Stiver KA, Harris RM, Townsend JP, Hofmann HA, Alonzo SH. Neural Gene Expression Profiles and Androgen Levels Underlie Alternative Reproductive Tactics in the Ocellated Wrasse, Symphodus ocellatus. Ethology. 2015;121 (2) :152 - 167.Abstract

Discrete variation in reproductive behavior and physiology is observed in diverse taxa. Although it is known that most within-sex alternative reproductive tactics arise as a consequence of phenotypic plasticity, relatively little is known about differential neural gene expression among plastic alternative reproductive phenotypes. In the ocellated wrasse Symphodus ocellatus, males exhibit one of three alternative tactics (nesting, satellite, and sneaker) within a reproductive season, but switch tactics between years. Satellites and sneakers spawn parasitically in dominant (nesting) males' nests, but only nesting males provide parental care. Nesting and satellite males show transient cooperative defense of nests against sneakers. Here, we analyze circulating sex steroid hormone levels and neural gene expression profiles in these three male phenotypes and in females. 11-ketotestosterone (but not testosterone) was highest in nesting males, while estradiol was highest in females. Brain transcriptomes of satellites and females were most similar to each other and intermediate to nesting and sneaker males. Sneakers showed more total expression differences, whereas nesting males showed higher magnitude expression differences. Our findings reveal the surprising extent to which neural gene expression patterns vary across reproductive tactics that vary in a number of social traits, including aggression, territoriality, and cooperation, providing important insights into the molecular mechanisms that may underlie variation in cooperative and reproductive behavior.

2015.stiver.ethology.pdf 2015.stiver.ethology.pdf
Harris RM, Hofmann HA. Seeing is believing: dynamic evolution of gene families. Proceedings of the National Academy of Sciences of the United States of America. 2015;112 (5) :1252 - 1253.Abstract

Educated by his deep appreciation of nature, Darwin observed that "from so simple a beginning endless forms most beautiful" have arisen throughout the evolutionary history of life on earth (1). The spectacular diversity of orchids (2) and beetles (3) has long fascinated naturalists and casual observers alike. More recently, the adaptive radiations of Hawaiian drosophilids (4), Caribbean Anolis lizards (5), and African cichlid fishes (6) have become prime examples for understanding the mechanisms that enable diversification. Gene duplication and deletion are generally considered important evolutionary mechanisms that give rise to phenotypic diversity (7). Following gene duplication and loss, adaptation and speciation appear to proceed through a combination of both structural and cis-regulatory changes in one or more paralogous genes (8). Recent advances in sequencing technology have enabled researchers to make significant progress in understanding the molecular evolution that has facilitated diversification. In PNAS, Cortesi et al. (9) examine the evolution of vertebrate opsin genes as a spectacular example of how gene duplication and deletion events that affect spectral sensitivity have driven adaptation to diverse light environments and visual displays.

Simões JM, Barata EN, Harris RM, O'Connell LA, Hofmann HA, Oliveira RF. Social odors conveying dominance and reproductive information induce rapid physiological and neuromolecular changes in a cichlid fish. BMC Genomics [Internet]. 2015;16 (1) :1 - 13. Publisher's VersionAbstract

Social plasticity is a pervasive feature of animal behavior. Animals adjust the expression of their social behavior to the daily changes in social life and to transitions between life-history stages, and this ability has an impact in their Darwinian fitness. This behavioral plasticity may be achieved either by rewiring or by biochemically switching nodes of the neural network underlying social behavior in response to perceived social information. Independent of the proximate mechanisms, at the neuromolecular level social plasticity relies on the regulation of gene expression, such that different neurogenomic states emerge in response to different social stimuli and the switches between states are orchestrated by signaling pathways that interface the social environment and the genotype. Here, we test this hypothesis by characterizing the changes in the brain profile of gene expression in response to social odors in the Mozambique Tilapia, Oreochromis mossambicus. This species has a rich repertoire of social behaviors during which both visual and chemical information are conveyed to conspecifics. Specifically, dominant males increase their urination frequency during agonist encounters and during courtship to convey chemical information reflecting their dominance status.

