Does capacity to produce androgens underlie variation in female ornamentation and territoriality in White-shouldered Fairywren (Malurus alboscapulatus)?

Historic bias toward study of sex hormones and sexual ornamentation in males currently constrains our perspective of hormone-behavior-phenotype relationships. Resolving how ornamented female phenotypes evolve is particularly important for understanding the diversity of social signals across taxa. Studies of both males and females in taxa with variable female phenotypes are needed to establish whether sexes share mechanisms underlying expression of signaling phenotypes and behavior. White-shouldered Fairywren (Malurus alboscapulatus) subspecies vary in female ornamentation, baseline circulating androgens, and response to territorial intrusion. The moretoni ornamented female subspecies is characterized by higher female, but lower male baseline androgens, and a stronger pair territorial response relative to pairs from the lorentzi unornamented female subspecies. Here we address whether subspecific differences in female ornamentation, baseline androgens, and pair territoriality are associated with ability to elevate androgens following gonadotropin releasing hormone (GnRH) challenge and in response to simulated territorial intrusion. We find that subspecies do not differ in their capacity to produce androgens in either sex following GnRH or simulated territorial intrusion (STI) challenges. STI-induced androgens were predictive of degree of response to territorial intrusions in females only, but the direction of the effect was mixed. GnRH-induced androgens did not correlate with response to simulated intruders, nor did females sampled during intrusion elevate androgens relative to flushed controls, suggesting that increased androgens are not necessary for the expression of territorial defense behaviors. Collectively, our results suggest that capacity to produce androgens does not underlie subspecific patterns of female ornamentation, territoriality, and baseline plasma androgens.

Extended and cumulative effects of experimentally induced intergroup conflict in a cooperatively breeding mammal.

Conflict between rival groups is rife in nature. While recent work has begun exploring the behavioural consequences of this intergroup conflict, studies have primarily considered just the 1-2 h immediately after single interactions with rivals or their cues. Using a habituated population of wild dwarf mongooses (Helogale parvula), we conducted week-long manipulations to investigate longer-term impacts of intergroup conflict. Compared to a single presentation of control herbivore faeces, one rival-group faecal presentation (simulating a territorial intrusion) resulted in more within-group grooming the following day, beyond the likely period of conflict-induced stress. Repeated presentations of outsider cues led to further changes in baseline behaviour by the end of the week: compared to control weeks, mongooses spent less time foraging and foraged closer to their groupmates, even when there had been no recent simulated intrusion. Moreover, there was more baseline territorial scent-marking and a higher likelihood of group fissioning in intrusion weeks. Consequently, individuals gained less body mass at the end of weeks with repeated simulated intrusions. Our experimental findings provide evidence for longer-term, extended and cumulative, effects of an elevated intergroup threat, which may lead to fitness consequences and underpin this powerful selective pressure.

Who rises to the challenge? Testing the Challenge Hypothesis in fish, amphibians, reptiles, and mammals.

According to the Challenge Hypothesis, social interactions, particularly among males, have a strong influence on circulating androgen levels. Specifically, males should respond to social challenges from conspecific males with a rapid increase in plasma androgen levels which support and stimulate further aggression. This basic tenet of the Challenge Hypothesis, an androgen increase in response to a social challenge from another male, has been tested in all vertebrate classes. While early studies generally supported the Challenge Hypothesis, more recent work has noted numerous exceptions, particularly in birds. Here, we conduct a meta-analysis of studies in fish, amphibians, non-avian reptiles, and mammals that test the prediction that circulating androgen levels of males should increase in response to an experimental challenge from another male. We found that teleost fish often increase androgens during such challenges, but other vertebrate groups show more mixed results. Why should fish be different from the other taxa? In fish with paternal care of young, the potential conflict between mating, being aggressive towards other males, and taking care of offspring is alleviated, because females typically choose males based on their defense of an already existing nest. Hence, rather than regulating the trade-off between mating, aggression, and parenting, androgens may have been co-opted to promote all three behaviors. For other taxa, increasing androgen levels only makes sense when the increase directly enhances reproductive success. Thus, the increase in androgen levels is a response to mating opportunities rather than a response to challenge from another male. To further our understanding of the role of a change in androgen levels in mediating behavioral decision-making between mating, aggression, and parenting, we need studies that address the behavioral consequences of an increase in androgens after male-male encounters and studies that test the androgen responsiveness of species that differ in the degree of paternal care.

