Sex-specific effects of immune challenges on green anole lizard metabolism.

Immune responses can increase survival, but they can also incur a variety of costs that may lead to phenotypic trade-offs. The nature of trade-offs between immune activity and other components of the phenotype can vary and depend on the type and magnitude of immune challenge, as well as the energetic costs of simultaneously expressing other traits. There may also be sex-specific differences in both immune activity and trade-offs, particularly with regard to energy expenditure that might differ between males and females during the breeding season. Females are generally expected to invest less in nonspecific immune responses compared to males due to differences in the allocation of resources to reproduction, which may lead to sex differences in the metabolic costs of immunity. We tested for sex-specific differences in metabolic costs of different types of immune challenges in Anolis carolinensis lizards, including lipopolysaccharide (LPS) injection and wounding. We also tested for differences in immune prioritization by measuring bacterial killing ability (BKA). We predicted males would show a greater increase in metabolism after immune challenges, with combined immune challenges eliciting the greatest response. Furthermore, we predicted that metabolic costs would result in decreased BKA. LPS injection increased the resting metabolic rate (RMR) of males but not females. Wounding did not affect RMR of either sex. However, there was an inverse relationship between BKA and wound healing in LPS-injected lizards, suggesting dynamic tradeoffs among metabolism and components of the immune system.

How does mitochondria function contribute to aerobic performance enhancement in lizards?

Aims: Aerobic exercise typically enhances endurance across vertebrates so that chronically high energy demands can be met. Some known mechanisms of doing this include increases in red blood cell numbers, angiogenesis, muscle fiber adaptions, mitochondria biogenesis, and changes to cellular metabolism and oxidative phosphorylation. We used green anole lizards (Anolis carolinensis) to test for an effect of aerobic exercise on metabolism, mitochondria densities, and mitochondrial function. Methods: We first tested the response of green anoles to endurance training and pyrroloquinoline quinone (PQQ) supplementation, which has been shown to increase mitochondria biogenesis. We also conducted a mitochondrial stress test to determine how training affected mitochondrial function in skeletal muscle fibers. Results: Aerobic exercise led to increased endurance and decreased standard metabolic rate (SMR), while PQQ did not affect endurance and increased SMR. In a second experiment, aerobic exercise increased endurance and decreased resting metabolic rate (RMR) in both male and female green anoles. Higher counts of mitochondrial gene copies in trained lizards suggested additional mitochondria adaptations to achieve increased endurance and decreased metabolism. A mitochondrial stress test revealed no effect on baseline oxygen consumption rates of muscle fibers, but untrained lizards had higher maximal oxygen consumption rates with the addition of metabolic fuel. Conclusion: It is likely that trained lizards exhibited lower maximal oxygen consumption rates by developing higher mitochondria efficiency. This adaptation allows for high ATP demand to be met by making more ATP per oxygen molecule consumed. On the other hand, it is possible that untrained lizards prioritized limiting reactive oxygen species (ROS) production at rest, while sacrificing higher levels of proton leak and higher oxygen consumption rates when working to meet high ATP demand.

Sprint training interacts with body mass to affect hepatic insulin-like growth factor expression in female green anoles (Anolis carolinensis).

Locomotor performance is a key predictor of fitness in many animal species. As such, locomotion integrates the output of a number of morphological, physiological, and molecular levels of organization, yet relatively little is known regarding the major molecular pathways that bolster locomotor performance. One potentially relevant pathway is the insulin and insulin-like signaling (IIS) network, a significant regulator of physiological processes such as reproduction, growth, and metabolism. Two primary hormones of this network, insulin-like growth factor 1 (IGF1) and insulin-like growth factor 2 (IGF2) are important mediators of these processes and, consequently, of life-history strategies. We sprint-trained green anole (Anolis carolinensis) females to test the responsiveness of IGF1 and IGF2 hepatic gene expression to exercise training. We also tested how sprint training would affect glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and eukaryotic elongation factor 2 (EEF2). The former is a crucial enzyme for glycolytic function in a cell, and the latter is necessary for protein synthesis. Resistance exercise forces animals to increase investment of resources towards skeletal muscle growth. Because IGF1 and IGF2 are important hormones for growth, and GAPDH and EEF2 are crucial for proper cellular function, we hypothesized that these four genes would be affected by sprint training. We found that sprint training affects IGF and EEF2 expression, such that larger sprint-trained lizards express hepatic IGF1, IGF2, and EEF2 to a lesser extent than similarly sized untrained lizards. These results demonstrate that the IIS, and pathways connected to it, can react in a size-dependent manner and are implicated in the exercise response in reptiles.