Taborsky M, Hofmann HA, Beery AK, Blumstein DT, Hayes LD, Lacey EA, Martins EP, Phelps SM, Solomon NG, Rubenstein DR. Taxon matters: Promoting integrative studies of social behavior: NESCent Working Group on Integrative Models of Vertebrate Sociality: Evolution, Mechanisms, and Emergent Properties. Trends in Neurosciences. 2015;38 (4) :189 - 191.Abstract

The neural and molecular mechanisms underlying social behavior - including their functional significance and evolution - can only be fully understood using data obtained under multiple social, environmental, and physiological conditions. Understanding the complexity of social behavior requires integration across levels of analysis in both laboratory and field settings. However, there is currently a disconnect between the systems studied in the laboratory versus the field. We argue that recent conceptual and technical advances provide exciting new opportunities to close this gap by making non-model organisms accessible to modern approaches in both laboratory and nature.

Yin W, Maguire SM, Pham B, Garcia AN, Dang NV, Liang J, Wolfe A, Hofmann HA, Gore AC. Testing the critical window hypothesis of timing and duration of estradiol treatment on hypothalamic gene networks in reproductively mature and aging female rats. Endocrinology. 2015;156 (8) :2918 - 2933.Abstract

At menopause, the dramatic loss of ovarian estradiol (E2) necessitates the adaptation of estrogen-sensitive neurons in the hypothalamus to an estrogen-depleted environment. We developed a rat model to test the "critical window" hypothesis of the effects of timing and duration of E2 treatment after deprivation on the hypothalamic neuronal gene network in the arcuate nucleus and the medial preoptic area. Rats at 2 ages (reproductively mature or aging) were ovariectomized and given E2 or vehicle replacement regimes of differing timing and duration. Using a 48-gene quantitative low-density PCR array and weighted gene coexpression network analysis, we identified gene modules differentially regulated by age, timing, and duration of E2 treatment. Of particular interest, E2 status differentially affected suites of genes in the hypothalamus involved in energy balance, circadian rhythms, and reproduction. In fact, E2 status was the dominant factor in determining gene modules and hormone levels; age, timing, and duration had more subtle effects. Our results highlight the plasticity of hypothalamic neuroendocrine systems during reproductive aging and its surprising ability to adapt to diverse E2 replacement regimes.

Harris RM, Hofmann HA. Neurogenomics of Behavioral Plasticity. Advances in Experimental Medicine and Biology [Internet]. 2014;781 :149-168. Publisher's VersionAbstract

Across animals, there is remarkable diversity in behavior. Modern genomic approaches have made it possible to identify the molecular underpinnings of varied behavioral phenotypes. By examining species with plastic phenotypes we have begun to understand the dynamic and flexible nature of neural transcriptomes and identified gene modules associated with variation in social and reproductive behaviors in diverse species. Importantly, it is becoming increasingly clear that some candidate genes and gene networks are involved in complex social behaviors across even divergent species, yet few comparative transcriptomics studies have been conducted that examine a specific behavior across species. We discuss the implications of a range of important and insightful studies that have increased our understanding of the neurogenomics of behavioral plasticity. Despite its successes, behavioral genomics has been criticized for its lack of hypotheses and causative insights. We propose here a novel avenue to overcome some of these short-comings by complementing "forward genomics" studies (i.e., from phenotype to behaviorally relevant gene modules) with a "reverse genomics" approach (i.e., manipulating novel gene modules to examine effects on behavior, hormones, and the genome itself) to examine the functional causes and consequences of differential gene expression patterns. We discuss how several established approaches (such as pharmacological manipulations of a novel candidate pathway, fine scale mapping of novel candidate gene expression in the brain, or identifying direct targets of a novel transcription factor of interest) can be used in combination with the analysis of the accompanying neurogenomic responses to reveal unexpected biological processes. The integration of forward and reverse genomics will move the field beyond statistical associations and yield great insights into the neural and molecular control of social behavior and its evolution.