Baseline and stress-induced levels of corticosterone in male and female Afrotropical and European temperate stonechats during breeding.

BACKGROUND: Latitudinal variation in avian life histories falls along a slow-fast pace of life continuum: tropical species produce small clutches, but have a high survival probability, while in temperate species the opposite pattern is found. This study investigated whether differential investment into reproduction and survival of tropical and temperate species is paralleled by differences in the secretion of the vertebrate hormone corticosterone (CORT). Depending on circulating concentrations, CORT can both act as a metabolic (low to medium levels) and a stress hormone (high levels) and, thereby, influence reproductive decisions. Baseline and stress-induced CORT was measured across sequential stages of the breeding season in males and females of closely related taxa of stonechats (Saxicola spp) from a wide distribution area. We compared stonechats from 13 sites, representing Canary Islands, European temperate and East African tropical areas. Stonechats are highly seasonal breeders at all these sites, but vary between tropical and temperate regions with regard to reproductive investment and presumably also survival. RESULTS: In accordance with life-history theory, during parental stages, post-capture (baseline) CORT was overall lower in tropical than in temperate stonechats. However, during mating stages, tropical males had elevated post-capture (baseline) CORT concentrations, which did not differ from those of temperate males. Female and male mates of a pair showed correlated levels of post-capture CORT when sampled after simulated territorial intrusions. In contrast to the hypothesis that species with low reproduction and high annual survival should be more risk-sensitive, tropical stonechats had lower stress-induced CORT concentrations than temperate stonechats. We also found relatively high post-capture (baseline) and stress-induced CORT concentrations, in slow-paced Canary Islands stonechats. CONCLUSIONS: Our data support and refine the view that baseline CORT facilitates energetically demanding activities in males and females and reflects investment into reproduction. Low parental workload was associated with lower post-capture (baseline) CORT as expected for a slow pace of life in tropical species. On a finer resolution, however, this tropical-temperate contrast did not generally hold. Post-capture (baseline) CORT was higher during mating stages in particular in tropical males, possibly to support the energetic needs of mate-guarding. Counter to predictions based on life history theory, our data do not confirm the hypothesis that long-lived tropical populations have higher stress-induced CORT concentrations than short-lived temperate populations. Instead, in the predator-rich tropical environments of African stonechats, a dampened stress response during parental stages may increase survival probabilities of young. Overall our data further support an association between life history and baseline CORT, but challenge the role of stress-induced CORT as a mediator of tropical-temperate variation in life history.

Territorial aggression does not feed back on testosterone in a multiple-brooded songbird species with breeding and non-breeding season territoriality, the European stonechat.

Testosterone mediates reproductive behaviours in male vertebrates. For example, breeding season territoriality depends on testosterone in many species of birds and in some, territorial interactions feed back on testosterone concentrations. However, the degree to which territorial behaviour and testosterone are associated differs even between species with seemingly similar life histories, especially between species that also defend territories outside the breeding season. Here, we investigate the link between territorial behaviour and testosterone in European stonechats. Previous studies found that territorial aggression in stonechats depends on testosterone in a breeding, but not in a non-breeding context. We investigated whether stonechats show a rise in testosterone during simulated territorial intrusions (STI) during the breeding season. Post-capture testosterone concentrations of males caught after an STI were not higher than those of males caught in a control situation regardless of breeding stage. However, most of the males would have been able to mount a testosterone response because the same individuals that did not increase testosterone during the STI showed a substantial increase in testosterone after injections of gonadotropin-releasing hormone (GnRH). GnRH-induced and post-capture testosterone concentrations were positively correlated and both decreased with successive breeding stages. Further, territory owners with a short latency to attack the decoy expressed higher post-capture testosterone concentrations than males with a longer latency to attack the decoy. Thus, there is no evidence for behavioural feedback on testosterone concentrations during male-male interactions in stonechats. In combination with previous studies our data suggest that testosterone functions as an on/off switch of high intensity territorial aggression during the breeding season in stonechats. The among-species variation in the androgen control of territorial behaviour may be only partly a result of environmental differences. Instead, potential differences in how territoriality evolved in different species may have influenced whether and how a reproductive hormone such as testosterone was co-opted into the mechanistic control of territorial behaviour.

