Impacts of repeated social defeat on behavior and the brain in a cichlid fish.

Social defeat is a powerful experience leading to drastic changes in physiology and behavior, many of which are negative. For example, repeated social defeat in vertebrates results in reduced reproductive success, sickness and behavioral abnormalities that threaten individual survival and species persistence. However, little is known about what neural mechanisms are involved in determining whether an individual is resilient or susceptible to repeated social defeat stress. It also remains unknown whether exclusive use of reactive behaviors after repeated social defeat is maintained over time and impacts future behaviors during subsequent contests. We used a resident-intruder experiment in the African cichlid fish Astatotilapia burtoni to investigate the behavior and neural correlates of these two opposing groups. Behavior was quantified by watching fish during defeat trials and used to distinguish resilient and susceptible individuals. Both resilient and susceptible fish started with searching and freezing behaviors, with searching decreasing and freezing increasing after repeated social defeat. After a 4 day break period, resilient fish used both searching and freezing behaviors during a social defeat encounter with a new resident, while susceptible fish almost exclusively used freezing behaviors. By quantifying neural activation using pS6 in socially relevant brain regions, we identified differential neural activation patterns associated with resilient and susceptible fish and found nuclei that co-varied and may represent functional networks. These data provide the first evidence of specific conserved brain networks underlying social stress resilience and susceptibility in fishes.

Sensory Mechanisms of Parent-Offspring Recognition in Fishes, Amphibians, and Reptiles.

Parental care is important for offspring survival and success. Recognition of offspring by parents is critical to ensure parents direct care behaviors at related offspring and minimize energy lost by caring for unrelated young. Offspring recognition of parents prevents possible aggressive interactions between young and unrelated adults and allows offspring to direct begging behaviors toward the correct adult. Despite its importance and widespread nature, much of the current research has focused on a small range of species, particularly mammals and birds. We review the existing literature on the sensory mechanisms of parent-offspring recognition in fishes, amphibians, and reptiles. Within these groups, there is diversity in the presence and strategies for parent-offspring recognition. Future studies should continue to identify these mechanisms, as well as the neural and endocrine underpinnings in non-model organisms to expand our knowledge of this behavior and inform our understanding of the evolution of parent-offspring recognition.

Neural activation patterns associated with mouthbrooding, maternal care, infanticide and fry release in an African cichlid fish.

Parental care has evolved several times and is present across taxa. Parental care behaviors, such as food provisioning and protection, are critical for offspring success. However, infanticide can co-exist with parental care in the same species. The mechanisms underlying the switch from care to consumption and from offspring dependence to independence are relatively unknown, especially in fishes, the oldest and largest group of vertebrates. Mouthbrooding, an extreme example of parental care present in dozens of genera of fishes, provides an excellent opportunity to investigate the brain regions important for parental care. The maternal mouthbrooding African cichlid fish Astatotilapia burtoni broods developing young inside the mouth for approximately 14 days, then provides post-release maternal care by protecting fry inside the mouth when threatened. Following the post-release maternal care phase, females can exhibit infanticide and consume their own offspring. We used immunohistochemistry for the neural activation marker pS6 to identify differences in neural activation among mouthbrooding, maternal-care-providing and infanticide-exhibiting females, and between pre- and post-release fry. We identified five brain regions (Dc-5, ATn, nPPa, Vd-c and Dl-g) that are differentially activated among mouthbrooding, maternal care and infanticide females as well as six regions (Dm, Vv, Vd, Vs-m, TPp, PGZ and INL of retina) differentially activated between pre- and post-release fry. This study identifies both shared and distinct circuitry that may support transitions between parental care states and from care to infanticide, as well as regions in developed fry that support the transition from pre- to post-release.

Expression and localization of neuronal nitric oxide synthase in the brain and sensory tissues of the African cichlid fish Astatotilapia burtoni.