Brewand D, Wagner CE, Li YI, Malinsky M, Keller I, Fan S, Simakov O, Ng AY, Lim ZW, Bezault E, et al. The genomic substrate for adaptive radiation in African cichlid fish. Nature [Internet]. 2014;513 :375-381. Publisher's VersionAbstract

Cichlid fishes are famous for large, diverse and replicated adaptive radiations in the Great Lakes of East Africa. To understand themolecularmechanisms underlying cichlid phenotypic diversity,wesequenced the genomesand transcriptomes of five lineages of Africancichlids: theNile tilapia (Oreochromis niloticus), an ancestral lineagewith low diversity; and four members of the East African lineage: Neolamprologus brichardi/pulcher (older radiation, Lake Tanganyika), Metriaclima zebra (recent radiation, LakeMalawi),Pundamilia nyererei (very recent radiation, LakeVictoria), andAstatotilapia burtoni (riverine species around Lake Tanganyika).Wefound an excess of gene duplications in the East African lineage compared to tilapia and other teleosts, an abundance of non-coding element divergence, accelerated coding sequence evolution, expression divergence associated with transposable element insertions, and regulation by novel microRNAs. In addition, we analysed sequence data from sixty individuals representing six closely related species from Lake Victoria, and show genome-wide diversifying selection on coding and regulatory variants, some of which were recruited fromancient polymorphisms. Weconclude that a numberof molecular mechanisms shaped East African cichlid genomes, and that amassing of standing variation during periods of relaxed purifying selectionmayhave been important in facilitating subsequent evolutionary diversification.

Harris RM, Dijkstra PD, Hofmann HA. Complex structural and regulatory evolution of the pro-opiomelanocortin gene family. General and Comparative Endocrinology [Internet]. 2014;195 :107 - 115. Publisher's VersionAbstract

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), ??-lipotropin hormone, and ??-endorphin. Here we examine the structure, tissue expression profile, and pattern of cis transcriptional regulation of the three pomc paralogs (??1, ??2, and ??) in the model cichlid fish Astatotilapia burtoni and other cichlids, teleosts, and mammals. We found that the hormone-encoding regions of pomc ??1, pomc ??2 and pomc ?? are highly conserved, with a few notable exceptions. Surprisingly, the pomc ?? gene of cichlids and pomacentrids (damselfish) encodes a novel melanocortin peptide, ??-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 ??1 and ??2 show a more spatially restricted expression profile than pomc ??. 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. ?? 2013 Elsevier Inc.

Smith CC, Harris RM, Lampert KP, Schartl M, Hofmann HA, Ryan MJ. Copy number variation in the melanocortin 4 receptor gene and alternative reproductive tactics the swordtail Xiphophorus multilineatus. Environmental Biology of Fishes. 2014;98 (1) :23 - 33. 2014.smith_.biofish.pdf
Hofmann HA, Beery AK, Blumstein DT, Couzin ID, Earley RL, Hayes LD, Hurd PL, Lacey EA, Phelps SM, Solomon NG, et al. An evolutionary framework for studying mechanisms of social behavior. Trends in Ecology and Evolution [Internet]. 2014;29 (10) :581 - 589. Publisher's VersionAbstract

Social interactions are central to most animals and have a fundamental impact upon the phenotype of an individual. Social behavior (social interactions among conspecifics) represents a central challenge to the integration of the functional and mechanistic bases of complex behavior. Traditionally, studies of proximate and ultimate elements of social behavior have been conducted by distinct groups of researchers, with little communication across perceived disciplinary boundaries. However, recent technological advances, coupled with increased recognition of the substantial variation in mechanisms underlying social interactions, should compel investigators from divergent disciplines to pursue more integrative analyses of social behavior. We propose an integrative conceptual framework intended to guide researchers towards a comprehensive understanding of the evolution and maintenance of mechanisms governing variation in sociality.