Intra- and interspecific social challenges modulate the levels of an androgen precursor in a seasonally territorial tropical damselfish.

Studies on different vertebrate groups have provided evidence that androgen levels in males increase after competitive social interactions during the breeding season, as postulated by the Challenge Hypothesis. However, social modulation of androgen levels may vary with latitude and may differ between species holding seasonal versus year-round territories. Here, we tested the Challenge Hypothesis on a seasonal tropical damselfish, Abudefduf sexfasciatus, where males temporarily defend territory and eggs against both intra- and interspecific individuals. Carrying out simulated territorial intrusions (STIs) in the laboratory, we document for the first time a consistent increase in the plasma level of the androgen precursor 11-ketoandrostenedione (11KA) in fish confronted to either intra- or interspecific challenges. Collecting samples in the field also revealed higher 11KA levels in fish facing frequent territorial interactions than in non-territorial individuals. Levels of 11-ketotestosterone (11KT) were high in territorial males in the field, but were not incremented after simulated territorial intrusions in the laboratory. Plasma levels of cortisol and testosterone were not affected by challenges but were different in wild and captive specimens. Although the endocrine responses to STIs did not differ between intra- and interspecific challenges, agonistic displays expressed by resident fish were more intense towards intraspecific intruders. Taken together, our study emphasizes the need to incorporate androgen precursor concentrations to advance our understanding on the physiology of territorial interactions.

Behavioural response of a migratory songbird to geographic variation in song and morphology.

INTRODUCTION: Sexually selected traits contribute substantially to evolutionary diversification, for example by promoting assortative mating. The contributing traits and their relevance for reproductive isolation differ between species. In birds, sexually selected acoustic and visual signals often undergo geographic divergence. Clines in these phenotypes may be used by both sexes in the context of sexual selection and territoriality. The ways conspecifics respond to geographic variation in phenotypes can give insights to possible behavioural barriers, but these may depend on migratory behaviour. We studied a migratory songbird, the Stonechat, and tested its responsiveness to geographic variation in male song and morphology. The traits are acquired differently, with possible implications for population divergence. Song can evolve quickly through cultural transmission, and thus may contribute more to the establishment of geographic variation than inherited morphological traits. We first quantified the diversity of song traits from different populations. We then tested the responses of free-living Stonechats of both sexes to male phenotype with playbacks and decoys, representing local and foreign stimuli derived from a range of distances from the local population. RESULTS: Both sexes discriminated consistently between stimuli from different populations, responding more strongly to acoustic and morphological traits of local than foreign stimuli. Time to approach increased, and time spent close to the stimuli and number of tail flips decreased consistently with geographic distance of the stimulus from the local population. Discriminatory response behaviour was more consistent for acoustic than for morphological traits. Song traits of the local population differed significantly from those of other populations. CONCLUSIONS: Evaluating an individual's perception of geographic variation in sexually selected traits is a crucial first step for understanding reproductive isolation mechanisms. We have demonstrated that in both sexes of Stonechats the responsiveness to acoustic and visual signals decreased with increasing geographic distance of stimulus origin. These findings confirm consistent, fine discrimination for both learned song and inherited morphological traits in these migratory birds. Maintenance or further divergence in phenotypic traits could lead to assortative mating, reproductive isolation, and potentially speciation.

