to the relationships between organs. The morphology of the female reproductive tract in non-avian reptiles varies between species, corresponding to a broad range of reproductive modes and providing valuable insight to comparative investigations of reproductive anatomy. However, reproductive studies in reptilian models, such as the brown anole studied here, have historically relied on histological methods to understand the anatomy. While these methods are highly effective for characterizing the cell types present in each organ, histological methods lose the 3D relationships between images and leave the architecture of the organ system poorly understood. We present the first comprehensive volumetric analyses of the female brown anole reproductive tract using two non-invasive, non-destructive imaging modalities: micro-computed tomography (microCT) and optical coherence tomography (OCT). Both are specialized imaging technologies that facilitate high-throughput imaging and preserve three-dimensional information. This study represents the first time that microCT has been used to study all reproductive organs in this species and the very first time that OCT has been applied to this species. We show how the non-destructive volumetric imaging provided by each modality reveals anatomical context including orientation and relationships between reproductive organs of the anole lizard. In addition to broad patterns of morphology, both imaging modalities provide the high resolution necessary to capture details and key anatomical features of each organ. We demonstrate that classic histological features can be appreciated within whole-organ architecture in volumetric imaging using microCT and OCT, providing the complementary information necessary to understand the relationships between tissues and organs in the reproductive system. This side-by-side imaging analysis using microCT and OCT allows us to evaluate the specific advantages and limitations of these two methods for the female reptile reproductive system.

Dawn of snake sociality: resource defence and social behaviours of Lycodon semicarinatus foraging on sea turtles

Abstract Snakes are traditionally considered to be the least social among non-avian reptiles. Therefore, social behaviours that are not related to mating have rarely been studied in wild snakes. Nonetheless, in two species of snakes, Lycodon semicarinatus and Oligodon formosanus, intraspecific interactions unrelated to mating have been reported among individuals that forage on sea turtles and cluster around sea turtle nests. In this study, we conducted a field survey on L. semicarinatus on a sandy beach on Okinawa Island for four years by route census and fixed videography to quantify interactions between individuals that occurred at sea turtle nests. Snakes repelled conspecifics from the nest by using a ritual combat dance. Whether the snakes attacked the conspecific depended on the opponent at the nest. Some snakes waited for the conspecific to leave the nest. This decision-making may be based on individual recognition by chemical cues, because the snakes exhibited frequent tongue-flicking toward the occupant before contacting. Their tongue-flicking duration was significantly longer when the subsequent response was attacking than when it was fleeing. These intraspecific interactions imply the occurrence of territoriality and suggest the possible occurrence of a dominance hierarchy in L. semicarinatus. A comparison of L. semicarinatus with a previous study on O. formosanus revealed that they express different social interactions, although the two species use a similar feeding environment. Oligodon formosanus removes conspecifics from the nest by simply biting, but L. semicarinatus competed for the priority to access the nest by combat dance, and its decision-making whether to challenge the combat dance changed flexibly depending on the opponent. In addition, several L. semicarinatus that did not challenge the combat dance waited for hours near the opponent that were foraging on the nest. Lycodon semicarinatus exhibited behaviours to avoid encounter and fights with conspecific individuals. Although there are differences in the social level between the two snake species, the exploitation of a food resource that contains large numbers of prey in one place, such as sea turtle nests, may be an ecological factor that favours the evolutionary development of social behaviours in solitary snakes.

Developmental energetics in the oviparous corn snake, Pantherophis guttatus, confirms a conservative evolutionary pattern in snakes

Eggs of oviparous reptiles are ideal models for studying evolutionary patterns of embryonic metabolism since they allow tracking of energy allocation during development. Analyzing oxygen consumption of whole eggs throughout development indicates three patterns among reptiles. Embryos initially grow and consume oxygen exponentially, but oxygen consumption slows, or drops before hatching in some species. Turtles, crocodilians, and most lizards follow curves with initial exponential increases followed by declines, whereas embryonic snakes that have been studied exhibit a consistently exponential pattern. This study measured oxygen consumption of corn snake, Pantherophis guttatus, embryos to determine if this species also exhibits an exponential increase in oxygen consumption. Individual eggs, sampled weekly from oviposition to hatching, were placed in respirometry chambers for 24-h during which oxygen consumption was recorded. Embryos were staged and carcasses and yolk were weighed separately. Results indicate steady inclines in oxygen consumption during early stages of development, with a rapid increase prior to hatching. The findings support the hypothesis that embryonic oxygen consumption of snakes differs from most other non-avian reptiles. Total energy required for development was determined based on calorimetry of initial yolk compared to hatchlings and residual yolk and by integration of the area under the curve plotting oxygen consumption versus age of embryos. The cost of development estimates based on these two methods were 6.4 and 10.0 kJ, respectively. Our results emphasize the unique physiological aspects of snake embryogenesis and illustrate how the study of physiological characteristics can contribute to the broader understanding of reptilian evolution.