Nitric oxide (NO) produced by the enzyme neuronal nitric oxide synthase serves as an important neurotransmitter in the central nervous system that is involved in reproductive regulation, learning, sensory processing, and other forms of neural plasticity. Here, we map the distribution of nnos-expressing cells in the brain and retina of the cichlid fish Astatotilapia burtoni using in situ hybridization. In the brain, nnos-expressing cells are found from the olfactory bulbs to the hindbrain, including within specific nuclei involved in decision-making, sensory processing, neuroendocrine regulation, and the expression of social behaviors. In the retina, nnos-expressing cells are found in the inner nuclear layer, presumably in amacrine cells. We also used quantitative PCR to test for differences in nnos expression within the eye and olfactory bulbs of males and females of different reproductive states and social statuses. In the eye, males express more nnos than females, and socially dominant males express more nnos than subordinate males, but expression did not differ among female reproductive states. In the olfactory bulbs, dominant males had greater nnos expression than subordinate males. These results suggest a status-specific function for NO signaling in the visual and olfactory systems that may be important for sensory perception related to mating or territorial interactions to maintain the social hierarchy. The widespread distribution of nnos-expressing cells throughout the cichlid brain is similar to that in other teleosts, with some conserved localization patterns across vertebrates, suggesting diverse functions for this important neurotransmitter system.

Endocrine and neuroendocrine regulation of social status in cichlid fishes.

Position in a dominance hierarchy profoundly impacts group members' survival, health, and reproductive success. Thus, understanding the mechanisms that regulate or are associated with an individuals' social position is important. Across taxa, various endocrine and neuroendocrine signaling systems are implicated in the control of social rank. Cichlid fishes, with their often-limited resources of food, shelter, and mates that leads to competition, have provided important insights on the proximate and ultimate mechanisms related to establishment and maintenance of dominance hierarchies. Here we review the existing information on the relationships between endocrine (e.g., circulating hormones, gonadal and other tissue measures) and neuroendocrine (e.g., central neuropeptides, biogenic amines, steroids) systems and dominant and subordinate social rank in male cichlids. Much of the current literature is focused on only a few representative cichlids, particularly the African Astatotilapia burtoni, and several other African and Neotropical species. Many hormonal regulators show distinct differences at multiple biological levels between dominant and subordinate males, but generalizations are complicated by variations in experimental paradigms, methodological approaches, and in the reproductive and parental care strategies of the study species. Future studies that capitalize on the diversity of hierarchical structures among cichlids should provide insights towards better understanding the endocrine and neuroendocrine mechanisms contributing to social rank. Further, examination of this topic in cichlids will help reveal the selective pressures driving the evolution of endocrine-related phenotypic traits that may facilitate an individual's ability to acquire and maintain a specific social rank to improve survival and reproductive success.

Reproductive- and Social-State Plasticity of Multiple Sensory Systems in a Cichlid Fish.

Intra- and inter-sexual communications are vital to the survival and reproductive success of animals. In species that cycle in and out of breeding or other physiological condition, sensory function can be modulated to optimize communication at crucial times. Little is known, however, about how widespread this sensory plasticity is across taxa, whether it occurs in multiple senses or both sexes within a species, and what potential modulatory substances and substrates are involved. Thus, studying modulation of sensory communication in a single species can provide valuable insights for understanding how sensory abilities can be altered to optimize detection of salient signals in different sensory channels and social contexts. The African cichlid fish Astatotilapia burtoni uses multimodal communication in social contexts such as courtship, territoriality, and parental care and shows plasticity in sensory abilities. In this review, we synthesize what is known about how visual, acoustic, and chemosensory communication is used in A. burtoni in inter- and intra-specific social contexts, how sensory funtion is modulated by an individual's reproductive, metabolic, and social state, and discuss evidence for plasticity in potential modulators that may contribute to changes in sensory abilities and behaviors. Sensory plasticity in females is primarily associated with the natural reproductive cycle and functions to improve detection of courtship signals (visual, auditory, chemosensory, and likely mechanosensory) from high-quality males for reproduction. Plasticity in male sensory abilities seems to function in altering their ability to detect the status of other males in the service of territory ownership and future reproductive opportunities. Changes in different classes of potential modulators or their receptors (steroids, neuropeptides, and biogenic amines) occur at both peripheral sensory organs (eye, inner ear, and olfactory epithelium) and central visual, olfactory, and auditory processing regions, suggesting complex mechanisms contributing to plasticity of sensory function. This type of sensory plasticity revealed in males and females of A. burtoni is likely more widespread among diverse animals than currently realized, and future studies should take an integrative and comparative approach to better understand the proximate and ultimate mechanisms modulating communication abilities across taxa.

