Social status influences relationships between hormones and oxidative stress in a cichlid fish.

An individual's social environment can have widespread effects on their physiology, including effects on oxidative stress and hormone levels. Many studies have suggested that variation in oxidative stress experienced by individuals of different social statuses might be due to endocrine differences, however, few studies have evaluated this hypothesis. Here, we assessed whether a suite of markers associated with oxidative stress in different tissues (blood/plasma, liver, and gonads) had social status-specific relationships with circulating testosterone or cortisol levels in males of a cichlid fish, Astatotilapia burtoni. Across all fish, blood DNA damage (a global marker of oxidative stress) and gonadal synthesis of reactive oxygen species [as indicated by NADPH-oxidase (NOX) activity] were lower when testosterone was high. However, high DNA damage in both the blood and gonads was associated with high cortisol in subordinates, but low cortisol in dominants. Additionally, high cortisol was associated with greater production of reactive oxygen species (greater NOX activity) in both the gonads (dominants only) and liver (dominants and subordinates). In general, high testosterone was associated with lower oxidative stress across both social statuses, whereas high cortisol was associated with lower oxidative stress in dominants and higher oxidative stress in subordinates. Taken together, our results show that differences in the social environment can lead to contrasting relationships between hormones and oxidative stress.

Mothers modify the cost of reproduction by dynamic changes in antioxidant function and filial cannibalism.

Investment in current reproduction may negatively influence subsequent fitness. Oxidative stress has been proposed as a potential mediator of this trade-off between current and future reproductive success. However, evidence of reproduction causing oxidative stress is limited, possibly owing to compensatory mechanisms that counteract oxidative insults. Here we test the idea that organisms protect against oxidative challenges through a dynamic interaction between behavioural and physiological adjustments at different stages of reproduction. To test this idea, we manipulated maternal care in the mouthbrooding cichlid fish Astatotilapia burtoni by allowing females to continue care (brooders) or by preventing care (non-brooders). We found that brooders depleted the pool of antioxidants as brood care progressed; however, we only observed increased oxidative DNA damage at the early stage of care relative to non-brooders, possibly owing to upregulated antioxidant protection during later stages of care. Most brooders adjusted parental investment by consuming some of their offspring during mouthbrooding. Intriguingly, the level of filial cannibalism was positively related to liver antioxidant function. These changes in antioxidant function and filial cannibalism allow parents to manage the cost of reproduction and are important for our understanding of how oxidative stress mediates life-history trade-offs.

Social dominance and reproduction result in increased integration of oxidative state in males of an African cichlid fish.

Oxidative stress is a potential cost of social dominance and reproduction, which could mediate life history trade-offs between current and future reproductive fitness. However, the evidence for an oxidative cost of social dominance and reproduction is mixed, in part because organisms have efficient protective mechanisms that can counteract oxidative insults. Further, previous studies have shown that different aspects of oxidative balance, including oxidative damage and antioxidant function, vary dramatically between tissue types, yet few studies have investigated oxidative cost in terms of interconnectedness and coordination within the system. Here, we tested whether dominant and subordinate males of the cichlid Astatotilapia burtoni differ in integration of different components of oxidative stress. We assessed 7 markers of oxidative stress, which included both oxidative damage and antioxidant function in various tissue types (total of 14 measurements). Across all oxidative stress measurements, we found more co-regulated clusters in dominant males, suggesting that components of oxidative state are more functionally integrated in dominant males than they are in subordinate males. We discuss how a high degree of functional integration reflects increased robustness or efficiency of the system (e.g. increased effectiveness of antioxidant machinery in reducing oxidative damage), but we also highlight potential costs (e.g. activation of cytoprotective mechanisms may have unwanted pleiotropic effects). Overall, our results suggest that quantifying the extent of functional integration across different components of oxidative stress could reveal insights into the oxidative cost of important life history events.

Antioxidant capacity differs across social ranks and with ascension in males of a group-living fish.

