African Cichlid Fish Astatotilapia Burtoni Research Articles

Multiple behavioral mechanisms shape development in a highly social cichlid fish

Early-life social experiences shape adult phenotype, yet the underlying behavioral mechanisms remain poorly understood. We manipulated early-life social experience in the highly social African cichlid fish Astatotilapia burtoni to investigate the effects on behavior and stress axis function in juveniles. Juveniles experienced different numbers of social partners in stable pairs (1 partner), stable groups (6 fish; 5 partners), and socialized pairs (a novel fish was exchanged every 5 days; 5 partners). Treatments also differed in group size (groups vs. pairs) and stability (stable vs. socialized). We then measured individual behavior and water-borne cortisol to identify effects of early-life experience. We found treatment differences in behavior across all assays: open field exploration, social cue investigation, dominant behavior, and subordinate behavior. Treatment did not affect cortisol. Principal components (PC) analysis revealed robust co-variation of behavior across contexts, including with cortisol, to form behavioral syndromes sensitive to early-life social experience. PC1 (25.1 %) differed by social partner number: juveniles with more partners (groups and socialized pairs) were more exploratory during the social cue investigation, spent less time in the territory, and were more interactive as dominants. PC5 (8.5 %) differed by stability: socialized pairs were more dominant, spent less time in and around the territory, were more socially investigative, and had lower cortisol than stable groups or pairs. Observations of the home tanks provided insights into the social experiences that may underlie these effects. These results contribute to our understanding of how early-life social experiences are accrued and exert strong, lasting effects on phenotype.

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.

A behavioral logic underlying aggression in an African cichlid fish

AbstractSocial rank in a hierarchy determines which individuals have access to important resources such as food, shelter, and mates. In the African cichlid fish Astatotilapia burtoni, rank is under social control, such that larger males are more likely than smaller males to be dominant in rank. Although it is well known that the relative size of A. burtoni males is critical in controlling social rank, the specific behaviors underlying responses to males of different sizes are not well understood. In many other fish species, when size asymmetries between competing males are small, physical aggression is likely to be used to settle territorial disputes. Our goal was to characterize these responses in male A. burtoni by performing resident‐intruder assays, in which aggressive behaviors were measured in territorial males in response to the introduction of unfamiliar males that differed in relative standard length (SL). We found that the relative SL of intruders played an important role in determining behavioral performance. Resident males that were smaller than the intruder (resident >5% smaller in SL) or matched (resident between 0 and 5% smaller or larger in SL) in size to the intruder performed more lateral displays, a type of non‐physical aggression, than resident males that were larger than the intruder (resident >5% larger in SL). However, physical aggression, such as chases and bites, did not differ as a function of relative SL. Thus, contrary to what was expected based on work in other species, our results demonstrate that dominant resident A. burtoni increase non‐physical aggression to settle territorial disputes when non‐dominant intruders are closer in size. This behavioral strategy may aid in readily settling conflicts when size asymmetries are small.

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.

Radical change: temporal patterns of oxidative stress during social ascent in a dominance hierarchy

Dominant individuals have priority access to mates and resources. However, high rank can be costly too, especially when it is maintained by intense agonistic behavior. Oxidative stress has been proposed as a potential cost of social dominance. However, social dominance hierarchies can be dynamic, and few studies have examined the cost of social dominance when males are changing status. We studied temporal changes in markers of oxidative stress during social ascent in the East African cichlid fish Astatotilapia burtoni. After removing the dominant male, males ascended from subordinate to dominant status. On the first day of social ascent, immediately after the dominant male removal, the newly dominant male showed lower levels of plasma total antioxidant capacity (TAC). However, we found that liver TAC and liver superoxide dismutase (SOD), an enzymatic antioxidant, were significantly upregulated on day 1 and 2 of social ascent, respectively. No temporal variation in nicotinamide adenine dinucleotide phosphate-oxidase (NADPH-oxidase) activity, blood DNA damage, and liver DNA damage was observed. By day 14, all markers of oxidative stress were similar to those observed in stable dominant males, which had higher levels of reactive oxygen metabolites (ROM) compared to subordinate males. We conclude that markers of oxidative stress vary dramatically during social ascent in a time- and tissue-dependent manner. Our study provides a more nuanced look at the oxidative cost of social dominance and highlights the importance of considering temporal changes in markers of oxidative stress during important life-history events. High social status gives priority access to resources but also entails costs. Since dominant individuals tend to have higher metabolic rates, oxidative stress could be a potential cost of high social status. Oxidative stress can occur when reactive oxygen species, produced during metabolic activities, cannot be effectively neutralized by antioxidants, leading to oxidative cellular damage. Social hierarchies can be dynamic, yet how the cost of rank varies when males are changing social status is unknown. We allowed subordinate males to ascend to dominant status in the cichlid fish Astatotilapia burtoni. At the beginning of social ascent when males suddenly increase aggressive behavior, the total antioxidant defense was lower in blood plasma, as expected, when males suddenly increase their metabolism during aggressive interactions. However, antioxidant defense was increased in the liver suggesting that males can compensate for the potential cost of becoming dominant. Circulating reactive oxygen metabolites gradually increased over time reaching levels similar to stable dominant males, possibly due to males upregulating their reproductive axis. Our results highlight that markers of oxidative stress can vary dramatically over time during important events in life.