A review of age estimation methods in non-avian reptiles by growth marks in hard tissues.

Age and growth-related data are basic biological parameters, essential in population ecology, evolution, and conservation biology. There is a growing body of published information on reptile demography derived from sclerochronology, a technique based on counting the growth layers deposited in bones (skeletochronology) and other hard body structures. Since the data are not always easily available, we compiled the existing published data, described the current status of knowledge, synthetized the conclusions of disparate studies, and identified patterns of research and information gaps, prioritizing the needs for future research. Our database includes the results of 468 published studies covering 236 reptile species from 41 families. These represent less than 2% of the total number of known extant species. Turtles and crocodiles are proportionally better studied, while snakes are the least examined group. The distribution of the research does not reflect conservation needs; we found an important geographic bias, with an overrepresentation of Northern temperate species. Only 23% of the studies checked the assumption of periodicity of growth marks deposition, and the method was found to be reliable or adequate in 79% of the cases. Overall, the data obtained through sclerochronology can be considered robust, especially if validation methods are employed, since the general goal is to characterize population parameters, trends, and dynamics, rather than determining the exact age of any specimen in particular.

Does heat tolerance actually predict animals' geographic thermal limits?

The “climate extremes hypothesis” is a major assumption of geographic studies of heat tolerance and climatic vulnerability. However, this assumption remains vastly untested across taxa, and multiple factors may contribute to uncoupling heat tolerance estimates and geographic limits. Our dataset includes 1000 entries of heat tolerance data and maximum temperatures for each species' known geographic limits (hereafter, Tmax). We gathered this information across major animal taxa, including marine fish, terrestrial arthropods, amphibians, non-avian reptiles, birds, and mammals. We first tested if heat tolerance constrains the Tmax of sites where species could be observed. Secondly, we tested if the strength of such restrictions depends on how high Tmax is relative to heat tolerance. Thirdly, we correlated the different estimates of Tmax among them and across species. Restrictions are strong for amphibians, arthropods, and birds but often weak or inconsistent for reptiles and mammals. Marine fish describe a non-linear relationship that contrasts with terrestrial groups. Traditional heat tolerance measures in thermal vulnerability studies, like panting temperatures and the upper set point of preferred temperatures, do not predict Tmax or are inversely correlated to it, respectively. Heat tolerance restricts the geographic warm edges more strongly for species that reach sites with higher Tmax for their heat tolerance. These emerging patterns underline the importance of reliable species' heat tolerance indexes to identify their thermal vulnerability at their warm range edges. Besides, the tight correlations of Tmax estimates across on-land microhabitats support a view of multiple types of thermal challenges simultaneously shaping ranges' warm edges for on-land species. The heterogeneous correlation of Tmax estimates in the ocean supports the view that fish thermoregulation is generally limited, too. We propose new hypotheses to understand thermal restrictions on animal distribution.

Female long-term sperm storage results in viable offspring in the Himalayan Mountain Pitviper, Ovophis monticola.

The ability of females to store sperm for extended periods in their reproductive tracts (termed long-term sperm storage, LTSS) has been reported across a diversity of vertebrate taxa. The evolutionary, ecological, and physiological significance of LTSS is wide-ranging and includes the ability to produce offspring when mates may be temporally scarce by way of decoupling copulation from ovulation, inbreeding avoidance, and the generation and maintenance of genetic diversity in progeny. Among vertebrate lineages, nonavian reptiles exhibit a remarkable capacity for LTSS, with the production of viable offspring reported after periods exceeding 6 years since prior contact with a potential mate. Given that female reptiles are able to store viable sperm for prolonged periods, it is important to disentangle LTSS from that of facultative parthenogenesis (FP), a reproductive trait which appears widespread among all reptile lineages. The implications of this distinction are particularly important in the context of the development and management of captive breeding programs. To accurately determine between the two reproductive strategies, genomic screening is highly recommended. Following a period of isolation for 13 months from a potential male mate, a female Himalayan Mountain Pitviper (Ovophis monticola) produced a clutch of three male offspring. Here, through genome-scale analyses of the female and her progeny, we document the first record of LTSS in this genus and exclude FP as the alternative hypothesis.

First successful artificial insemination of a reptile using frozen-thawed semen.