Conserved and convergent mechanisms underlying performance-life-history trade-offs.

Phenotypic trade-offs are inevitable in nature, but the mechanisms driving them are poorly understood. Movement and oxygen are essential to all animals, and as such, the common ancestor to all living animals passed on mechanisms to acquire oxygen and contract muscle, sometimes at the expense of other activities or expression of traits. Nevertheless, convergent pathways have also evolved to deal with critical trade-offs that are necessary to survive ubiquitous environmental challenges. We discuss how whole-animal performance traits, such as locomotion, are important to fitness, yet costly, resulting in trade-offs with other aspects of the phenotype via specific conserved and convergent mechanistic pathways across all animals. Specifically, we discuss conserved pathways involved in muscle structure and signaling, insulin/insulin-like signaling, sirtuins, mitochondria and hypoxia-inducible factors, as well as convergent pathways involved in energy regulation, development, reproductive investment and energy storage. The details of these mechanisms are only known from a few model systems, and more comparative studies are needed. We make two main recommendations as a framework for future studies of animal form and function. First, studies of performance should consider the broader life-history context of the organism, and vice versa, as performance expression can require a large portion of acquired resources. Second, studies of life histories or mechanistic pathways that measure performance should do so in meaningful and standardized ways. Understanding proximate mechanisms of phenotypic trade-offs will not only better explain the phenotypes of the organisms we study, but also allow predictions about phenotypic variation at the evolutionary scale.

Immune activation affects whole-organism performance in male but not female green anole lizards (Anolis carolinensis).

Immune responses are intuitively beneficial, but they can incur a variety of costs, many of which are poorly understood. The nature and extent of trade-offs between immune activity and other components of the integrated phenotype can vary, and depend on the type of immune challenge, as well as the energetic costs of simultaneously expressing other traits. There may also be sex differences in both immune activity and immunity-induced trade-offs, particularly in the case of trade-offs involving functional traits such as whole-organism performance capacities that might be of different fitness value to males and females. We tested the response of three performance traits (sprinting, endurance, and biting) to two different immune challenges (LPS injection and wound healing) in both male and female Anolis carolinensis lizards. We found clear differences in how male and female performance capacities were affected by immune activation. LPS injection and wound healing had interactive effects on all three performance traits in males, but immune activation did not affect female performance. We also found that the degree of wound healing exhibited complex interactive effects involving sex and type of immune activation that varied depending on the performance trait in question. These results demonstrate that male and female green anoles experience different consequences of immune responses, and also that the type and extent of that activation can drive trait-specific performance trade-offs.

Life history and environment predict variation in testosterone across vertebrates.

Endocrine systems act as key intermediaries between organisms and their environments. This interaction leads to high variability in hormone levels, but we know little about the ecological factors that influence this variation within and across major vertebrate groups. We study this topic by assessing how various social and environmental dynamics influence testosterone levels across the entire vertebrate tree of life. Our analyses show that breeding season length and mating system are the strongest predictors of average testosterone concentrations, whereas breeding season length, environmental temperature, and variability in precipitation are the strongest predictors of within-population variation in testosterone. Principles from small-scale comparative studies that stress the importance of mating opportunity and competition on the evolution of species differences in testosterone levels, therefore, likely apply to the entire vertebrate lineage. Meanwhile, climatic factors associated with rainfall and ambient temperature appear to influence variability in plasma testosterone, within a given species. These results, therefore, reveal how unique suites of ecological factors differentially explain scales of variation in circulating testosterone across mammals, birds, reptiles, amphibians, and fishes.

