Oxygen and temperature affect cell sizes differently among tissues and between sexes of Drosophila melanogaster.

Spatio-temporal gradients in thermal and oxygen conditions trigger evolutionary and developmental responses in ectotherms' body size and cell size, which are commonly interpreted as adaptive. However, the evidence for cell-size responses is fragmentary, as cell size is typically assessed in single tissues. In a laboratory experiment, we raised genotypes of Drosophila melanogaster at all combinations of two temperatures (16 °C or 25 °C) and two oxygen levels (10% or 22%) and measured body size and the sizes of cells in different tissues. For each sex, we measured epidermal cells in a wing and a leg and ommatidial cells of an eye. For males, we also measured epithelial cells of a Malpighian tubule and muscle cells of a flight muscle. On average, females emerged at a larger body size than did males, having larger cells in all tissues. Flies of either sex emerged at a smaller body size when raised under warm or hypoxic conditions. Development at 25 °C resulted in smaller cells in most tissues. Development under hypoxia resulted in smaller cells in some tissues, especially among females. Altogether, our results show thermal and oxygen conditions trigger shifts in adult size, coupled with the systemic orchestration of cell sizes throughout the body of a fly. The nature of these patterns supports a model in which an ectotherm adjusts its life-history traits and cellular composition to prevent severe hypoxia at the cellular level. However, our results revealed some inconsistencies linked to sex, cell type, and environmental parameters, which suggest caution in translating information obtained for single type of cells to the organism as a whole.

Identifying the best strategy for soccer penalty success: A predictive model for optimising behavioural and biomechanical trade-offs.

Success in a soccer penalty can be the difference between winning and losing matches. The outcome is determined by a complex interaction between the shooter and goalkeeper, whose performances are constrained by biomechanical trade-offs. To overcome these performance constraints, each player has a range of available strategies. Shooters can kick at different speeds, affecting accuracy, while goalkeepers can move at various times (leave-times), affecting the time available to move and the probability they move in the correct direction. Previous models of penalty success ignore such trade-offs and how they interact to influence the outcome. Here, we present a model that accounts for shooting inaccuracy to predict the probability of success for all shooting strategies, defined as any combination of: shot speed, position where the shooter aims, shooter footedness, and kicking technique (side-foot or instep). To estimate the probability of success each shooting strategy is matched against all possible goalkeeper leave-times, considering the probability each leave-time is chosen. We test the model against an average goalkeeper and a goalkeeper who tends to move later. Against the average goalkeeper, aiming on the ground toward the centre of the goal is optimal; however, against a late moving goalkeeper, aiming on the ground to the extremities of the goal is effective, with the optimal target in the horizontal dimension dependent on shot speed, kick technique, and footedness. Coaches could use this model to identify their best penalty takers and each players' optimal shooting strategy against either the average goalkeeper or a specific goalkeeper.

The correlated evolution of foraging mode and reproductive effort in lizards.

Life-history theory suggests that the optimal reproductive effort of an organism is affected by factors such as energy acquisition and predation risk. The observation that some organisms actively search for their prey and others ambush them creates the expectation of different energy needs and predation risk associated with each foraging behaviour, the so-called 'foraging-mode paradigm'. Although this paradigm has been around for decades, the empirical evidence consists of conflicting results derived from competing models based on different mechanisms. For instance, models within the foraging-mode paradigm suggest that widely foraging females have evolved low reproductive effort, because a heavy reproductive load decreases their ability to escape from predators. By contrast, a long-standing prediction of evolutionary theory indicates that organisms subject to high extrinsic mortality, should invest more in reproduction. Here, we present the first partial evidence that widely foraging species have evolved greater reproductive effort than have sit-and-wait species, which we attribute to a larger body size and greater mortality among mobile foragers. According to our findings, we propose a theoretical model that could explain the observed pattern in lizards, suggesting ways for evolutionary ecologists to test mechanistic hypotheses at the intraspecific level.

Macroclimatic and maternal effects on the evolution of reproductive traits in lizards.