The use of assisted reproductive technologies (ARTs) has considerable value for conservation. However, their development has been limited to relatively few species of charismatic megafauna. Non-avian reptiles are incredibly speciose and up to a third are threatened with extinction, but progress developing and implementing ARTs into conservation programs has lagged other taxonomic groups. Here, we describe successful artificial insemination of an endangered snake using frozen-thawed semen within the context of a species recovery program. Six female Louisiana pinesnakes (Pituophis ruthveni) were inseminated with frozen-thawed semen from six donor males resulting in oviposition of two fertile cutches producing three viable hatchlings. Paternity of hatchlings was confirmed using genetic analyses. This is the first documented case of successful artificial insemination using frozen-thawed semen in a reptile and is a significant step forward in the conservation of reptiles. The methods described here hold the potential for zoos and conservation organizations to diversify reptile gene pools in captivity and the transfer of genetic material between wild populations and captive breeding programs. In addition, the ability to use frozen-thawed semen for AI emphasizes the conservation value of establishing cryobanks for reptile semen. The development and implementation of ARTs in recovery programs for reptiles has great conservation potential and warrants exploration across a broader range of species.

Historical perspective and state of knowledge of the nonavian reptiles (Vertebrata: Sauropsida) of the Department of Caldas, Central Andes of Colombia

Studying non-avian reptiles in the Colombian Andean region has encompassed various spatial scales and historical times. The knowledge of non-avian reptile diversity in the Department of Caldas (Central Andes; 7,888 sq. km. area; intricate geomorphology) has grown significantly in recent decades, especially in the areas to the east of the department within the Magdalena Valley and near Manizales, the capital city. Here, we consolidate the data from biological collections, literature, and unpublished research in a historical synopsis of the studies conducted in Caldas with an updated account of non-avian reptile species recorded in them. We found that the records of non-avian reptiles in Caldas trace back to pre-Columbian and colonial times, with specimens in biological collections dating back over a century. This non-avian reptile richness comprises 126 species grouped in 69 genera, 25 families, and three orders. Three out of the 27 municipalities in Caldas (La Merced, Marulanda, and Marquetalia) lack validated specimens or records. Furthermore, there is limited information regarding protected areas such as Los Nevados and Selva de Florencia National Natural Parks, which demands further research to unravel the diversity patterns of this biological group within this specific segment of the Andes.

A METHOD FOR ISOLATING AND SEQUENCING TRYPANOSOME CELLS TO INVESTIGATE SPECIES ASSOCIATIONS IN MULTIPLE MORPHOTYPE INFECTIONS.

Trypanosome infections containing multiple morphologies have been described from all classes of vertebrates, including mammals, birds, non-avian reptiles, amphibians, and fish. These mixed infections make it challenging to evaluate trypanosome diversity, as it is not immediately clear whether the forms present in the bloodstream represent different species or a single pleomorphic species. Amphibians are common hosts for trypanosomes and are often infected by multiple trypanosome morphologies in the bloodstream. Based on morphological observations and life cycle studies, many authors have considered multiple trypanosome morphotypes found infecting the same frogs to be a single pleomorphic species. However, molecular evidence supporting pleomorphic trypanosome species in amphibians is lacking, primarily because linking sequence data to bloodstream trypanosome morphology in mixed infections is extremely challenging. Here we present methods to isolate individual trypanosome cells of 6 morphotypes from frog blood for nested PCR of the 18S rRNA and gGAPDH genes. Single trypanosome cells were isolated by dilution and 3 DNA extraction methods, and 5 nested PCR primer regimes were utilized to optimize amplification from very low starting concentrations. The success rates of extraction methods ranged from 29 to 50% with the use of a Direct PCR kit having the highest success rate. Although the success rate varied in the different combinations of extraction methods and primer regimes, multiple individuals of all 6 trypanosome morphotypes were sequenced for both genes in a novel way that links sequence data to cell morphology by observing isolated cells with a microscope before PCR amplification. All 6 morphologically distinguishable morphotypes coinfecting a frog were genetically distinct. The only other recent molecular study on amphibian trypanosomes also found genetic differences between morphotypes in multiple infections. Together these studies suggest that the occurrence of pleomorphism may be overestimated in amphibian trypanosomes. The methods presented here offer a promising solution to characterize trypanosome diversity within multiple morphotype infections.

Effects of testosterone on urogenital tract morphology and androgen receptor expression in immature Eastern Fence lizards (Sceloporus undulatus)

In non-avian reptiles, the onset of sexual dimorphism of the major structures of the urogenital tract varies temporally relative to gonadal differentiation, more so than in other amniote lineages. In the current study, we used tonic-release implants to investigate the effects of exogenous testosterone (T) on postnatal development of the urogenital tract in juvenile Eastern Fence Lizards (Sceloporus undulatus) to better understand the mechanisms underlying the ontogeny of sexual differentiation in reptiles. We examined gonads, mesonephric kidneys and ducts (male reproductive tract primordia), paramesonephric ducts (oviduct primordia), sexual segments of the kidneys (SSKs), and hemiphalluses to determine which structures were sexually dimorphic independent of T treatment and which structures exhibited sexually dimorphic responses to T. To better understand tissue-level responsiveness to T treatment, we also characterized androgen receptor (AR) expression by immunohistochemistry. At approximately 4 months after hatching in control animals, gonads were well differentiated but quiescent; paramesonephric ducts had fully degenerated in males; mesonephric kidneys, mesonephric ducts, and SSKs remained sexually undifferentiated; and hemiphalluses could not be everted in either sex. Exogenous T caused enlargement, regionalization, and secretory activity of the mesonephric ducts and SSKs in both sexes; enlargement and regionalization of the oviducts in females; and enlargement of male hemipenes. The most responsive tissues exhibited moderate but diffuse staining for AR in control lizards and intense nuclear staining in T-treated lizards, suggestive of autoregulation of AR. The similarity between sexes in the responsiveness of the mesonephric ducts and SSK to T indicates an absence of sexually dimorphic organizational effects in these structures prior to treatment, which was initiated approximately 2 months after hatching. In contrast, the sex-specific responses in oviducts and hemipenes indicate that significant organization and/or differentiation had taken place prior to treatment.