Photoperiod modulation of aggressive behavior is independent of androgens in a tropical cichlid fish.

Photoperiod is a major environmental cue that signals breeding conditions in animals living in temperate climates. Therefore, the activity of the reproductive (i.e. hypothalamic-pituitary-gonadal, HPG) axis and of the expression of reproductive behaviors, including territoriality, is responsive to changes in day length. However, at low latitudes the seasonal variation in day length decreases dramatically and photoperiod becomes less reliable as a breeding entraining cue in tropical species. In spite of this, some tropical mammals and birds have been found to still respond to small amplitude changes in photoperiod (e.g. 17min). Here we tested the effect of 2 photoperiod regimes, referred to as long-day (LD: 16L:08D) and short-day (SD: 08L:16D), on the activity of the HPG axis, on aggressive behavior and in the androgen response to social challenges in males of the tropical cichlid fish Tilapia rendalli. For each treatment, fish were transferred from a pre-treatment photoperiod of 12L:12D to their treatment photoperiod (either LD or SD) in which they were kept for 20days on stock tanks. Afterwards, males were isolated for 4days in glass aquaria in order to establish territories and initial androgen levels (testosterone, T; 11-ketotestosterone, KT) were assessed. On the 4th day, territorial intrusions were promoted such that 1/3 of the isolated males acted as residents and another 1/3 as intruders. Territorial intrusions lasted for 1h to test the effects of a social challenge under different photoperiod regimes. Photoperiod treatment (either SD or LD) failed to induce significant changes in the HPG activity, as measured by androgen levels and gonadosomatic index. However, SD increased the intensity of aggressive behaviors and shortened the time to settle a dominance hierarchy in an androgen-independent manner. The androgen responsiveness to the simulated territorial intrusion was only present in KT but not for T. The percent change in KT levels in response to the social challenge was different between treatments (SD>LD) and between male types (resident>intruder). The higher androgen response to a social challenge in residents under SD may be explained by the time course of the androgen response that due to the long time it takes to fight resolution under LD, might have been delayed. This result illustrates the importance of incorporating time response data in social endocrinology studies.

Regulation of plasma testosterone, corticosterone, and metabolites in response to stress, reproductive stage, and social challenges in a desert male songbird.

In many male vertebrates, the secretion of reproductive (gonadal androgens) and adrenocortical (glucocorticoids) hormones varies seasonally and in response to environmental stimuli, and these hormones exert numerous behavioral and metabolic effects. We performed two field studies on adult male Rufous-winged Sparrows, Peucaea carpalis, a Sonoran Desert rain-dependent sedentary species, to (a) determine seasonal changes in initial (baseline) and acute stress-induced plasma testosterone (T), corticosterone (CORT), and two metabolites (uric acid and glucose) and (b) compare the effects of two types of social challenge (song playback or simulated territorial intrusion consisting of song playback plus exposure to a live decoy bird) on plasma T, CORT, these metabolites, and territorial behavior. Initial plasma T was higher during the summer breeding period than during post-breeding molt. Acute stress resulting from capture and restraint for 30 min decreased plasma T in breeding condition birds but not in the fall, revealing that this decrease is seasonally regulated. Initial plasma CORT did not change seasonally, but plasma CORT increased in response to acute stress. This increase was likewise seasonally regulated, being relatively smaller during autumnal molt than in the summer. We found no evidence that acute stress levels of CORT are functionally related to stress-depressed plasma T and, therefore, that plasma T decreases during stress as a result of elevated plasma CORT. Thirty minutes of exposure to simulated territorial intrusion resulted in different behavior than 30 min of exposure to song playback, with increased time spent near the decoy and decreased number of overhead flights. Neither type of social challenge influenced plasma T, thus offering no support for the hypothesis that plasma T either responds to or mediates the behavioral effects of social challenge. Exposure to both social challenges elevated plasma CORT, but simulated territorial intrusion was more effective in this respect than song playback. Plasma uric acid and glucose decreased during acute stress, but only plasma uric acid decreased during social challenge. Thus, an elevation in plasma CORT was consistently associated with a decrease in plasma uric acid, but not with a change in glycemia. These results enhance our understanding of the short-term relationships between T, CORT, and avian territorial behavior. They provide novel information on the endocrine effects of acute stress, in particular on plasma T, in free-ranging birds, and are among the first in these birds to link these effects to metabolic changes.