Much of life-history theory rests on fundamental assumptions about constraints on the acquisition and allocation of energy to growth and reproduction. In general, the allocation of energy to reproduction depends on maternal size, which in turn depends on environmental factors experienced throughout the life of the mother. Here, we used phylogenetic path analyses to evaluate competing hypotheses about the environmental and maternal drivers of reproductive traits in lizards. In doing so, we discovered that precipitation, rather than temperature, has shaped the evolution of the life history. Specifically, environments with greater rainfall have enabled the evolution of larger maternal size. In turn, these larger mothers produce larger clutches of larger offspring. However, annual precipitation has a negative direct effect on offspring size, despite the positive indirect effect mediated by maternal size. Possibly, the evolution of offspring size was driven by the need to conserve water in dry environments, because small organisms are particularly sensitive to water loss. Since we found that body size variation among lizards is related to a combination of climatic factors, mainly precipitation and perhaps primary production, our study challenges previous generalizations (e.g., temperature-size rule and Bergmann's rule) and suggests alternative mechanisms underlying the evolution of body size.

A chromosome-level genome assembly for the eastern fence lizard (Sceloporus undulatus), a reptile model for physiological and evolutionary ecology.

BACKGROUND: High-quality genomic resources facilitate investigations into behavioral ecology, morphological and physiological adaptations, and the evolution of genomic architecture. Lizards in the genus Sceloporus have a long history as important ecological, evolutionary, and physiological models, making them a valuable target for the development of genomic resources. FINDINGS: We present a high-quality chromosome-level reference genome assembly, SceUnd1.0 (using 10X Genomics Chromium, HiC, and Pacific Biosciences data), and tissue/developmental stage transcriptomes for the eastern fence lizard, Sceloporus undulatus. We performed synteny analysis with other snake and lizard assemblies to identify broad patterns of chromosome evolution including the fusion of micro- and macrochromosomes. We also used this new assembly to provide improved reference-based genome assemblies for 34 additional Sceloporus species. Finally, we used RNAseq and whole-genome resequencing data to compare 3 assemblies, each representing an increased level of cost and effort: Supernova Assembly with data from 10X Genomics Chromium, HiRise Assembly that added data from HiC, and PBJelly Assembly that added data from Pacific Biosciences sequencing. We found that the Supernova Assembly contained the full genome and was a suitable reference for RNAseq and single-nucleotide polymorphism calling, but the chromosome-level scaffolds provided by the addition of HiC data allowed synteny and whole-genome association mapping analyses. The subsequent addition of PacBio data doubled the contig N50 but provided negligible gains in scaffold length. CONCLUSIONS: These new genomic resources provide valuable tools for advanced molecular analysis of an organism that has become a model in physiology and evolutionary ecology.

Oxygen supply limits the chronic heat tolerance of locusts during the first instar only.

Although scientists know that overheating kills many organisms, they do not agree on the mechanism. According to one theory, referred to as oxygen- and capacity-limitation of thermal tolerance, overheating occurs when a warming organism's demand for oxygen exceeds its supply, reducing the organism's supply of ATP. This model predicts that an organism's heat tolerance should decrease under hypoxia, yet most terrestrial organisms tolerate the same amount of warming across a wide range of oxygen concentrations. This point is especially true for adult insects, who deliver oxygen through highly efficient respiratory systems. However, oxygen limitation at high temperatures may be more common during immature life stages, which have less developed respiratory systems. To test this hypothesis, we measured the effects of heat and hypoxia on the survival of South American locusts (Schistocerca cancellata) throughout development and during specific instars. We demonstrate that the heat tolerance of locusts depends on oxygen supply during the first instar but not during later instars. This finding provides further support for the idea that oxygen limitation of thermal tolerance depends on respiratory performance, especially during immature life stages.

Oxygen supply limits the heat tolerance of avian embryos.