Stripes and loss of color in ball pythons (Python regius) are associated with variants affecting endothelin signaling.

Color patterns in nonavian reptiles are beautifully diverse, but little is known about the genetics and development of these patterns. Here, we investigated color patterning in pet ball pythons (Python regius), which have been bred to show color phenotypes that differ dramatically from the wildtype form. We report that several color phenotypes in pet animals are associated with putative loss-of-function variants in the gene encoding endothelin receptor EDNRB1: (1) frameshift variants in EDNRB1 are associated with conversion of the normal mottled color pattern to skin that is almost fully white, (2) missense variants affecting conserved sites of the EDNRB1 protein are associated with dorsal, longitudinal stripes, and (3) substitutions at EDNRB1 splice donors are associated with subtle changes in patterning compared to wildtype. We propose that these phenotypes are caused by loss of specialized color cells (chromatophores), with loss ranging from severe (fully white) to moderate (dorsal striping) to mild (subtle changes in patterning). Our study is the first to describe variants affecting endothelin signaling in a nonavian reptile and suggests that reductions in endothelin signaling in ball pythons can produce a variety of color phenotypes, depending on the degree of color cell loss.

Pulmonary bypass shunt is turned off during walking in the American alligator (Alligator mississippiensis)

In non-avian reptiles, the pulmonary bypass (right-to-left) shunt directs blood away from lungs via the left aorta (LAo). At times of elevated oxygen demand, such as exercise or digestion, animals can boost systemic oxygen delivery by either increasing pulmonary bypass shunting to augment cardiac output or reducing the shunt to maximise arterial oxygen content. Previous studies on squamates and turtles showed the shunt is reduced when oxygen demand rises. In contrast, a recent study showed increased shunting during digestion in the American alligator, but whether a similar response occurs during exercise has not been reported to date. We investigated the effects of terrestrial locomotion on blood flow and shunting pattern in female juveniles of the American alligator (n=6; body mass 700-1100g). Perivascular probes were chronically implanted on the major vessels of the outflow tract: the left aorta (LAo), the left pulmonary artery (LPA), the common carotid artery (CCA), and the right aortic arch jointly with the right subclavian artery (RAo+SCA). Shunt fraction was calculated as LAo flow relative to the right ventricular output (= LAo + 2✕LPA). Experiments were performed in a walk-in environmental chamber on animals fasted for a week and equilibrated to 30°C. Upon instrumentation and recovery from surgery, animals were exercised on a level treadmill (0.5 km/h) until exhaustion. Blood flows were recorded continuously before, during and after (10+ min) exercise. Systolic blood flow in the LAo was in the forward direction (shunt on) in alligators at rest, became retrograde (shunt off) immediately upon onset of exercise, and turned anterograde (shunt on again) during recovery. Compared with flow rate values obtained at rest (1.64±0.30 ml/kg/min), average (systolic and diastolic) flow in the LAo decreased significantly during exercise (0.65±0.46 ml/kg/min), and gradually increased with time during recovery (to 1.52±0.28 ml/kg/min in the 10 th min post-exercise). In contrast, blood flow in other instrumented vessels increased during exercise, and gradually decreased during recovery. Shunt fraction was highly variable in resting animals (0.4 – 11.5%), decreased significantly during exercise, remained depressed in the 1 st min post-exercise and increased gradually during recovery to (p<0.05, mixed model ANOVA, post-hoc Dunnett’s test with rest as control). This suggests that alligators turn the pulmonary bypass shunt off during acute bouts of terrestrial exercise. According to the Fick Principle, crocodilians favour increasing arterial oxygen content, by reducing admixture of oxygenated and deoxygenated bloodstreams, over augmenting cardiac output with shunt flow. Thus, despite profound differences in their heart morphology, crocodilian and non-crocodilian “reptiles” exhibit the same response – turning the shunt off – at times of elevated oxygen demand. Whether this is modulated by exercise-induced sympathetic stimulation remains to be tested. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.