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.

Early-life social environment alters juvenile behavior and neuroendocrine function in a highly social cichlid fish

Early-life experiences can shape adult behavior, with consequences for fitness and health, yet fundamental questions remain unanswered about how early-life social experiences are translated into variation in brain and behavior. The African cichlid fish Astatotilapia burtoni, a model system in social neuroscience, is well known for its highly plastic social phenotypes in adulthood. Here, we rear juveniles in either social groups or pairs to investigate the effects of early-life social environments on behavior and neuroendocrine gene expression. We find that both juvenile behavior and neuroendocrine function are sensitive to early-life effects. Behavior robustly co-varies across multiple contexts (open field, social cue investigation, and dominance behavior assays) to form a behavioral syndrome, with pair-reared juveniles towards the end of syndrome that is less active and socially interactive. Pair-reared juveniles also submit more readily as subordinates. In a separate cohort, we measured whole brain expression of stress and sex hormone genes. Expression of glucocorticoid receptor 1a was elevated in group-reared juveniles, supporting a highly-conserved role for the stress axis mediating early-life effects. The effect of rearing environment on androgen receptor α and estrogen receptor α expression was mediated by treatment duration (1 vs. 5 weeks). Finally, expression of corticotropin-releasing factor and glucocorticoid receptor 2 decreased significantly over time. Rearing environment also caused striking differences in gene co-expression, such that expression was tightly integrated in pair-reared juveniles but not group-reared or isolates. Together, this research demonstrates the important developmental origins of behavioral phenotypes and identifies potential behavioral and neuroendocrine mechanisms.

Social context influences sensorimotor gating in female African cichlid fish Astatotilapia burtoni.

Disruption in prepulse inhibition (PPI), a sensorimotor gating phenomenon found in many species, has been associated with various psychiatric disorders in humans. Social defeat has been identified as a mediator of naturally evoked reductions of PPI in African cichlid fish Astatotilapia burtoni where males reversibly alter social status and their sensorimotor gating abilities. Here we investigated A. burtoni females, which establish a male-like social hierarchy with dominant (DOM) and subordinate (SUB) individuals when housed in communities without males. We asked if DOM and SUB females demonstrate socially induced PPI differences comparable to their male DOM and SUB counterparts. Results suggest that social defeat reduced PPI in SUB females as compared to DOM females (p = 0.033) and mixed-sex community female controls (p = 0.017). However, socially defeated females in same-sex communities remained proactive when engaging in antagonistic behaviors, which appears beneficial in avoiding substantial reductions in PPI as seen in reactive, socially defeated males. In open field swimming tests, SUB females exhibited increased anxiety-related behavior (thigmotaxis) as compared to females from mixed-sex communities (COM). Taken together, our results emphasize social defeat is a reliable modulator of PPI independent of sex, and anxiety related to social defeat might be a factor in mediating PPI plasticity.

Immigrant and extrinsic hybrid inviability contribute to reproductive isolation between lake and river cichlid ecotypes.