Endurance and sprint training affect immune function differently in green anole lizards (Anolis carolinensis).

Limited resources must be partitioned among traits that enhance fitness. Although survival-related traits often trade off with reproduction, survival-related traits themselves may trade off with each other under energy limitations. Whole-organism performance and the immune system both enhance survival, yet are costly, but it is unclear how the two might trade off with each other under energy-limited conditions. Resources can be allocated to very different types of performance (e.g. aerobic endurance versus anaerobic sprinting), just as they can be allocated to different components of the immune system (e.g. innate versus acquired) to maximize survival. We forced allocation to different performance traits in green anole lizards (Anolis carolinensis) using specialized exercise training, to determine how different components of the immune system would be impacted by shifts in energy use. We measured immunocompetence in endurance-trained, sprint-trained and untrained control lizards by evaluating swelling response to phytohemagglutinin (cell-mediated immunity), antibody response to sheep red blood cells (acquired humoral immunity) and wound healing (integrated immunity). Endurance-trained lizards had reduced cell-mediated immunity, whereas sprint-trained lizards had reduced rates of wound healing. The acquired immune response was not affected by either type of training. Because each immune measure responded differently to the different types of training, our results do not support the hypothesis that simple energy limitation determines overall investment in immunity. Instead, different components of the immune system appear to be affected in ways specific to how energy is invested in performance.

Baseline and stress-induced corticosterone levels across birds and reptiles do not reflect urbanization levels.

Rates of human-induced environmental change continue increasing with human population size, potentially altering animal physiology and negatively affecting wildlife. Researchers often use glucocorticoid concentrations (hormones that can be associated with stressors) to gauge the impact of anthropogenic factors (e.g. urbanization, noise and light pollution). Yet, no general relationships between human-induced environmental change and glucocorticoids have emerged. Given the number of recent studies reporting baseline and stress-induced corticosterone (the primary glucocorticoid in birds and reptiles) concentrations worldwide, it is now possible to conduct large-scale comparative analyses to test for general associations between disturbance and baseline and stress-induced corticosterone across species. Additionally, we can control for factors that may influence context, such as life history stage, environmental conditions and urban adaptability of a species. Here, we take a phylogenetically informed approach and use data from HormoneBase to test if baseline and stress-induced corticosterone are valid indicators of exposure to human footprint index, human population density, anthropogenic noise and artificial light at night in birds and reptiles. Our results show a negative relationship between anthropogenic noise and baseline corticosterone for birds characterized as urban avoiders. While our results potentially indicate that urban avoiders are more sensitive to noise than other species, overall our study suggests that the relationship between human-induced environmental change and corticosterone varies across species and contexts; we found no general relationship between human impacts and baseline and stress-induced corticosterone in birds, nor baseline corticosterone in reptiles. Therefore, it should not be assumed that high or low levels of exposure to human-induced environmental change are associated with high or low corticosterone levels, respectively, or that closely related species, or even individuals, will respond similarly. Moving forward, measuring alternative physiological traits alongside reproductive success, health and survival may provide context to better understand the potential negative effects of human-induced environmental change.

Sprint speed is unaffected by dietary manipulation in trained male Anolis carolinensis lizards.