Weitekamp CA, Hofmann HA. Evolutionary themes in the neurobiology of social cognition. Current Opinion in Neurobiology [Internet]. 2014;28 :22 - 27. Publisher's VersionAbstract

Remarkable examples of social cognition have been described across a diverse range of species, yet surprisingly little is known about the neurobiological underpinnings of these behaviors. Recent studies suggest that the molecular pathways and neural networks that mediate social behavior have been relatively conserved across vertebrate evolution, suggesting that shared mechanisms may drive adaptive behavioral responses to social stimuli. Here, we review recent advances in the neurobiology of flexible and context-dependent social behaviors across vertebrate taxa, focusing on female mate choice, pair-bonding, and aggressive behavior. Furthermore, we highlight the outstanding opportunities for uncovering the mechanisms mediating cooperative behavior, an exemplar of social cognition. We suggest a framework for investigating context-dependent neural organization and the evoked neural response to social stimuli. ?? 2014 Elsevier Ltd.

Brawand D, Wagner CE, Li YI, Malinsky M, Keller I, Fan S, Simakov O, Ng AY, Lim ZW, Bezault E, et al. The genomic substrate for adaptive radiation in African cichlid fish. Nature. 2014;513 :375 - 381.Abstract

Cichlid fishes are famous for large, diverse and replicated adaptive radiations in the Great Lakes of East Africa. To under- standthemolecularmechanismsunderlying cichlidphenotypic diversity,wesequencedthegenomesandtranscriptomes of fivelineages of Africancichlids: theNile tilapia (Oreochromis niloticus),anancestral lineagewithlowdiversity;andfour members of the East African lineage: Neolamprologus brichardi/pulcher (older radiation, Lake Tanganyika),Metriaclima zebra(recent radiation,LakeMalawi),Pundamilianyererei (veryrecentradiation,LakeVictoria),andAstatotilapiaburtoni (riverine species aroundLakeTanganyika).Wefound an excess of gene duplications in the East African lineagecompared to tilapia and other teleosts, an abundance of non-coding element divergence, accelerated coding sequence evolution, expression divergence associated with transposable element insertions,and regulationby novel microRNAs. In addition, we analysed sequence data from sixty individuals representing six closely related species from Lake Victoria, and show genome-widediversifying selectiononcodingandregulatoryvariants,someofwhichwererecruited fromancientpoly- morphisms.Weconclude that anumberof molecular mechanismsshaped East African cichlid genomes, and that amass- ing of standing variation during periods of relaxed purifying selectionmayhavebeenimportantin facilitating subsequent evolutionary diversification.

Striedter GF, Belgard GT, Chen CC, Davis FP, Finlay BL, Güntürkün O, Hale ME, Harris JA, Hecht EE, Hof PR, et al. NSF workshop report: Discovering general principles of nervous system organization by comparing brain maps across species. Journal of Comparative Neurology. 2014;522 (7) :1445 - 1453.Abstract

Efforts to understand nervous system structure and function have received new impetus from the federal Brain Research through Advancing Innovative Neurotechnologies (BRAIN) Initiative. Comparative analyses can contribute to this effort by leading to the discovery of general principles of neural circuit design, information processing, and gene-structure-function relationships that are not apparent from studies on single species. We here propose to extend the comparative approach to nervous system 'maps' comprising molecular, anatomical, and physiological data. This research will identify which neural features are likely to generalize across species, and which are unlikely to be broadly conserved. It will also suggest causal relationships between genes, development, adult anatomy, physiology, and, ultimately, behavior. These causal hypotheses can then be tested experimentally. Finally, insights from comparative research can inspire and guide technological development. To promote this research agenda, we recommend that teams of investigators coalesce around specific research questions and select a set of 'reference species' to anchor their comparative analyses. These reference species should be chosen not just for practical advantages, but also with regard for their phylogenetic position, behavioral repertoire, well-annotated genome, or other strategic reasons. We envision that the nervous systems of these reference species will be mapped in more detail than those of other species. The collected data may range from the molecular to the behavioral, depending on the research question. To integrate across levels of analysis and across species, standards for data collection, annotation, archiving, and distribution must be developed and respected. To that end, it will help to form networks or consortia of researchers and centers for science, technology, and education that focus on organized data collection, distribution, and training. These activities could be supported, at least in part, through existing mechanisms at NSF, NIH, and other agencies. It will also be important to develop new integrated software and database systems for cross-species data analyses. Multidisciplinary efforts to develop such analytical tools should be supported financially. Finally, training opportunities should be created to stimulate multidisciplinary, integrative research into brain structure, function, and evolution.