Highly context-specific activation of the HPG axis in the dark-eyed junco and implications for the challenge hypothesis.

One of the best studied hormone-behavior interactions is the transient rise in testosterone (T) associated with male-male aggression. However, recent research on songbirds has demonstrated numerous exceptions to this pattern.One species previously thought to elevate T in response to a simulated territorial intrusion is the dark-eyed junco (Junco hyemalis). Here, we show that under most circumstances male juncos do not elevate circulating T or CORT levels in response to social stimuli, despite being physiologically capable of elevating T as indicated by their response to GnRH. The lack of hormonal response was found regardless of the sex of the social stimulus (singing male vs. soliciting female), its sensory modality (song only, song + live lure, song + taxidermic mount), or the timecourse of sampling. Notably, males did elevate T levels when exposed to a simulated territorial intrusion in the days following simulated predation of their chicks. Whether the high T seen in these narrow circumstances represents stage-dependent social modulation of T or re-activation of male reproductive physiology in preparation for re-nesting (i.e. socially independent T modulation) remains to be determined. It is clear, however, that activation of the HPG axis is highly context-specific for male juncos. These results highlight important and unresolved issues regarding the socially mediated component of the challenge hypothesis and how it relates to the evolution of hormone-mediated traits.

Females Are More Aggressive Than Males towards Same- and Opposite-Sex Intruders in the Blue Tit (Cyanistes caeruleus).

During the breeding season, aggression is expressed to gain access to resources such as territories and mates and protect offspring. Female aggressiveness has received much less attention than male aggressiveness, and few studies have examined female and male aggressiveness towards intruders of both sexes in the same species. We compared female and male aggressiveness towards same- and opposite-sex intruders during the egg-laying period in blue tits (Cyanistes caeruleus) using simulated territorial intrusions. For each sex, we examined the occurrence of different behavioural responses during agonistic encounters, and compared the intensity and individual consistency of intra- and inter-sexual aggression using same- and opposite-sex taxidermy mounts. Our results show that females are the more aggressive sex. Both sexes showed similar behaviours during simulated intrusions, although females were never observed singing and males never entered the nest box. In females, aggression was predominantly independent of the sex of the intruder, while males sang more from a distance during male-male encounters. The relative levels of aggression (pecking and perching on the mounts) during intra- and intersexual conflicts were consistent for females, but not for males. Females might be under stronger selection for aggressive phenotypes due to nest-hole competition and larger reproductive investments.

Bidirectional relationships between testosterone and aggression: a critical analysis of four predictions.