Opsin Expression Varies with Reproductive State in the Cichlid Fish Astatotilapia burtoni.

Animals use visual communication to convey crucial information about their identity, reproductive status, and sex. Plasticity in the auditory and olfactory systems has been well-documented, however, fewer studies have tested for plasticity in the visual system, a surprising detail since courtship and mate choice are largely dependent on visual signals across taxa. We previously found reproductive state-dependent plasticity in the eye of the highly social cichlid fish Astatotilapia burtoni. Male A. burtoni increase their courtship, including multicomponent visual displays, when around ovulated females, and ovulated females are more responsive to male visual courtship displays than non-ovulated females. Based on this, we hypothesized that ovulation status impacts visual capabilities in A. burtoni females. Using electroretinograms, we found that ovulated females had greater visual sensitivity at wavelengths corresponding to male courtship coloration compared with non-reproductively-receptive females. In addition, ovulated females had higher neural activation in the retina and higher mRNA expression levels of neuromodulatory receptors (e.g., sex-steroids; gonadotropins) in the eye than non-ovulated females. Here, we add to this body of work by testing the hypothesis that cone opsin expression changes with female reproductive state. Ovulated females had higher expression of short wavelength sensitive opsins (sws1, sws2a, sws2b) compared with mouthbrooding females. Further, expression of sws2a, the most abundant opsin in the A. burtoni eye, positively correlated with levels of circulating 11-ketotestosterone and estradiol and estrogen, androgen, and gonadotropin system receptor expression in the eye in females. These data indicate that reproductive state-dependent plasticity also occurs at the level of photoreceptors, not just through modulation of visual signals at downstream retinal layers. Collectively, these data provide crucial evidence linking endocrine modulation of visual plasticity to mate choice behaviors in females.

Expression patterns and evolution of urocortin and corticotropin-releasing hormone genes in a cichlid fish.

The corticotropin-releasing hormone and urocortin family of peptides consists of five members in many vertebrates: CRH (crha/crhb in teleosts), CRH2, UCN/UTS1, UCN2, and UCN3. These genes differ in expression pattern, as well as receptor affinity, allowing them to serve a wide range of functions in a variety of species. To better understand the roles of these genes in a single species, we examined their expression patterns in the cichlid fish Astatotilapia burtoni. In situ hybridization to map mRNA expression patterns of crhb, uts1, ucn2, and ucn3 in the brain revealed conserved and distinct spatial features of expression. crhb- and uts1-expressing cells were the most broadly distributed, with several areas of co-regionalization. ucn3 was less abundant but was found in discrete regions throughout the extent of the brain, with high expression in the cerebellum, while ucn2 was restricted to only a few areas. RT-PCR showed that while crhb, uts1, and ucn3 are found in several body tissues and widespread throughout the brain, ucn2 is quite restricted in the brain, and crha is only expressed in the eye. Bayesian phylogenetic analyses identified detailed relationships and novel orthologs in the urocortin family. We found evidence for a UCN2 gene loss in some reptiles. Our detailed description of the complete family of genes in the central nervous system of a model organism will inform future studies on the function of these genes in A. burtoni and provides a foundation for comparative studies with teleosts and other vertebrates.

Noise during mouthbrooding impairs maternal care behaviors and juvenile development and alters brain transcriptomes in the African cichlid fish Astatotilapia burtoni.

Anthropogenic noise has increased underwater ambient sound levels in the range in which most fishes detect and produce acoustic signals. Although the impacts of increased background noise on fish development have been studied in a variety of species, there is a paucity of information on how noise affects parental care. Mouthbrooding is an energetically costly form of parental care in which the brooding fish carries developing larvae in the buccal cavity for the duration of development. In the African cichlid Astatotilapia burtoni, females carry their brood for ~2 weeks during which time they do not eat. To test the hypothesis that increased background noise impacts maternal care behaviors and brood development, we exposed brooding females to a 3-h period of excess noise (~140 dB) played through an underwater speaker. Over half of noise-exposed brooding females cannibalized or pre-maturely released their brood, but 90% of control females exhibited normal brooding behaviors. RNA-seq analysis revealed that transcripts related to feeding and parental care were differentially expressed in the brains of noise-exposed females. Juveniles that were exposed to noise during their brood period within the mother's mouth had lower body condition factors, higher mortality and altered head transcriptomes compared with control broods. Furthermore, onset of adult-typical coloration and behaviors was delayed compared with control fish. Together, these data indicate that noise has severe impacts on reproductive fitness in mouthbrooding females. Our results, combined with past studies, indicate that parental care stages are extremely susceptible to noise-induced perturbations with detrimental effects on species persistence.