Animals that live in groups often form hierarchies in which an individual's behaviour and physiology varies based on their social rank. Occasionally, a subordinate can ascend into a dominant position and the ascending individual must make rapid behavioural and physiological adjustments to solidify their dominance. These periods of social transition and instability can be stressful and ascending individuals often incur large metabolic costs that could influence their oxidative status. Most previous investigations examining the link between oxidative status and the social environment have done so under stable social conditions and have evaluated oxidative status in a single tissue. Therefore, evaluations of how oxidative status is regulated across multiple tissues during periods of social flux would greatly enhance our understanding of the relationship between oxidative status and the social environment. Here, we assessed how antioxidant capacity in three tissues (brain, gonad, and muscle) varied among dominant, subordinate, and ascending males of the group-living cichlid fish, Neolamprologus pulcher. Antioxidant capacity in the brain and muscle of ascending males was intermediate to that of dominant (highest levels) and subordinate males (lowest levels) and correlated with differences in social and locomotor behaviours, respectively. Gonad antioxidant capacity was lower in ascending males than in dominant males. However, gonad antioxidant capacity was positively correlated with the size of ascending males' gonads suggesting that ascending males may increase gonad antioxidant capacity as they develop their gonads. Overall, our results highlight the widespread physiological consequences of social ascension and emphasize the importance of tissue-specific measures of oxidative status.

Social instability influences rank-specific patterns of oxidative stress in a cichlid fish.

In many animal societies, dominant individuals have priority access to resources. However, defending high rank can be costly, especially in unstable social hierarchies where there is more intense competition. Oxidative stress has been proposed as a potential cost of social dominance, but few studies have examined this cost in relation to social stability. We studied the cost of social dominance in the cichlid fish Astatotilapia burtoni by manipulating social stability among males in replicate naturalistic communities for 22 weeks. We found that our social stability treatment influenced status-specific patterns in 3 out of 6 measurements of oxidative stress. Specifically, dominant males experienced increased plasma oxidative damage (measured as reactive oxygen metabolites, ROMs) compared with subordinate males in stable hierarchies only. Subordinate males in unstable hierarchies had higher ROMs than their stable community counterparts, but we found no effect of social stability treatment for dominant males. However, dominant males tended to have reduced total antioxidant capacity (TAC) in the liver when compared with subordinate males in unstable hierarchies, suggesting that the cost of social dominance is higher in unstable hierarchies. There were no effects of status and treatment on gonad TAC, muscle TAC or oxidative DNA damage. We conclude that the stability of the social environment influences the relative cost of social dominance in a tissue- and marker-specific manner.

Social network dynamics predict hormone levels and behavior in a highly social cichlid fish.

Group living confers many benefits while simultaneously exposing group members to intense competition. An individual's rise to prominence within a group may conflict with the overall functioning of the group. There is therefore a complex and dynamic relationship between the behavioral displays that directly benefit an individual, the consequences of these actions for the community, and how they feed back on individual-level fitness. We used a network analysis approach to study the link between behavior, social stability, and steroid hormone levels in replicate communities of the cichlid fish, Astatotilapia burtoni, which live in social groups with a dominance hierarchy. We demonstrate that individual behavior can have direct and indirect effects on the behavior of others while also affecting group characteristics. Our results show that A. burtoni males form stable social networks, where dominant individuals act as hubs for social interactions. However, there was variation in the temporal stability in these networks, and this variation in stability impacted hormone levels. Dominant males had higher testosterone levels, however, the differences in testosterone levels between dominant and subordinate males were greatest in stable communities. In sum, our analyses provide novel insights into the processes by which individual and community properties interact.

How does male-male competition generate negative frequency-dependent selection and disruptive selection during speciation?