Performance traits are energetically costly, and their expression and use can drive trade-offs with other energetically costly life-history traits. However, different performance traits incur distinct costs and may be sensitive to both resource limitation and to the types of resources that are accrued. Protein is likely to be especially important for supporting burst performance traits such as sprint speed, but the effect of varying diet composition on sprint training in lizards, an emerging model system for exercise training, is unknown. We tested the hypothesis that the response to sprint training is sensitive to both the type and amount of resources in Anolis carolinensis. We also measured bite force across all treatments as a control whole-organism performance trait that should be unaffected by locomotor training. Both mass and bite force are reduced by dietary restriction over the course of 9 weeks of sprint training, but sprint speed is unaffected by either training or dietary restriction relative to controls. Furthermore, protein supplementation does not rescue a decline in either mass or bite force in trained, diet-restricted males. These results contrast with those for endurance training, and suggest that sprint speed is more canalized than either endurance or bite force in green anoles.

Corticosterone in Lizard Egg Yolk Is Reduced by Maternal Diet Restriction but Unaltered by Maternal Exercise.

When females face adverse environmental conditions, physiological changes, such as elevated corticosterone levels, to cope with the stressors may also impact their offspring. Such maternal effects are often considered adaptive and may "prime" the offspring for the same adverse environment, but maternal corticosterone levels do not always match that of the eggs produced. We examined how diet restriction and increased locomotor activity, via exercise training, affected steroid hormone levels of female green anole lizards, as well as the hormone levels in the yolk of their eggs. Diet restriction did not affect female hormone levels, but training increased corticosterone levels. Despite this, training did not affect yolk steroid levels, but eggs from females with diet restriction had lower corticosterone levels in yolk. This suggests that two common stressors, food shortage and increased locomotor activity, impact female physiology in a way that is not translated to her offspring.

Macroevolutionary Patterning in Glucocorticoids Suggests Different Selective Pressures Shape Baseline and Stress-Induced Levels.

Glucocorticoid (GC) hormones are important phenotypic mediators across vertebrates, but their circulating concentrations can vary markedly. Here we investigate macroevolutionary patterning in GC levels across tetrapods by testing seven specific hypotheses about GC variation and evaluating whether the supported hypotheses reveal consistent patterns in GC evolution. If selection generally favors the "supportive" role of GCs in responding effectively to challenges, then baseline and/or stress-induced GCs may be higher in challenging contexts. Alternatively, if selection generally favors "protection" from GC-induced costs, GCs may be lower in environments where challenges are more common or severe. The predictors of baseline GCs were all consistent with supportive effects: levels were higher in smaller organisms and in those inhabiting more energetically demanding environments. During breeding, baseline GCs were also higher in populations and species with fewer lifetime opportunities to reproduce. The predictors of stress-induced GCs were instead more consistent with the protection hypothesis: during breeding, levels were lower in organisms with fewer lifetime reproductive opportunities. Overall, these patterns indicate a surprising degree of consistency in how some selective pressures shape GCs across broad taxonomic scales; at the same time, in challenging environments selection appears to operate on baseline and stress-induced GCs in distinct ways.

Experimentally enhanced performance decreases survival in nature.

Superior locomotor performance confers advantages in terms of male combat success, survival and fitness in a variety of organisms. In humans, investment in increased performance via the exercise response is also associated with numerous health benefits, and aerobic capacity is an important predictor of longevity. Although the response to exercise is conserved across vertebrates, no studies have tested whether non-human animals that invest in increased athletic performance through exercise realize a survival advantage in nature. Green anole lizards respond to exercise training, and enhanced performance drives trade-offs with reproduction and immunocompetence. We released sprint-trained, endurance-trained and untrained-control male and female green anole lizards into an isolated, urban island in New Orleans, LA, USA and monitored their survival. Sedentary controls realized a significant survivorship advantage compared to trained lizards. Our results suggest that locomotor capacity is currently optimized to maximize survival in green anoles, and that forcing additional investment in performance moves them into a suboptimal phenotypic space relative to their current environmental demands.

Leptin ameliorates the immunity, but not reproduction, trade-off with endurance in lizards.