Female reproductive state is associated with changes in distinct arginine vasotocin cell types in the preoptic area of Astatotilapia burtoni.

Nonapeptides play a crucial role in mediating reproduction, aggression, and parental care across taxa. In fishes, arginine vasotocin (AVT) expression is related to social and/or reproductive status in most male fishes studied to date, and is linked to territorial defense, paternal care, and courtship. Despite a plethora of studies examining AVT in male fishes, relatively little is known about how AVT expression varies with female reproductive state or its role in female social behaviors. We used multiple methods for examining the AVT system in female African cichlid fish Astatotilapia burtoni, including immunohistochemistry for AVT, in situ hybridization for avt-mRNA, and quantitative PCR. Ovulated and mouthbrooding females had similar numbers of parvocellular, magnocellular, and gigantocellular AVT cells in the preoptic area. However, ovulated females had larger magnocellular and gigantocellular cells compared to mouthbrooding females, and gigantocellular AVT cell size correlated with the number of days brooding, such that late-stage brooding females had larger AVT cells than mid-stage brooding females. In addition, we found that ventral hypothalamic cells were more prominent in females compared to males, and were larger in mouthbrooding compared to ovulated females, suggesting a role in maternal care. Together, these data indicate that AVT neurons change across the reproductive cycle in female fishes, similar to that seen in males. These data on females complement studies in male A. burtoni, providing a comprehensive picture of the regulation and potential function of different AVT cell types in reproduction and social behaviors in both sexes.

Galanin neuron activation in feeding, parental care, and infanticide in a mouthbrooding African cichlid fish.

Galanin is a conserved neuropeptide involved in parental care and feeding. While galanin is known to mediate parental care and infanticide in rodents, its role in parental care and feeding behaviors in other taxa, particularly fishes, remains poorly understood. Mouthbrooding is an extreme form of parental care common in fishes in which caregivers carry offspring in their buccal cavity for the duration of development, resulting in obligatory starvation. In the cichlid fish Astatotilapia burtoni, females brood their young for ~2 wks and perform maternal care after release by collecting them into their mouth when threatened. However, females will cannibalize their brood after ~5 days. To examine the role of gal in feeding and maternal care, we collected mouthbrooding, fed, and starved females, as well as those displaying post-release maternal care and infanticide behaviors. Activation of gal neurons in the preoptic area (POA) was associated with parental care, providing the first link between gal and offspring-promoting behaviors in fishes. In contrast, activation of gal neurons in the lateral tuberal nucleus (NLT), the Arcuate homolog, was associated with feeding and infanticide. Overall, these data suggest gal is functionally conserved across vertebrate taxa with POA gal neurons promoting maternal care and Arc/NLT gal neurons promoting feeding.

Neural Activation Patterns Associated with Maternal Mouthbrooding and Energetic State in an African Cichlid Fish.

Parental care is widespread in the animal kingdom, but for many species, provisioning energetic resources must be balanced with trade-offs between self-promoting and offspring-promoting behaviors. However, little is known about the neural mechanisms underlying these motivational decisions. Mouthbrooding is an extreme form of parental care most common in fishes that provides an ideal opportunity to examine which brain regions are involved in parenting and energetics. The African cichlid fish Astatotilapia burtoni is a maternal mouthbrooder in which females hold developing young inside their mouths for 2 weeks. This brood care makes feeding impossible, so females undergo obligatory starvation. We used immunohistochemistry for the neural activation marker pS6 to examine which brain regions were involved in processing salient information in mouthbrooding, starved, and fed females. We identified brain regions more associated with maternal brood care (TPp, Dc-4/-5), and others reflective of energetic state (Dl-v, NLTi). Most nuclei examined, however, were involved in both maternal care and energetic status. Placement of each of the 16 examined nuclei into these functional categories was supported by node by node comparisons, co-activity networks, hierarchical clustering, and discriminant function analysis. These results reveal which brain regions are involved in parental care and food intake in a species where provisioning is skewed towards the offspring when parental feeding is not possible. This study provides support for both distinct and shared circuitry involved in regulation of maternal care, food intake, and energy balance, and helps put the extreme parental case of mouthbrooding into a comparative and evolutionary context.