Natural selection has been shown to drive population differentiation and speciation. The role of sexual selection in this process is controversial; however, most of the work has centered on mate choice while the role of male-male competition in speciation is relatively understudied. Here, we outline how male-male competition can be a source of diversifying selection on male competitive phenotypes, and how this can contribute to the evolution of reproductive isolation. We highlight how negative frequency-dependent selection (advantage of rare phenotype arising from stronger male-male competition between similar male phenotypes compared with dissimilar male phenotypes) and disruptive selection (advantage of extreme phenotypes) drives the evolution of diversity in competitive traits such as weapon size, nuptial coloration, or aggressiveness. We underscore that male-male competition interacts with other life-history functions and that variable male competitive phenotypes may represent alternative adaptive options. In addition to competition for mates, aggressive interference competition for ecological resources can exert selection on competitor signals. We call for a better integration of male-male competition with ecological interference competition since both can influence the process of speciation via comparable but distinct mechanisms. Altogether, we present a more comprehensive framework for studying the role of male-male competition in speciation, and emphasize the need for better integration of insights gained from other fields studying the evolutionary, behavioral, and physiological consequences of agonistic interactions.

Does Blue Uniform Color Enhance Winning Probability in Judo Contests?

The color of an athlete's uniform may have an effect on psychological functioning and consequently bias the chances of winning contests in sport competition. Several studies reported a winning bias for judo athletes wearing a blue outfit relative to those wearing a white outfit. However, we argue there is no winning bias and that previous studies were confounded and based on small and specific data sets. We tested whether blue biases winning in judo using a very extensive judo data set (45,874 contests from all international judo tournaments between 2008 and 2014). In judo, the first called athlete for the fight used to wear the blue judogi but this was changed to the white judogi in 2011. This switch enabled us to compare the win bias before and after this change to isolate the effect of the color of the judogi. We found a significant win bias for the first called athlete, but this effect was not significantly related to the color of the judogi. The lack of a significant win effect of judogi color suggests that blue does not bias winning in judo, and that the blue-white pairing ensures an equal level of play. Our study shows the importance of thoroughly considering alternative explanations and using extensive datasets in color research in sports and psychology.

The melanocortin system regulates body pigmentation and social behaviour in a colour polymorphic cichlid fish.

The melanocortin system is a neuroendocrine system that regulates a range of physiological and behavioural processes. We examined the extent to which the melanocortin system simultaneously regulates colour and behaviour in the cichlid fish Astatotilapia burtoni We found that yellow males are more aggressive than blue males, in line with previous studies. We then found that exogenous α-melanocyte-stimulating hormone (α-MSH) increases yellowness of the body and dispersal of xanthophore pigments in both morphs. However, α-MSH had a morph-specific effect on aggression, with only blue males showing an increase in the rate of aggression. Exogenous agouti signalling peptide (ASIP), a melanocortin antagonist, did not affect coloration but reduced the rate of aggression in both colour morphs. Blue males had higher cortisol levels than yellow males. Neural gene expression of melanocortin receptors (mcr) and ligands was not differentially regulated between colour morphs. In the skin, however, mc1r and pro-opiomelanocortin (pomc) ß were upregulated in blue males, while asip 1 was upregulated in yellow males. The effects of α-MSH on behaviour and body coloration, combined with morph-specific regulation of the stress response and the melanocortin system, suggest that the melanocortin system contributes to the polymorphism in behaviour and coloration in A. burtoni.

Complex structural and regulatory evolution of the pro-opiomelanocortin gene family.

The melanocortin system is a neuroendocrine machinery that has been associated with phenotypic diversification in a number of vertebrate lineages. Central to the highly pleiotropic melanocortin system is the pro-opiomelanocortin (pomc) gene family, a family of pre-prohormones that each give rise to melanocyte stimulating hormone (MSH), adrenocorticotropic releasing hormone (ACTH), ß-lipotropin hormone, and ß-endorphin. Here we examine the structure, tissue expression profile, and pattern of cis transcriptional regulation of the three pomc paralogs (α1, α2, and ß) in the model cichlid fish Astatotilapia burtoni and other cichlids, teleosts, and mammals. We found that the hormone-encoding regions of pomc α1, pomc α2 and pomc ß are highly conserved, with a few notable exceptions. Surprisingly, the pomc ß gene of cichlids and pomacentrids (damselfish) encodes a novel melanocortin peptide, ε-MSH, as a result of a tandem duplication of the segment encoding ACTH. All three genes are expressed in the brain and peripheral tissues, but pomc α1 and α2 show a more spatially restricted expression profile than pomc ß. In addition, the promoters of each pomc gene have diverged in nucleotide sequence, which may have facilitated the diverse tissue-specific expression profiles of these paralogs across species. Increased understanding of the mechanisms regulating pomc gene expression will be invaluable to the study of pomc in the context of phenotypic evolution.