Life-history trade-offs result from allocation of limited energetic resources to particular traits at the expense of others. When resources are scarce, some traits will take priority over others in the degree of their expression. For example, the current reproduction may be sacrificed to enhance survival. Although intuitive from an evolutionary perspective, such priorities must be based on proximate mechanisms that respond to the current conditions. The hormone leptin serves as a signal of energy availability in vertebrates, and has been proposed as a mediator of energy allocation between reproduction and traits that enhance survival, such as the immune system. However, since leptin affects reproduction and immunity in a similar way, it remains unclear which takes priority when energy availability is low. Green anole lizards (Anolis carolinensis) with increased activity, via exercise training, have a marked decrease in immune function as well as reproduction, especially when calories are restricted. We hypothesized that endurance training and calorie restriction would lower immune and reproductive function due to energy limitation, and supplemental leptin would 'rescue' either immune function or reproduction (or both) due to the hormonal signal that energetic resources are available. We found that supplementary leptin rescued immune function in calorie-restricted, trained lizards, but reproduction was not rescued in males or females. This suggests that immune function and reproduction have different sensitivities to leptin in both sexes, or that reproduction is more energy limited and takes low priority even when the signal of energy availability is present.

Illuminating Endocrine Evolution: The Power and Potential of Large-Scale Comparative Analyses.

Hormones are central mediators of genotype-phenotype and organism-environment interactions. Despite these important functions, the role of selection in shaping hormonal mediators of phenotype remains poorly understood. Thanks to decades of work by endocrinologists, circulating hormone levels have been measured in a diversity of organisms. Variation in other endocrine traits and mediators (e.g., receptor expression and binding globulins), and the hormonal response to standardized challenges (e.g., restraint, pharmacological challenges) are also increasingly measured in both captive and free-living populations. Large-scale comparative analyses of the multitude of available endocrine data represent a particularly promising approach to addressing the function and evolution of these key phenotypic mediators, and their potential to serve as indicators of disturbance. Several recent phylogenetic comparative analyses and meta-analyses have begun to reveal the power and potential of these approaches to address key questions in integrative biology. Here we highlight two recent developments that are facilitating such analyses: increasingly powerful and flexible phylogenetic comparative methods, and the release of two endocrine trait databases-HormoneBase (currently 474 species) and the Wildlife Endocrinology Information Network (currently 25 species)-that contain compiled measures of endocrine traits across vertebrates. Increasingly comprehensive comparative analyses of endocrine data could provide insight into many interesting questions, including how rapidly changing environments are impacting phenotypes, why endocrine traits differ so remarkably within and across populations, and the evolution of plasticity. The endocrine system mediates interactions between genotypes and phenotypes, and between organisms and their environment. Environmentally induced hormonal responses regulate phenotypic flexibility across timescales by altering physiological state, gene expression, and epigenetic marks. A staggering diversity of phenotypic traits are mediated by hormones from early development through senescence. Through their actions on behavior, hormones also exert widespread influence over how organisms interact with their biotic and abiotic environments. Because hormones are responsive to the environment, there has long been interest in their use as biomarkers of exposure to challenges. More recently, increasing attention has been paid to the potential for within and among-population variation in endocrine regulation or responsiveness to serve as indicators of resistance or resilience to future challenges, or measures of evolutionary potential.

Species-Specific Means and Within-Species Variance in Glucocorticoid Hormones and Speciation Rates in Birds.

At macroevolutionary scales, stress physiology may have consequences for species diversification and subspecies richness. Populations that exploit new resources or undergo range expansion should cope with new environmental challenges, which could favor higher mean stress responses. Within-species variation in the stress response may also play a role in mediating the speciation process: in species with broad variation, there will always be some individuals that can tolerate an unpredictable environment, whereas in species with narrow variation there will be fewer individuals that are able to thrive in a new ecological niche. We tested for the evolutionary relationship between stress response, speciation rate, and subspecies richness in birds by relying on the HormoneBase repository, from which we calculated within- and among-species variation in baseline (BL) and stress-induced (SI) corticosterone levels. To estimate speciation rates, we applied Bayesian analysis of macroevolutionary mixtures that can account for variation in diversification rate among clades and through time. Contrary to our predictions, lineages with higher diversification rates were not characterized by higher BL or SI levels of corticosterone either at the tips or at the deeper nodes of the phylogeny. We also found no association between mean hormone levels and subspecies richness. Within-species variance in corticosterone levels showed close to zero repeatability, thus it is highly unlikely that this is a species-specific trait that influences diversification rates. These results imply that stress physiology may play a minor, if any, role in determining speciation rates in birds.