Distribution of aromatase in the brain of the African cichlid fish Astatotilapia burtoni: Aromatase expression, but not estrogen receptors, varies with female reproductive-state.

Estrogen synthesis and signaling in the brains of vertebrates has pleotropic effects ranging from neurogenesis to modulation of behaviors. The majority of studies on brain-derived estrogens focus on males, but estrogenic signaling in females likely plays important roles in regulation of reproductive cycling and social behaviors. We used females of the mouth brooding African cichlid fish, Astatotilapia burtoni, to test for reproductive state-dependent changes in estrogenic signaling capacity within microdissected brain nuclei that are important for social behaviors. Expression levels of the rate-limiting enzyme aromatase, but not estrogen receptors, measured by qPCR changes across the reproductive cycle. Gravid females that are close to spawning had higher aromatase levels in all brain regions compared to females with lower reproductive potential. This brain aromatase expression was positively correlated with circulating estradiol levels and ovarian readiness. Using chromogenic in situ hybridization we localized aromatase-expressing cells to ependymal regions bordering the ventricles from the forebrain to the hindbrain, and observed more abundant staining in gravid compared to mouth brooding females in most regions. Staining was most prominent in subpallial telencephalic regions, and diencephalic regions of the preoptic area, thalamus, and hypothalamus, but was also observed in sensory and sensorimotor areas of the midbrain and hindbrain. Aromatase expression was observed in radial glial cells, revealed by co-localization with the glial marker GFAP and absence of co-localization with the neuronal marker HuC/D. Collectively these results support the idea that brain-derived estradiol in females may serve important functions in reproductive state-dependent physiological and behavioral processes across vertebrates.

Reproductive state-dependent plasticity in the visual system of an African cichlid fish.

Visual communication is used widely across the animal kingdom to convey crucial information about an animals' identity, reproductive status, and sex. Although it is well-demonstrated that auditory and olfactory sensitivity can change with reproductive state, fewer studies have tested for plasticity in the visual system, a surprising detail since courtship and mate choice behaviors in many species are largely dependent on visual signals. Here, we tested for reproductive state-dependent plasticity in the eye of the cichlid fish Astatotilapia burtoni using behavioral, gene expression, neural activation, and electrophysiology techniques. Males court ovulated females more intensely than gravid females, and ovulated females were more responsive to male courtship behaviors than gravid females. Using electroretinography to measure visual sensitivity in dark-adapted fish, we revealed that gravid, reproductively-ready females have increased visual sensitivity at wavelengths associated with male courtship coloration compared to non-gravid females. After ovulation was hormonally induced, female's spectral sensitivity further increased compared to pre-injection measurements. This increased sensitivity after hormone injection was absent in non-gravid females and in males, suggesting an ovulation-triggered increase in visual sensitivity. Ovulated females had higher mRNA expression levels of reproductive neuromodulatory receptors (sex-steroids; gonadotropins) in the eye than nonovulated females, whereas males had similar expression levels independent of reproductive/social state. In addition, female mate choice-like behaviors positively correlated with expression of gonadotropin system receptors in the eye. Collectively, these data provide crucial evidence linking endocrine modulation of visual plasticity to mate choice behaviors in females.

Mechanistic target of rapamycin (mTOR) implicated in plasticity of the reproductive axis during social status transitions.