Physiologists have primarily focused on two potential explanations for heat stress in animals-the classic model of molecular stability and an alternative model of oxygen limitation. Although the classic model has widespread support, the oxygen-supply model applies to many aquatic animals and some terrestrial ones. In particular, the embryonic stage of terrestrial animals seems most susceptible to oxygen limitation because embryos acquire oxygen from the atmosphere by diffusion rather than ventilation. We report experiments confirming the two conditions of the oxygen-supply model in Japanese quail embryos, Coturnix coturnix. Hypoxia (12% O2) greatly reduced the chance of survival at 47.5°C, and hyperoxia greatly improved the chance of survival at 48.5°C. This finding expands the scope of the oxygen-supply model to a terrestrial, endothermic species, suggesting that oxygen supply generally limits the heat tolerance of embryos.

Oxygen Limitation Does Not Drive the Decreasing Heat Tolerance of Grasshoppers during Development.

Thermal physiology changes as organisms grow and develop, but we do not understand what causes these ontogenetic shifts. According to the theory of oxygen- and capacity-limited thermal tolerance, an organism's heat tolerance should change throughout ontogeny as its ability to deliver oxygen varies. As insects grow during an instar, their metabolic demand increases without a proportional increase in the size of tracheae that supply oxygen to the tissues. If oxygen delivery limits heat tolerance, the mismatch between supply and demand should make insects more susceptible to heat and hypoxia as they progress through an instar. We tested this hypothesis by measuring the heat tolerance of grasshoppers (Schistocerca americana) on the second and seventh days of the sixth instar, in either a normoxic or a hypoxic atmosphere (21% or 10% O2, respectively). As expected, heat tolerance decreased as grasshoppers grew larger. Yet contrary to expectation, hypoxia had no effect on heat tolerance across all stages and sizes. Although heat tolerance declines as grasshoppers grow, this pattern must stem from a mechanism other than oxygen limitation.

The World Still Is Not Flat: Lessons Learned from Organismal Interactions with Environmental Heterogeneity in Terrestrial Environments.

Over the past decade, ecologists and physiologists alike have acknowledged the importance of environmental heterogeneity. Meaningful predictions of the responses of organisms to climate will require an explicit understanding of how organismal behavior and physiology are affected by such heterogeneity. Furthermore, the responses of organisms themselves are quite heterogeneous: physiology and behavior vary over different time scales and across different life stages, and because physiological systems do not operate in isolation of one another, they need to be considered in a more integrated fashion. Here, we review case studies from our laboratories to highlight progress that has been made along these fronts and generalizations that might be made to other systems, particularly in the context of predicting responses to climate change.

Fundamental Flaws with the Fundamental Niche.

For more than 70 years, Hutchinson's concept of the fundamental niche has guided ecological research. Hutchinson envisioned the niche as a multidimensional hypervolume relating the fitness of an organism to relevant environmental factors. Here, we challenge the utility of the concept to modern ecologists, based on its inability to account for environmental variation and phenotypic plasticity. We have ample evidence that the frequency, duration, and sequence of abiotic stress influence the survivorship and performance of organisms. Recent work shows that organisms also respond to the spatial configuration of abiotic conditions. Spatiotemporal variation of the environment interacts with the genotype to generate a unique phenotype at each life stage. These dynamics cannot be captured adequately by a multidimensional hypervolume. Therefore, we recommend that ecologists abandon the niche as a tool for predicting the persistence of species and embrace mechanistic models of population growth that incorporate spatiotemporal dynamics.

Thermal acclimation of flies from three populations of Drosophila melanogaster fails to support the seasonality hypothesis.

In seasonal environments, natural selection should favor genotypes that acclimate to slow and predictable changes in temperature. Selective pressure for acclimation should be especially strong for animals that complete many generations per year, because seasonal warming or cooling causes offspring to experience different temperatures than their parents did. Here, we studied variation in acclimation capacity among three populations of Drosophila melanogaster. We used a reverse acclimation design to see whether developmental acclimation persisted throughout adulthood. Flies developed from fertilization to adulthood at either 16° or 26 °C. Then, flies either remained at the same temperature or moved to the other temperature for 7 days. We measured fecundity at seven temperatures ranging from 14° to 36°C. Genotypes from North Carolina and Vermont laid more eggs at 16 °C after spending the larval and adult stages at 16 °C, instead of 26 °C. In both populations, the benefit of acclimation to 16 °C during development was erased by acclimation to 26 °C during adulthood. In contrast to our prediction, genotypes from Indiana laid fewer eggs at 16 °C or 26 °C after developing at this temperature. Overall, these data provide only weak support for the models of optimal acclimation in seasonal environments.