Understanding how reproductive barriers evolve and which barriers contribute to speciation requires the examination of organismal lineages that are still in the process of diversification and the study of the full range of reproductive barriers acting at different life stages. Lake and river ecotypes of the East African cichlid fish Astatotilapia burtoni show habitat-specific adaptations, despite different levels of genetic differentiation, and thus represent an ideal model to study the evolution of reproductive barriers. To evaluate the degree of reproductive isolation between genetically divergent lake and river populations, we performed a mesocosm mating experiment in a semi-natural setting at Lake Tanganyika. We assessed reproductive isolation in the presence of male-male competition by analyzing survival and growth rates of introduced adults and their reproductive success from genetic parentage of surviving offspring. The genetically divergent river population showed reduced fitness in terms of survival, growth rate, and mating success in a lake-like environment. Hybrid offspring between different populations showed intermediate survival consistent with extrinsic postzygotic reproductive barriers. Our results suggest that both prezygotic (immigrant inviability) and postzygotic reproductive barriers contribute to divergence, and highlight the value of assessing multiple reproductive barriers acting at different stages and in natural contexts to understand speciation mechanisms.

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.

Socially induced plasticity in sensorimotor gating in the African cichlid fish Astatotilapia burtoni.

Deficits in prepulse inhibition (PPI), social defeat and social withdrawal are hallmark features of several neurological and neuropsychiatric disorders. However, the link between social environment and PPI i.e., the possible role of social defeat in driving PPI plasticity, is far from clear. Here we explored these questions in the African cichlid fish Astatotilapia burtoni, where males exist as two distinct yet reversible phenotypes. In fish communities, DOMs exhibit frequent aggressive and territorial behaviors, threatening and attacking SUBs, which respond either by engaging in fights and fleeing, or by avoiding interaction with DOMs altogether. Social phenotypes were selected using focal observations of dominant and submissive behaviors. Tests of auditory PPI showed markedly decreased PPI in SUBs as compared to DOMs at prepulse/pulse interstimulus interval of ISI 50ms. Interestingly, further analysis showed the PPI reduction in SUBs was driven by males with low social interactivity. Testing males before and after social transitions revealed increasing and decreasing PPI in ascending and descending males, respectively. In an open field paradigm, SUBs also showed higher levels of wall hugging (thigmotaxis) and freezing when compared to DOMs i.e., an increase in anxiety-related behavior. Together the results suggest distinct yet reversible behavioral PPI phenotypes in A. burtoni males, and that social defeat drives PPI plasticity. The fact that PPI deficits are readily reversible by status change implies PPI plasticity may reflect an adaptive response to challenges in the social environment.

Neuromolecular Regulation of Aggression Differs by Social Role during Joint Territory Defense.

In response to a territory intrusion, neighboring males of the African cichlid fish Astatotilapia burtoni engage in aggressive joint territory defense in a manner that depends on their social role. Here, we examine the possible function of several neuroendocrine and neuromodulator pathways previously implicated in the regulation of complex social behavior. We find that the neuromolecular regulation of aggression during joint territory defense is very much dependent on an individual's role in this context. In neighbors but not in residents, aggression is correlated to gene expression in the medial part of the dorsal telencephalon (area Dm), the putative homolog to the mammalian basolateral amygdala. This correlation is strikingly high for expression of the serotonin receptor 5-HT2c, suggesting the serotonin system is important in regulating context-dependent behavior. Furthermore, by examining candidate gene expression co-variance patterns in area Dm and in the lateral part of the dorsal telencephalon (area Dl), the putative homolog to the mammalian hippocampus, we identify two main patterns: gene expression is co-regulated within, but not across, brain regions, and co-regulation is synergistic rather than antagonistic. Our results highlight the critical effect of social context on both behavior and its neuromolecular basis.

Reproductive and metabolic state differences in olfactory responses to amino acids in a mouth brooding African cichlid fish.

Olfaction mediates many crucial life-history behaviors such as prey detection, predator avoidance, migration and reproduction. Olfactory function can also be modulated by an animal's internal physiological and metabolic states. While this is relatively well studied in mammals, little is known about how internal state impacts olfaction in fishes, the largest and most diverse group of vertebrates. Here we apply electro-olfactograms (EOGs) in the African cichlid fish Astatotilapia burtoni to test the hypothesis that olfactory responses to food-related cues (i.e. l-amino acids; alanine and arginine) vary with metabolic, social and reproductive state. Dominant males (reproductively active, reduced feeding) had greater EOG magnitudes in response to amino acids at the same tested concentration than subordinate males (reproductively suppressed, greater feeding and growth rates). Mouth brooding females, which are in a period of starvation while they brood fry in their mouths, had greater EOG magnitudes in response to amino acids at the same tested concentration than both recovering and gravid females that are feeding. Discriminant function analysis on EOG magnitudes also grouped the male (subordinate) and female (recovering, gravid) phenotypes with higher food intake together and distinguished them from brooding females and dominant males. The slope of the initial negative phase of the EOG also showed intra-sexual differences in both sexes. Our results demonstrate that the relationship between olfaction and metabolic state observed in other taxa is conserved to fishes. For the first time, we provide evidence for intra-sexual plasticity in the olfactory response to amino acids that is influenced by fish reproductive, social and metabolic state.