Experimentally elevated testosterone (T) often leads to enhanced aggression, with examples across many different species, including both males and females. Indeed, the relationship between T and aggression is among the most well-studied and fruitful areas of research at the intersection of behavioral ecology and endocrinology. This relationship is also hypothesized to be bidirectional (i.e., T influences aggression, and aggression influences T), leading to four key predictions: (1) Individuals with higher T levels are more aggressive than individuals with lower T. (2) Seasonal changes in aggression mirror seasonal changes in T secretion. (3) Aggressive territorial interactions stimulate increased T secretion. (4) Temporary elevations in T temporarily increase aggressiveness. These predictions cover a range of timescales, from a single snapshot in time, to rapid fluctuations, and to changes over seasonal timescales. Adding further complexity, most predictions can also be addressed by comparing among individuals or with repeated sampling within-individuals. In our review, we explore how the spectrum of results across predictions shapes our understanding of the relationship between T and aggression. In all cases, we can find examples of results that do not support the initial predictions. In particular, we find that predictions 1-3 have been tested frequently, especially using an among-individual approach. We find qualitative support for all three predictions, though there are also many studies that do not support predictions 1 and 3 in particular. Prediction 4, on the other hand, is something that we identify as a core underlying assumption of past work on the topic, but one that has rarely been directly tested. We propose that when relationships between T and aggression are individual-specific or condition-dependent, then positive correlations between the two variables may be obscured or reversed. In essence, even though T can influence aggression, many assumed or predicted relationships between the two variables may not manifest. Moving forward, we urge greater attention to understanding how and why it is that these bidirectional relationships between T and aggression may vary among timescales and among individuals. In doing so, we will move towards a deeper understanding on the role of hormones in behavioral adaptation.

Testing hormonal responses to real and simulated social challenges in a competitive female bird.

Competitive interactions often occur in series; therefore animals may respond to social challenges in ways that prepare them for success in future conflict. Changes in the production of the steroid hormone testosterone (T) are thought to mediate phenotypic responses to competition, but research over the past few decades has yielded mixed results, leading to several potential explanations as to why T does not always elevate following a social challenge. Here, we measured T levels in tree swallows (Tachycineta bicolor), a system in which females compete for limited nesting cavities and female aggression is at least partially mediated by T. We experimentally induced social challenges in two ways: (1) using decoys to simulate territorial intrusions and (2) removing subsets of nesting cavities to increase competition among displaced and territory-holding females. Critically, these experiments occurred pre-laying, when females are physiologically capable of rapidly increasing circulating T levels. However, despite marked aggression in both experiments, T did not elevate following real or simulated social challenges, and in some cases, socially challenged females had lower T levels than controls. Likewise, the degree of aggression was negatively correlated with T levels following a simulated territorial intrusion. Though not in line with the idea that social challenges prompt T elevation in preparation for future challenges, these patterns nevertheless connect T to territorial aggression in females. Coupled with past work showing that T promotes aggression, these results suggest that T may act rapidly to allow animals to adaptively respond to the urgent demands of a competitive event.

Female aggression towards same-sex rivals depends on context in a tropical songbird.

Agonistic conflict is ubiquitous throughout taxa, although the intensity of aggression observed is often highly variable across contexts. For socially monogamous species, a coordinated effort by both pair members can improve the chances of successfully warding off challengers and reinforce pair bonds. However, the intensity of aggression exerted by any one pair member may vary with respect to contextual factors, including the intensity of their mate's aggression. Thus, experimentally exploring how individuals respond to potential rivals via multiple assays with varying social contexts can advance our basic understanding of how aggression varies in socially monogamous systems. We used simulated territorial intrusion and mirror image simulation assays to explore this issue in white-shouldered fairywrens (Malurus alboscapulatus moretoni) of Papua New Guinea. While males tended to be more responsive than females during simulated territorial intrusions, females were more aggressive towards their mirrored reflection than males. Further, individual females that were most aggressive in mirror image simulations were the least aggressive during simulated territorial intrusions, whereas males were inconsistent. These results suggest that female behavioral phenotypes appear to be flexible, relative to context. We discuss how multiple commonly used measurements of aggression might in fact measure different types of responses.

Nest construction and presence do not alter territorial aggression in male threespine stickleback.