The origin and maintenance of metabolic allometry in animals.

Organisms vary widely in size, from microbes weighing 0.1 pg to trees weighing thousands of megagrams - a 1021-fold range similar to the difference in mass between an elephant and the Earth. Mass has a pervasive influence on biological processes, but the effect is usually non-proportional; for example, a tenfold increase in mass is typically accompanied by just a four- to sevenfold increase in metabolic rate. Understanding the cause of allometric scaling has been a long-standing problem in biology. Here, we examine the evolution of metabolic allometry in animals by linking microevolutionary processes to macroevolutionary patterns. We show that the genetic correlation between mass and metabolic rate is strong and positive in insects, birds and mammals. We then use these data to simulate the macroevolution of mass and metabolic rate, and show that the interspecific relationship between these traits in animals is consistent with evolution under persistent multivariate selection on mass and metabolic rate over long periods of time.

The neuroscience of adaptive thermoregulation.

The nervous system acts as a biological thermostat by controlling behaviors that regulate the warming and cooling of animals. We review the structures responsible for thermoregulation in three model species: roundworms (Caenorhabditis elegans), flies (Drosophila melanogaster), and rats (Rattus novegicus). We then consider additional features of the nervous system required to explain adaptive plasticity of the set-point temperature and the precision of thermoregulation. Because animals use resources such as energy, water, and oxygen to thermoregulate, the nervous system monitors the abundance of these resources and adjusts the strategy of thermoregulation accordingly. Starvation, dehydration, or hypoxemia alter the activity of temperature-sensitive neurons in the pre-optic area of the hypothalamus. Other regions of the brain work in conjunction with the hypothalamus to promote adaptive plasticity of thermoregulation. For example, the amygdala likely inhibits neurons of the pre-optic area, overriding thermoregulation when a risk of predation or a threat of aggression exists. Moreover, the hippocampus enables an animal to remember microhabitats that enable safe and effective thermoregulation. In ectothermic animals, such as C. elegans and D. melanogaster, the nervous system can alter set-point temperatures as the environmental temperatures change. To build on this knowledge, neuroscientists can use experimental evolution to study adaptation of neural phenotypes in controlled thermal environments. A microevolutionary perspective would leverage our understanding of ecological processes to predict the origin and maintenance of neural phenotypes by natural selection.

Lizards perceived abiotic and biotic stressors independently when competing for shade in terrestrial mesocosms.

Hormones such as glucocorticoids and androgens enable animals to respond adaptively to environmental stressors. For this reason, circulating glucocorticoids became a popular biomarker for estimating the quality of an environment, and circulating androgens are frequently used to indicate social dominance. Here, we show that access to thermal resources influence the hormones and behavior of male lizards (Sceloporus jarrovi). We exposed isolated and paired males to different thermal landscapes, ranging from one large patch of shade to sixteen smaller patches. Both the presence of a competitor and the patchiness of the thermal environment influenced hormone concentrations and movement patterns. When shade was concentrated in space, paired lizards competed more aggressively and circulated more corticosterone. Even without competitors, lizards circulated more corticosterone in landscapes with fewer patches of shade. Conversely, shifts in circulating testosterone depended only on the relative body size of a lizard; when paired, large males and small males circulated more and less testosterone, respectively. Furthermore, isolated males moved the farthest and covered the most area when shade was concentrated in a single patch, but paired males did the opposite. Because the total area of shade in each landscape was the same, these hormonal and behavioral responses of lizards reflect the ability to access shade. Thus, circulating glucocorticoids should reflect the thermal quality of an environment when researchers have controlled for other factors. Moreover, a theory of stress during thermoregulation would help ecologists anticipate physiological and behavioral responses to changing climates.