Oldfield RG, Mandrekar K, Nieves XM, Hendrickson DA, Chakrabarty P, Swanson BO, Hofmann HA. Parental care in the Cuatro Cienegas cichlid, Herichthys minckleyi (Teleostei: Cichlidae). Hydrobiologia. 2014;748 (1) :233 - 257.Abstract

Behavioral studies have often examined parental care by measuring phenotypic plasticity of behavior within a species. Phylogenetic studies have compared parental care among species, but only at broad categories (e.g., care vs. no care). Here we provide a detailed account that integrates phylogenetic analysis with quantitative behavioral data to better understand parental care behavior in the Cuatro Ciénegas cichlid, Herichthys minckleyi. We found that H. minckleyi occurs in a clade of sexually monochromatic or weakly dichromatic monogamous species, but that male and female H. minckleyi have dramatically different reproductive coloration patterns, likely as a result of sexual selection. Furthermore, we found that males are polygynous; large males guard large territories, and smaller males may attempt alternative mating tactics (sneaking). Finally, compared to the closely related monogamous Rio Grande cichlid, H. cyanoguttatus, males of H. minckleyi were present at their nests less often and performed lower rates of aggressive offspring defense, and females compensated for the absence of their mates by performing higher levels of offspring defense. Body color, mating system, and parental care in H. minckleyi appear to have evolved after it colonized Cuatro Ciénegas, and are likely a result of evolution in an isolated, stable environment.

Fischer EK, Harris RM, Hofmann HA, Hoke KL. Predator exposure alters stress physiology in guppies across timescales. Hormones and Behavior [Internet]. 2014;65 (2) :165 - 172. Publisher's VersionAbstract

In vertebrates, glucocorticoids mediate a wide-range of responses to stressors. For this reason, they are implicated in adaptation to changes in predation pressure. Trinidadian guppies ( Poecilia reticulata) from high-predation environments have repeatedly and independently colonized and adapted to low-predation environments, resulting in parallel changes in life history, morphology, and behavior. We validated methods for non-invasive waterborne hormone sample collection in this species, and used this technique to examine genetic and environmental effects of predation on basal glucocorticoid (cortisol) levels. To examine genetic differences, we compared waterborne cortisol levels in high- and low-predation fish from two distinct population pairs. We found that fish from high-predation localities had lower cortisol levels than their low-predation counterparts. To isolate environmental influences, we compared waterborne cortisol levels in genetically similar fish reared with and without exposure to predator chemical cues. We found that fish reared with predator chemical cues had lower waterborne cortisol levels than those reared without. Comparisons of waterborne and whole-body cortisol levels demonstrated that populations differed in overall cortisol levels in the body, whereas rearing conditions altered the release of cortisol from the body into the water. Thus, evolutionary history with predators and lifetime exposure to predator cues were both associated with lower cortisol release, but depended on distinct physiological mechanisms. ?? 2013 Elsevier Inc.