Cognitive skills and the evolution of social systems.

How do animal social skills influence evolution? Complex animal social behaviors require many cognitive skills including individual recognition and observational learning. For social systems to evolve, these abilities need to be transmitted genetically or culturally and supported by the evolution of underlying neural systems. Because animal skill sets are so varied, it seems best to describe animal cognitive behaviors as being a social calculus that can change with experience, which has evolved to match and facilitate the complexity of the social system where it arose. That is, acquiring and using social information in response to a rapidly changing complex world leads to social competence enabling success in essential behavioral interactions. Here, we describe the remarkable suite of social skills discovered in the African cichlid fish Astatotilapia burtoni, including an attention hierarchy, male deception, transitive inference, the mechanistic bases of social dominance, female mate choice and the neural control of female reproductive behavior. The social calculus of this species is presented as an example of a potential causal factor in the evolution of sophisticated social behavior necessary for the evolutionary success of their social system.

Social plasticity in non-territorial male African cichlid fish Astatotilapia burtoni

The African cichlid fish Astatotilapia burtoni represents a valuable model system for studies of social decision-making due to its socially mediated phenotypic plasticity. The males reversibly transition in social status from reproductively dominant and territorial (DOM) to submissive and non-territorial (SUB). Males are traditionally categorized into these two behavioral phenotypes by observational scoring. There is evidence, however, that this dichotomous categorization might not capture the behavioral plasticity displayed by individuals transitioning between SUB and DOM status. To test this concept, we used focal observations of intrasexual conflict behavior in fish communities combined with a modified analysis of the ethogram typically used in A. burtoni. Results revealed a cluster of males close to the crossover point between SUB and DOM status as defined by the traditional dominance index. These intermediate males showed the highest frequency of intrasexual conflict behaviors, distinct behavioral responses to threats, and body pigment signaling displays that distinguish them from prototypical SUBs and DOMs. As such, our results provide a noninvasive behavioral metric to categorize A. burtoni males into three groups, thus further capturing the complex social dynamic of this model organism.

Localization of glutamatergic, GABAergic, and cholinergic neurons in the brain of the African cichlid fish, Astatotilapia burtoni.

Neural communication depends on release and reception of different neurotransmitters within complex circuits that ultimately mediate basic biological functions. We mapped the distribution of glutamatergic, GABAergic, and cholinergic neurons in the brain of the African cichlid fish Astatotilapia burtoni using in situ hybridization to label vesicular glutamate transporters (vglut1, vglut2.1, vglut3), glutamate decarboxylases (gad1, gad2), and choline acetyltransferase (chat). Cells expressing the glutamatergic markers vgluts 1-3 show primarily nonoverlapping distribution patterns, with the most widespread expression observed for vglut2.1, and more restricted expression of vglut1 and vglut3. vglut1 is prominent in granular layers of the cerebellum, habenula, preglomerular nuclei, and several other diencephalic, mesencephalic, and rhombencephalic regions. vglut2.1 is widely expressed in many nuclei from the olfactory bulbs to the hindbrain, while vglut3 is restricted to the hypothalamus and hindbrain. GABAergic cells show largely overlapping gad1 and gad2 expression in most brain regions. GABAergic expression dominates nuclei of the subpallial ventral telencephalon, while glutamatergic expression dominates nuclei of the pallial dorsal telencephalon. chat-expressing cells are prominent in motor cranial nerve nuclei, and some scattered cells lie in the preoptic area and ventral part of the ventral telencephalon. A localization summary of these markers within regions of the conserved social decision-making network reveals a predominance of either GABAergic or glutamatergic cells within individual nuclei. The neurotransmitter distributions described here in the brain of a single fish species provide an important resource for identification of brain nuclei in other fishes, as well as future comparative studies on circuit organization and function. J. Comp. Neurol. 525:610-638, 2017. © 2016 Wiley Periodicals, Inc.

Cobalt Chloride Treatment Used to Ablate the Lateral Line System Also Impairs the Olfactory System in Three Freshwater Fishes.