Constructing a nest within a mating territory provides a clear benefit to the resident, particularly by improving the opportunity to mate. It is unclear whether animals who use nests exclusively for reproductive purposes account for either the effort invested or the resulting increase in mating potential when valuing their territories. We sought to explicitly reveal a nest's added subjective resource value through within-group comparison of aggressive behaviour before and after nest construction. An increase in aggression following construction would indicate that the resident perceives greater subjective value in his territory, and thus values the nest. Threespine stickleback, Gasterosteus aculeatus, fish demonstrate stereotypical aggressive behaviours during an easily induced territorial defence. The male's nest is used exclusively for reproductive purposes, avoiding any confound of shelter. Contrary to our hypothesis, neither nest presence, timing of construction, nor nesting outcome was associated with differences in behavioural measures of territorial aggression. Assessed behaviours were robust, repeatable and inter-correlated. We conclude that territorial aggression is neither predictive of nor altered by nesting in threespine stickleback fish. Our results suggest that nests used transiently for a portion of the mating season add negligible subjective resource value to a territory. We additionally demonstrate that examinations of territorial aggression in sticklebacks do not need to control for nest building, improving statistical power by decreasing dropout rates. These results dovetail with recent work in other fish species to suggest that assessment of territorial aggression absent a nest may be practicable for fish in general.

Species partitioning in a temperate mountain chain: Segregation by habitat vs. interspecific competition.

Disentangling the relative influence of the environment and biotic interactions in determining species coexistence patterns is a major challenge in ecology. The zonation occurring along elevation gradients, or at bioclimatic contact zones, offers a good opportunity to improve such understanding because the small scale at which the partitioning occurs facilitates inference based on experiments and ecological modelling. We studied the influence of abiotic gradients, habitat types, and interspecific competition in determining the spatial turnover between two pipit and two bunting species in NW Spain. We explored two independent lines of evidence to draw inference about the relative importance of environment and biotic interactions in driving range partitioning along elevation, latitude, and longitude. We combined occurrence data with environmental data to develop joint species distribution models (JSDM), in order to attribute co-occurrence (or exclusion) to shared (or divergent) environmental responses and to interactions (attraction or exclusion). In the same region, we tested for interference competition by means of playback experiments in the contact zone. The JSDMs highlighted different responses for the two species pairs, although we did not find direct evidence of interspecific aggressiveness in our playback experiments. In pipits, partitioning was explained by divergent climate and habitat requirements and also by the negative correlations between species not explained by the environment. This significant residual correlation may reflect forms of competition others than direct interference, although we could not completely exclude the influence of unmeasured environmental predictors. When bunting species co-occurred, it was because of shared habitat preferences, and a possible limitation to dispersal might cause their partitioning. Our results indicate that no single mechanism dominates in driving the distribution of our study species, but rather distributions are determined by the combination of many small forces including biotic and abiotic determinants of niche, whose relative strengths varied among species.

Does a short-term increase in testosterone affect the intensity or persistence of territorial aggression? - An approach using an individual's hormonal reactive scope to study hormonal effects on behavior.

In this study, we describe an approach based on an individual's hormonal reactive scope to study short-term effects of hormones on behavior. The control of territorial aggression has been traditionally linked to testosterone. Males of some vertebrate species show an increase in testosterone during territorial interactions and implantation studies suggest that such an increase in testosterone enhances the intensity and persistence of aggression. Here, we tested whether a short-term maximum release of testosterone - based on an individual's hormonal reactive scope - affects the intensity or persistence of territorial aggression in male black redstarts, a bird species in which testosterone does not increase during territorial encounters. An injection with gonadotropin-releasing-hormone (GnRH) induced a physiological peak in plasma testosterone that was specific for each individual (=individual reactive scope). However, such short-term surges in an individual's testosterone concentration did not affect the intensity or persistence of aggression. In conclusion, this study demonstrated (1) that a species that naturally does not increase testosterone during male-male encounters would not benefit from such an increase in terms of being more aggressive, (2) that behavioral studies using GnRH-injections represent a promising approach to study species differences in androgen responsiveness, and (3) that injections of releasing or tropic hormones in general may be a suitable approach to study short-term influences of hormones on behavior. These injections effectively mimic the potential short-term changes in hormones that can occur in the real life of individuals and enable us to study the effects of hormonal changes on behavior or other traits within an ecological and evolutionary framework.