Behaviors of shooter and goalkeeper interact to determine the outcome of soccer penalties.

During a soccer penalty, the shooter's strategy and the goalkeeper's strategy interact to determine the outcome. However, most models of penalty success overlook its interactive nature. Here, we quantified aspects of shooter and goalkeeper strategies that interact to influence the outcome of soccer penalties-namely, how the speed of the shot affects the goalkeeper's leave time or shot-blocking success, and the effectiveness of deceptive strategies. We competed 7 goalkeepers and 17 shooters in a series of penalty shoot-out competitions with a total of 1278 shots taken. Each player was free to use any strategy within the rules of a penalty shot, and game-like pressure was created via monetary incentive for goal-scoring (or blocking). We found that faster shots lead to earlier leave times and were less likely blocked by goalkeepers, and-unlike most previous studies-that deceptive shooting strategies did not decrease the likelihood goalkeepers moved in the correct direction. To help identify optimal strategies for shooters and goalkeepers, we generated distributions and mathematical functions sport scientists can use to develop more comprehensive models of penalty success.

Anticipating the Direction of Soccer Penalty Shots Depends on the Speed and Technique of the Kick.

To succeed at a sport, athletes must manage the biomechanical trade-offs that constrain their performance. Here, we investigate a previously unknown trade-off in soccer: how the speed of a kick makes the outcome more predictable to an opponent. For this analysis, we focused on penalty kicks to build on previous models of factors that influence scoring. More than 700 participants completed an online survey, watching videos of penalty shots from the perspective of a goalkeeper. Participants (ranging in soccer playing experience from never played to professional) watched 60 penalty kicks, each of which was occluded at a particular moment (-0.4 s to 0.0 s) before the kicker contacted the ball. For each kick, participants had to predict shot direction toward the goal (left or right). As expected, predictions became more accurate as time of occlusion approached ball contact. However, the effect of occlusion was more pronounced when players kicked with the side of the foot than when they kicked with the top of the foot (instep). For side-foot kicks, the direction of shots was predicted more accurately for faster kicks, especially when a large portion of the kicker's approach was presented. Given the trade-off between kicking speed and directional predictability, a penalty kicker might benefit from kicking below their maximal speed.

Measuring behavioral thermal tolerance to address hot topics in ecology, evolution, and conservation.

Understanding the impacts of anthropogenic climate change requires knowing how animals avoid heat stress, and the consequences of failing to do so. Animals primarily use behavior to avoid overheating, but biologists' means for measuring and interpreting behavioral signs of stress require more development. Herein, we develop the measurement of behavioral thermal tolerance using four species of lizards. First, we adapt the voluntary thermal maximum concept (VTM) to facilitate its measurement, interpretation, and comparison across species. Second, we evaluate the sensitivity of the VTM to diverse measurement options (warming rate, time of day, etc) across four species with highly different life histories. Finally, we clarify the interpretation of VTM in two ways. First, we show the effects of exposure to the VTM on panting behavior, mass loss, and locomotor function loss of two species. Second, we compared the VTM with the preferred body temperatures (PBT) and critical thermal maximum (CTMAX) intraspecifically. We found that the VTM can be consistently estimated through different methods and methodological options, only very slow warming rates affected its estimates in one species. Exposure to the VTM caused panting between 5 and 50 min and induced exceptionally high mass loss rates. Loss of locomotion function started after 205 min. Further, the VTM did not show intraspecific correlations with the PBT and CTMAX. Our study suggests the VTM is a robust and flexible measure of thermal tolerance and highlights the need for multispecies evaluations of thermal indices. The lack of correlation between the VTM, the PBT and CTMAX suggests the VTM may evolve relatively free between the other parameters. We make reccommendations for understanding and using the VTM in studies of ecology, evolution, and conservation.