Fishes use multimodal signals during both inter- and intra-sexual displays to convey information about their sex, reproductive state, and social status. These complex behavioral displays can include visual, auditory, olfactory, tactile, and hydrodynamic signals, and the relative role of each sensory channel in these complex multi-sensory interactions is a common focus of neuroethology. The mechanosensory lateral line system of fishes detects near-body water movements and is implicated in a variety of behaviors including schooling, rheotaxis, social communication, and prey detection. Cobalt chloride is commonly used to chemically ablate lateral line neuromasts, thereby eliminating water-movement cues to test for mechanosensory-mediated behavioral functions. However, cobalt acts as a nonspecific calcium channel antagonist and could potentially disrupt function of all superficially located sensory receptor cells, including those for chemosensing. Here, we examined whether CoCl2 treatment used to ablate the lateral line system also impairs olfaction in three freshwater fishes, the African cichlid fish Astatotilapia burtoni, goldfish Carassius auratus, and the Mexican blind cavefish Astyanax mexicanus. To examine the impact of CoCl2 on the activity of peripheral receptors, we quantified DASPEI fluorescence intensity of the olfactory epithelium from fish exposed to control and CoCl2 solutions. In addition, we examined brain activation in olfactory processing regions of A. burtoni immersed in either control or cobalt solutions. All three species exposed to CoCl2 had decreased DASPEI staining of the olfactory epithelium, and in A. burtoni, cobalt treatment caused reduced neural activation in olfactory processing regions of the brain. To our knowledge this is the first empirical evidence demonstrating that the same CoCl2 treatment used to ablate the lateral line system also impairs olfactory function. These data have important implications for the use of CoCl2 in future research and suggest that previous studies using CoCl2 should be reinterpreted in the context of both impaired mechanoreception and olfaction.

Inhibition of Aromatase Induces Partial Sex Change in a Cichlid Fish: Distinct Functions for Sex Steroids in Brains and Gonads

Sex steroids are major drivers of sexual development and also responsible for the maintenance of the established gender. Especially fishes exhibit great plasticity and less conservation in sex determination and sexual development compared to other vertebrate groups. In addition, fishes have a constant sex steroid production throughout their entire lifespan, which makes them particularly susceptible to interferences with the endogenous sex steroid system. This susceptibility has recently been used to show that inhibition of the key enzyme of estrogen synthesis, aromatase Cyp19a1, can induce functional sex reversal even in adult fish. Here, we investigated the impact of the aromatase inhibitor (AI) fadrozole in adult females of the East African cichlid fish Astatotilapia burtoni. Using gene expression, phenotypic measurements, behavioral experiments, and hormone measurements, we assessed if females treated with fadrozole develop a male-like phenotype. We found that AI treatment has a different effect on gene expression in the gonad compared to the brain, the 2 tissues mostly implicated in sexual development. In contrast to observations in other gonochoristic species, A. burtoni ovaries cannot be transformed into functional testis by AI. However, rapid changes towards a male-like phenotype can be induced with AI in coloration, hormone levels, and behavior.

The mechanosensory lateral line is used to assess opponents and mediate aggressive behaviors during territorial interactions in an African cichlid fish

Fish must integrate information from multiple sensory systems to mediate adaptive behaviors. Visual, acoustic and chemosensory cues provide contextual information during social interactions, but the role of mechanosensory signals detected by the lateral line system during aggressive behaviors is unknown. The aim of this study was first to characterize the lateral line system of the African cichlid fish Astatotilapia burtoni and second to determine the role of mechanoreception during agonistic interactions. The A. burtoni lateral line system is similar to that of many other cichlid fishes, containing lines of superficial neuromasts on the head, trunk and caudal fin, and narrow canals. Astatotilapia burtoni males defend their territories from other males using aggressive behaviors that we classified as non-contact or contact. By chemically and physically ablating the lateral line system prior to forced territorial interactions, we showed that the lateral line system is necessary for mutual assessment of opponents and the use of non-contact fight behaviors. Our data suggest that the lateral line system facilitates the use of non-contact assessment and fight behaviors as a protective mechanism against physical damage. In addition to a role in prey detection, the diversity of lateral line morphology in cichlids may have also enabled the expansion of their social behavioral repertoire. To our knowledge, this is the first study to implicate the lateral line system as a mode of social communication necessary for assessment during agonistic interactions.