Standing Variation and the Capacity for Change: Are Endocrine Phenotypes More Variable Than Other Traits?

Circulating steroid hormone levels exhibit high variation both within and between individuals, leading some to hypothesize that these phenotypes are more variable than other morphological, physiological, and behavioral traits. This should have profound implications for the evolution of steroid signaling systems, but few studies have examined how endocrine variation compares to that of other traits or differs among populations. Here we provide such an analysis by first exploring how variation in three measures of corticosterone (CORT)-baseline, stress-induced, and post-dexamethasone injection-compares to variation in key traits characterizing morphology (wing length, mass), physiology (reactive oxygen metabolite concentration [d-ROMs] and antioxidant capacity), and behavior (provisioning rate) in two populations of tree swallow (Tachycineta bicolor). After controlling for measurement precision and within-individual variation, we found that only post-dex CORT was more variable than all other traits. Both baseline and stress-induced CORT exhibit higher variation than antioxidant capacity and provisioning rate, but not oxidative metabolite levels or wing length. Variation in post-dex CORT and d-ROMs was also elevated in the higher-latitude population in that inhabits a less predictable environment. We next studied how these patterns might play out on a macroevolutionary scale, assessing patterns of variation in baseline testosterone (T) and multiple non-endocrine traits (body length, mass, social display rate, and locomotion rate) across 17 species of Anolis lizards. At the macroevolutionary level, we found that circulating T levels and the rate of social display output are higher than other behavioral and morphological traits. Altogether, our results support the idea that within-population variability in steroid levels is substantial, but not exceptionally higher than many other traits that define animal phenotypes. As such, circulating steroid levels in free-living animals should be considered traits that exhibit similar levels of variability from individual to individual in a population.

Do Seasonal Glucocorticoid Changes Depend on Reproductive Investment? A Comparative Approach in Birds.

Animals go through different life history stages such as reproduction, moult, or migration, of which some are more energy-demanding than others. Baseline concentrations of glucocorticoid hormones increase during moderate, predictable challenges and thus are expected to be higher when seasonal energy demands increase, such as during reproduction. By contrast, stress-induced glucocorticoids prioritize a survival mode that includes reproductive inhibition. Thus, many species down-regulate stress-induced glucocorticoid concentrations during the breeding season. Interspecific variation in glucocorticoid levels during reproduction has been successfully mapped onto reproductive investment, with species investing strongly in current reproduction (fast pace of life) showing higher baseline and lower stress-induced glucocorticoid concentrations than species that prioritize future reproduction over current attempts (slow pace of life). Here we test the "glucocorticoid seasonal plasticity hypothesis", in which we propose that interspecific variation in seasonal changes in glucocorticoid concentrations from the non-breeding to the breeding season will be related to the degree of reproductive investment (and thus pace of life). We extracted population means for baseline (for 54 species) and stress-induced glucocorticoids (for 32 species) for the breeding and the non-breeding seasons from the database "HormoneBase", also calculating seasonal glucocorticoid changes. We focused on birds because this group offered the largest sample size. Using phylogenetic comparative methods, we first showed that species differed consistently in both average glucocorticoid concentrations and their changes between the two seasons, while controlling for sex, latitude, and hemisphere. Second, as predicted seasonal changes in baseline glucocorticoids were explained by clutch size (our proxy for reproductive investment), with species laying larger clutches showing a greater increase during the breeding season-especially in passerine species. In contrast, changes in seasonal stress-induced levels were not explained by clutch size, but sample sizes were more limited. Our findings highlight that seasonal changes in baseline glucocorticoids are associated with a species' reproductive investment, representing an overlooked physiological trait that may underlie the pace of life.