Immune activation is inversely related to, but does not cause variation in androgen levels in a cichlid fish species.

Studies on birds and mammals indicate that sexual traits may signal superior health because active immunity, like inflammatory responses to infections, is suppressive to the production of androgens that facilitate the expression of these traits. Here we test this possible pathway for honest signaling in a teleost species, Sarotherodon galilaeus, by activating the immune system with sheep red blood cells (SRBC), which is a non-pathogenic T- and B-cell stimulating antigen. Two weeks after the start of treatment adult males injected with SRBC showed a significant increase in antibody production in comparison with control males. The variation in specific antibody production was negatively related with variation in both testosterone and 11-ketotestosterone levels. This suggests that investment in immune protection is incompatible with increased activity of the hypothalamic-pituitary-gonadal axis. However, opposite to our expectation no difference in androgen levels was found between placebo and SRBC treatment suggesting that immune activation did not cause androgen suppression in our studied species.

Divergent hormonal responses to social competition in closely related species of haplochromine cichlid fish.

The diverse cichlid species flocks of the East African lakes provide a classical example of adaptive radiation. Territorial aggression is thought to influence the evolution of phenotypic diversity in this system. Most vertebrates mount hormonal (androgen, glucocorticoid) responses to a territorial challenge. These hormones, in turn, influence behavior and multiple aspects of physiology and morphology. Examining variation in competition-induced hormone secretion patterns is thus fundamental to an understanding of the mechanisms of phenotypic diversification. We test here the hypothesis that diversification in male aggression has been accompanied by differentiation in steroid hormone levels. We studied two pairs of sibling species from Lake Victoria belonging to the genera Pundamilia and Mbipia. The two genera are ecologically differentiated, while sibling species pairs differ mainly in male color patterns. We found that aggression directed toward conspecific males varied between species and across genera: Pundamilia nyererei males were more aggressive than Pundamilia pundamilia males, and Mbipia mbipi males were more aggressive than Mbipia lutea males. Males of both genera exhibited comparable attack rates during acute exposure to a novel conspecific intruder, while Mbipia males were more aggressive than Pundamilia males during continuous exposure to a conspecific rival, consistent with the genus difference in feeding ecology. Variation in aggressiveness between genera, but not between sibling species, was reflected in androgen levels. We further found that M. mbipi displayed lower levels of cortisol than M. lutea. Our results suggest that concerted divergence in hormones and behavior might play an important role in the rapid speciation of cichlid fishes.

'Winner effect' without winning: unresolved social conflicts increase the probability of winning a subsequent contest in a cichlid fish.

Previous winning experience increases the probability of winning a subsequent contest. However, it is not clear whether winning probability is affected only by the outcome of the contest (winning or losing) or whether fighting experience itself is also sufficient to induce this effect. We investigated this question in the East African cichlid fish Pundamilia spec. To create an unresolved conflict we allowed males to fight their own mirror image prior to a real fight against a size-matched non-mirror-stimulated control male. When males fight their own mirror image, the image's response corresponds to the action of the focal animal, creating symmetrical fighting conditions without the experience of losing or winning. We found that mirror-stimulated males were more likely to win an ensuing contest than control males. Interestingly, in this species mirror stimulation also induced an increase in circulating androgens, which is consistent with the hypothesis that stimulation of these sex steroids during aggressive encounters may prepare the animal for subsequent encounters. Our results suggest that fighting experience alone coupled with an androgen response, increases the likelihood of winning, even in the absence of a winning experience.