Modeling the two-dimensional accuracy of soccer kicks.

In many sports, athletes perform motor tasks that simultaneously require both speed and accuracy for success, such as kicking a ball. Because of the biomechanical trade-off between speed and accuracy, athletes must balance these competing demands. Modelling the optimal compromise between speed and accuracy requires one to quantifyhow task speed affects the dispersion around a target, a level of experimental detail not previously addressed. Using soccer penalties as a system, we measured two-dimensional kicking error over a range of speeds, target heights, and kicking techniques. Twenty experienced soccer players executed a total of 8466 kicks at two targets (high and low). Players kicked with the side of their foot or the instep at ball speeds ranging from 40% to 100% of their maximum. The inaccuracy of kicks was measured in horizontal and vertical dimensions. For both horizontal and vertical inaccuracy, variance increased as a power function of speed, whose parameter values depended on the combination of kicking technique and target height. Kicking precision was greater when aiming at a low target compared to a high target. Side-foot kicks were more accurate than instep kicks. The centre of the dispersion of shots shifted as a function of speed. An analysis of the covariance between horizontal and vertical error revealed right-footed kickers tended to miss below and to the left of the target or above and to the right, while left-footed kickers tended along the reflected axis. Our analysis provides relationships needed to model the optimal strategy for penalty kickers.

Oxygen supply did not affect how lizards responded to thermal stress.

Zoologists rely on mechanistic niche models of behavioral thermoregulation to understand how animals respond to climate change. These models predict that species will need to disperse to higher altitudes to persist in a warmer world. However, thermal stress and, thus, thermoregulatory behavior may depend on atmospheric oxygen as well as environmental temperatures. Severe hypoxia causes animals to prefer lower body temperatures, which could be interpreted as evidence that oxygen supply limits heat tolerance. Such a constraint could prevent animals from successfully dispersing to high elevations during climate change. Still, an effect of oxygen supply on preferred body temperature has only been observed when oxygen concentrations fall far below levels experienced in nature. To see whether animals perceive greater thermal stress at an ecologically relevant level of hypoxia, we studied the thermoregulatory behavior of lizards (Sceloporus tristichus) exposed to oxygen concentrations of 13% and 21% (equivalent to PO2 at 4000 m and 0 m, respectively). In addition, we exposed lizards to 29% oxygen to see whether they would accept a higher body temperature at hyperoxia than at normoxia. At each oxygen level, we measured a behavioral response to heat stress known as the voluntary thermal maximum: the temperature at which a warming animal sought a cool refuge. Oxygen concentration had no discernable effect on the voluntary thermal maximum, suggesting that lizards experience thermal stress similarly at all 3 levels of oxygen (13%, 12% and 29%). Future research should focus on thermoregulatory behaviors under ecologically relevant levels of hypoxia.

Skill not athleticism predicts individual variation in match performance of soccer players.

Just as evolutionary biologists endeavour to link phenotypes to fitness, sport scientists try to identify traits that determine athlete success. Both disciplines would benefit from collaboration, and to illustrate this, we used an analytical approach common to evolutionary biology to isolate the phenotypes that promote success in soccer, a complex activity of humans played in nearly every modern society. Using path analysis, we quantified the relationships among morphology, balance, skill, athleticism and performance of soccer players. We focused on performance in two complex motor activities: a simple game of soccer tennis (1 on 1), and a standard soccer match (11 on 11). In both contests, players with greater skill and balance were more likely to perform better. However, maximal athletic ability was not associated with success in a game. A social network analysis revealed that skill also predicted movement. The relationships between phenotypes and success during individual and team sports have potential implications for how selection acts on these phenotypes, in humans and other species, and thus should ultimately interest evolutionary biologists. Hence, we propose a field of evolutionary sports science that lies at the nexus of evolutionary biology and sports science. This would allow biologists to take advantage of the staggering quantity of data on performance in sporting events to answer evolutionary questions that are more difficult to answer for other species. In return, sports scientists could benefit from the theoretical framework developed to study natural selection in non-human species.