The highly conserved brain-pituitary-gonadal (BPG) axis controls reproduction in all vertebrates, so analyzing the regulation of this signaling cascade is important for understanding reproductive competence. The protein kinase mechanistic target of rapamycin (mTOR) functions as a conserved regulator of cellular growth and metabolism in all eukaryotes, and also regulates the reproductive axis in mammals. However, whether mTOR might also regulate the BPG axis in non-mammalian vertebrates remains unexplored. We used complementary experimental approaches in an African cichlid fish, Astatotilapia burtoni, to demonstrate that mTOR is involved in regulation of the brain, pituitary, and testes when males rise in rank to social dominance. mTOR or downstream components of its signaling pathway (p-p70S6K) were detected in gonadotropin-releasing hormone (GnRH1) neurons, the pituitary, and testes. Transcript levels of mtor in the pituitary and testes also varied when reproductively-suppressed subordinate males rose in social rank to become dominant reproductively-active males, a transition similar to puberty in mammals. Intracerebroventricular injection of the mTORC1 inhibitor, rapamycin, revealed a role for mTOR in the socially-induced hypertrophy of GnRH1 neurons. Rapamycin treatment also had effects at the pituitary and testes, suggesting involvement of the mTORC1 complex at multiple levels of the reproductive axis. Thus, we show that mTOR regulation of BPG function is conserved to fishes, likely playing important roles in regulating reproduction and fertility across all male vertebrates.

Male dominance status regulates odor-evoked processing in the forebrain of a cichlid fish.

The ability to identify odors in the environment is crucial for survival and reproduction. However, whether olfactory processing in higher-order brain centers is influenced by an animal's physiological condition is unknown. We used in vivo neuron and local field potential (LFP) recordings from the ventral telencephalon of dominant and subordinate male cichlids to test the hypothesis that response properties of olfactory neurons differ with social status. Dominant males had a high percentage of neurons that responded to several odor types, suggesting broad tuning or differential sensitivity when males are reproductively active and defending a territory. A greater percentage of neurons in dominant males also responded to sex- and food-related odors, while a greater percentage of neurons in subordinate males responded to complex odors collected from behaving dominant males, possibly as a mechanism to mediate social suppression and allow subordinates to identify opportunities to rise in rank. Odor-evoked LFP spectral densities, indicative of synaptic inputs, were also 2-3-fold greater in dominant males, demonstrating status-dependent differences in processing possibly linking olfactory and other neural inputs to goal-directed behaviors. For the first time we reveal social and reproductive-state plasticity in olfactory processing neurons in the vertebrate forebrain that are associated with status-specific lifestyles.

Expression of tachykinin3 and related reproductive markers in the brain of the African cichlid fish Astatotilapia burtoni.

Neurokinin B, encoded by the tachykinin3 gene, plays a crucial role in regulating reproduction in mammals via KNDy neurons and interaction with GnRH. Previous work in teleost fishes has focused on hypothalamic tac3 expression for its role in reproduction, but detailed studies on extra-hypothalamic tac3 expression are limited. Here, we identified two tac3 genes in the social African cichlid fish Astatotilapia burtoni, only one of which produces a functional protein containing the signature tachykinin motif. In situ hybridization for tac3a mRNA identified cell populations throughout the brain. Numerous tac3a cells lie in several thalamic and hypothalamic nuclei, including periventricular nucleus of posterior tuberculum, lateral tuberal nucleus (NLT), and nucleus of the lateral recess (NRL). Scattered tac3-expressing cells are also present in telencephalic parts, such as ventral (Vv) and supracomissural (Vs) part of ventral telencephalon. In contrast to other teleosts, tac3 expression was absent from the pituitary. Using double-fluorescent staining, we localized tac3a-expressing cells in relation to GnRH and kisspeptin cells. Although no GnRH-tac3a colabeled cells were observed, dense GnRH fibers surround and potentially synapse with tac3a cells in the preoptic area. Only minimal (<5%) colabeling of tac3a was observed in kiss2 cells. Despite tac3a expression in many nodes of the mesolimbic reward system, it was absent from tyrosine hydroxylase (TH)-expressing cells, but tac3a cells were located in areas with dense TH fibers. The presence of tac3a-expressing cells throughout the brain, including in socially relevant brain regions, suggest more diverse functions beyond regulation of reproductive physiology that may be conserved across vertebrates.