Metabolic Scaling of Stress Hormones in Vertebrates.

Glucocorticoids (GCs) are stress hormones that can strongly influence physiology, behavior, and an organism's ability to cope with environmental change. Despite their importance, and the wealth of studies that have sought to understand how and why GC concentrations vary within species, we do not have a clear understanding of how circulating GC levels vary within and across the major vertebrate clades. New research has proposed that much interspecific variation in GC concentrations can be explained by variation in metabolism and body mass. Specifically, GC concentrations should vary proportionally with mass-specific metabolic rates and, given known scaling relationships between body mass and metabolic rate, GC concentrations should scale to the -1/4 power of body mass and to the power of 1 with mass-specific metabolic rate. Here, we use HormoneBase, the newly compiled database that includes plasma GC concentrations from free-living and unmanipulated vertebrates, to evaluate this hypothesis. Specifically, we explored the relationships between body mass or mass-specific metabolic rate and either baseline or stress-induced GC (cortisol or corticosterone) concentrations in tetrapods. Our phylogenetically-informed models suggest that, whereas the relationship between GC concentrations and body mass across tetrapods and among mammals is close to -1/4 power, this relationship does not exist in amphibians, reptiles, and birds. Moreover, with the exception of a positive association between stress-induced GC concentrations and mass-specific metabolic rate in birds, we found little evidence that GC concentrations are linked to metabolic rate, although the number of species sampled was quite limited for amphibians and somewhat so for reptiles and mammals. Nevertheless, these results stand in contrast to the generally accepted association between the two and suggest that our observed positive association between body mass and GC concentrations may not be due to the well-established link between mass and metabolism. Large-scale comparative approaches can come with drawbacks, such as pooling and pairing observations from separate sources. However, these broad analyses provide an important counterbalance to the majority of studies examining variation in GC concentrations at the population or species level, and can be a powerful approach to testing both long-standing and new questions in biology.

HormoneBase, a population-level database of steroid hormone levels across vertebrates.

Hormones are central regulators of organismal function and flexibility that mediate a diversity of phenotypic traits from early development through senescence. Yet despite these important roles, basic questions about how and why hormone systems vary within and across species remain unanswered. Here we describe HormoneBase, a database of circulating steroid hormone levels and their variation across vertebrates. This database aims to provide all available data on the mean, variation, and range of plasma glucocorticoids (both baseline and stress-induced) and androgens in free-living and un-manipulated adult vertebrates. HormoneBase (www.HormoneBase.org) currently includes >6,580 entries from 476 species, reported in 648 publications from 1967 to 2015, and unpublished datasets. Entries are associated with data on the species and population, sex, year and month of study, geographic coordinates, life history stage, method and latency of hormone sampling, and analysis technique. This novel resource could be used for analyses of the function and evolution of hormone systems, and the relationships between hormonal variation and a variety of processes including phenotypic variation, fitness, and species distributions.

Energetic costs of performance in trained and untrained Anolis carolinensis lizards.

The energetic costs of performance constitute a non-trivial component of animals' daily energetic budgets. However, we currently lack an understanding of how those costs are partitioned among the various stages of performance development, maintenance and production. We manipulated individual investment in performance by training Anolis carolinensis lizards for endurance or sprinting ability. We then measured energetic expenditure both at rest and immediately following exercise to test whether such training alters the maintenance and production costs of performance. Trained lizards had lower resting metabolic rates than controls, suggestive of a maintenance saving associated with enhanced performance as opposed to a cost. Production costs also differed, with sprint-trained lizards incurring a larger energetic performance cost and experiencing longer recovery times compared with endurance trained and control animals. Although performance training modifies metabolism, production costs are probably the key drivers of trade-offs between performance and other life-history traits in this species.