Male-male competition as a force in evolutionary diversification: evidence in haplochromine cichlid fish.

It has been suggested that intrasexual competition can be a source of negative frequency-dependent selection, causing agonistic character displacement and facilitating speciation and coexistence of (sibling) species. In this paper we synthesise the evidence that male-male and female-female competition contributes to cichlid diversification, showing that competition is stronger among same-coloured individuals than those with different colours. We argue that intrasexual selection is more complex because there are several examples where males do not bias aggression towards their own type. In addition, sibling species or colour morphs often show asymmetric dominance relationships. We briefly discuss potential mechanisms that might promote the maintenance of covariance between colour and aggression-related traits even in the face of gene-flow. We close by proposing several avenues for future studies that might shed more light on the role of intrasexual competition in cichlid diversification.

Frequency-dependent social dominance in a color polymorphic cichlid fish.

A mechanism commonly suggested to explain the persistence of color polymorphisms in animals is negative frequency-dependent selection. It could result from a social dominance advantage to rare morphs. We tested for this in males of red and blue color morphs of the Lake Victoria cichlid, Pundamilia. Earlier work has shown that males preferentially attack the males of their own morph, while red males are more likely to win dyadic contests with blue males. In order to study the potential contribution of both factors to the morph co-existence, we manipulated the proportion of red and blue males in experimental assemblages and studied its effect on social dominance. We then tried to disentangle the effects of the own-morph attack bias and social dominance of red using simulations. In the experiment, we found that red males were indeed socially dominant to the blue ones, but only when rare. However, blue males were not socially dominant when rare. The simulation results suggest that an own-morph attack bias reduces the social dominance of red males when they are more abundant. Thus, there is no evidence of symmetric negative frequency-dependent selection acting on social dominance, suggesting that additional fitness costs to the red morph must explain their co-existence.

No effect of blue on winning contests in judo.

A study by Rowe et al. reported a winning bias for judo athletes wearing a blue outfit relative to those wearing a white one during the 2004 Olympics. It was suggested that blue is associated with a higher likelihood of winning through differential effects of colour on opponent visibility and/or an intimidating effect on the opponent. However, we argue that there is no colour effect on winning in judo. We show that alternative factors, namely allocation biases, asymmetries in prior experience and differences in recovery time are possible confounding factors in the analysis of Rowe et al. After controlling for these factors, we found no difference in blue and white wins. We further analysed contest outcomes of 71 other major judo tournaments and also found no winning bias. Our findings have implications for sports policy makers: they suggest that a white-blue outfit pairing ensures an equal level of play.

Intrasexual competition among females and the stabilization of a conspicuous colour polymorphism in a Lake Victoria cichlid fish.

The maintenance of colour polymorphisms within populations has been a long-standing interest in evolutionary ecology. African cichlid fish contain some of the most striking known cases of this phenomenon. Intrasexual selection can be negative frequency dependent when males bias aggression towards phenotypically similar rivals, stabilizing male colour polymorphisms. We propose that where females are territorial and competitive, aggression biases in females may also promote coexistence of female morphs. We studied a polymorphic population of the cichlid fish Neochromis omnicaeruleus from Lake Victoria, in which three distinct female colour morphs coexist: one plain brown and two blotched morphs. Using simulated intruder choice tests in the laboratory, we show that wild-caught females of each morph bias aggression towards females of their own morph, suggesting that females of all three morphs may have an advantage when their morph is locally the least abundant. This mechanism may contribute to the establishment and stabilization of colour polymorphisms. Next, by crossing the morphs, we generated sisters belonging to different colour morphs. We find no sign of aggression bias in these sisters, making pleiotropy unlikely to explain the association between colour and aggression bias in wild fish, which is maintained in the face of gene flow. We conclude that female-female aggression may be one important force for stabilizing colour polymorphism in cichlid fish.