Sexually-Relevant Visual and Chemosensory Signals Induce Distinct Behaviors and Neural Activation Patterns in the Social African Cichlid, Astatotilapia burtoni.

Across vertebrates, the use of multimodal (multiple sensory modalities) signals has evolved to convey important information to receivers. Information content of multimodal signals can be the same as or different from information in each unimodal signal, and are classified as redundant or non-redundant, respectively, based on receivers' behavioral responses. Despite the prevalence and importance of multimodal signaling across taxa, relatively little is known about how and where these signals are processed in the brains of receivers. We used the social African cichlid fish, Astatotilapia burtoni, to investigate how sexually-relevant visual and chemosensory uni- and multimodal signals from gravid (full of eggs) females influence behavior, brain activation patterns, and physiology in dominant males. We presented both visual and chemosensory signals either alone or together and found that males need sexually-relevant visual signals to engage in stereotypical courtship behaviors such as body quivers, waggles, and leads into spawning territories. However, the number of courtship behaviors was greater when males were exposed to multimodal visual-chemosensory signals, compared to either unimodal signal alone. When a female visual signal was absent, males increased swimming and overall activity in response to female-conditioned water compared to control water, suggesting that female-released chemosensory signals may stimulate male searching behavior and motivation. Importantly, we also tested anosmic (olfactory ablated) males to demonstrate that this behavior is primarily mediated by the olfactory system rather than gustation. Using the immediate early gene cfos as a proxy for neural activation, we also demonstrate differential activation in social and olfactory-relevant brain regions of dominant males exposed to unimodal and multimodal visual-chemosensory signals. We found at least one region that is preferentially activated by reception of signals from each sense, as well as regions that exhibit an additive effect on activation with multimodal visual-chemosensory stimulation. These data provide insight on how multimodal signals are processed in the brain and integrated with internal physiology of receivers to produce social behaviors, and lay the groundwork for future studies on the evolution of sensory perception.

Neural and behavioural correlates of repeated social defeat.

Dominance hierarchies are common across the animal kingdom and have important consequences for reproduction and survival. Animals of lower social status cope with repeated social defeat using proactive and reactive behaviours. However, there remains a paucity of information on how an individual's coping behaviours changes over time or what neural mechanisms are involved. We used a resident-intruder paradigm in the African cichlid fish Astatotilapia burtoni to investigate the neural correlates of these two opposing behaviour groups. Fish initially used both proactive and reactive behaviours, but had a dramatic increase in use of proactive behaviours during the third interaction, and this was followed by cessation of proactive behaviours and exclusive use of reactive coping. By quantifying neural activation in socially-relevant brain regions, we identify a subset of brain nuclei, including those homologous to the mammalian amygdala, showing higher activation in fish displaying proactive but not reactive behaviours. Fish displaying reactive behaviours had greater neural activation in the superior raphe, suggesting a possible conserved function during social defeat across vertebrates. These data provide the first evidence on the involvement of specific brain regions underlying proactive and reactive coping in fishes, indicating that these nuclei have conserved functions during social defeat across taxa.

Astatotilapia burtoni: A Model System for Analyzing the Neurobiology of Behavior.

Most biomedical research is performed using a very limited number of "model" species. In part, this has resulted from a combination of full genomes, manipulation of genes, and short generation times in these species. However, the advent of low-cost sequencing and gene editing in any organism has increased the use of nontraditional organisms. Many scientists have paraphrased the adage by Krogh [ Krogh , A. ( 2018 ) Science 70 , 200 - 204 ] that for many biological problems some species will prove to be most convenient and useful to study. In particular, using organisms most suited to the specific research question can lead to novel insights about fundamental physiological, neurobiological, immunological, and neuroendocrine systems that can advance our understanding of the well-being and health of humans. In addition, such studies have led to new ideas about the evolution and mechanisms that control social behavior. Fishes constitute about 50% of all vertebrate species and are the most diverse vertebrate radiation. Here we review behavioral and neurobiological discoveries of plasticity in social behavior resulting from analysis of an African cichlid fish, showing how its unique behavioral system has facilitated a broad range of discoveries. For many future questions, Astatotilapia burtoni and other cichlid fishes may be ideally suited to study as we advance our understanding of the neural basis of social decisions.