Harris RM, Otopalik AG, Smith CS, Bucher D, Golowasch J, Hofmann HA. Single-Neuron gene expression analysis using the Maxwell ® 16 LEV System in the neural systems and behavior course. Promega. 2014;(January) :1 - 6. 2014.harris.promega.pdf
Oldfield RG, Harris RM, Hendrickson DA, Hofmann HA. Arginine vasotocin and androgen pathways are associated with mating system variation in North American cichlid fishes. Hormones and Behavior. 2013;64 (1) :44 - 52.Abstract

Neuroendocrine pathways that regulate social behavior are remarkably conserved across divergent taxa. The neuropeptides arginine vasotocin/vasopressin (AVT/AVP) and their receptor V1a mediate aggression, space use, and mating behavior in male vertebrates. The hormone prolactin (PRL) also regulates social behavior across species, most notably paternal behavior. Both hormone systems may be involved in the evolution of monogamous mating systems. We compared AVT, AVT receptor V1a2, PRL, and PRL receptor PRLR1 gene expression in the brains as well as circulating androgen concentrations of free-living reproductively active males of two closely related North American cichlid species, the monogamous Herichthys cyanoguttatus and the polygynous Herichthys minckleyi. We found that H. cyanoguttatus males bond with a single female and together they cooperatively defend a small territory in which they reproduce. In H. minckleyi, a small number of large males defend large territories in which they mate with several females. Levels of V1a2 mRNA were higher in the hypothalamus of H. minckleyi, and PRLR1 expression was higher in the hypothalamus and telencephalon of H. minckleyi. 11-ketotestosterone levels were higher in H. minckleyi, while testosterone levels were higher in H. cyanoguttatus. Our results indicate that a highly active AVT/V1a2 circuit(s) in the brain is associated with space use and social dominance and that pair bonding is mediated either by a different, less active AVT/V1a2 circuit or by another neuroendocrine system. ?? 2013 Elsevier Inc.

Huffman LS, O'Connell LA, Hofmann HA. Aromatase regulates aggression in the African cichlid fish Astatotilapia burtoni. Physiology and Behavior [Internet]. 2013;112-113 :77 - 83. Publisher's VersionAbstract

The roles of estrogen and androgens in male social behavior are well studied, but little is known about how these hormones contribute to behavior in a social hierarchy. Here we test the role of aromatase, the enzyme that converts testosterone into estradiol, in mediating aggression and reproductive behavior in male Astatotilapia burtoni, an African cichlid fish that displays remarkable plasticity in social behavior. We first measured aromatase expression in subordinate and dominant males in brain regions that regulate social behavior and found that subordinate males have higher aromatase expression than dominant males in the magnocellular and gigantocellular regions of the preoptic area. Next, we functionally tested the role of aromatase in regulating behavior by intraperitoneally injecting dominant males with either saline or fadrozole (FAD), an aromatase inhibitor, and found that FAD treatment decreases aggressive, but not reproductive, behaviors compared to saline controls. To determine the underlying physiological and molecular consequences of FAD treatment, we measured estradiol and testosterone levels from plasma and brain aromatase expression in FAD and saline treated dominant males. We found that estradiol levels decreased and testosterone levels increased in response to FAD treatment. Moreover, FAD treated males had increased aromatase expression in the gigantocellular portion of the POA, possibly a compensatory response. Overall, our results suggest aromatase is a key enzyme that promotes aggression in A. burtoni males through actions in the preoptic area. ?? 2013 Elsevier Inc.

Kidd MR, O'Connell LA, Kidd CE, Chen CW, Fontenot MR, Williams SJ, Hofmann HA. Female preference for males depends on reproductive physiology in the African cichlid fish Astatotilapia burtoni. General and Comparative Endocrinology [Internet]. 2013;180 (1) :56 - 63. Publisher's VersionAbstract

Mate choice is fundamental to sexual selection, yet little is known about underlying physiological mechanisms that influence female mating decisions. We investigated the endocrine underpinnings of female mate choice in the African cichlid Astatotilapia burtoni, a non-seasonal breeder. In addition to profiling behavioral and hormonal changes across the female reproductive cycle, we tested two hypotheses regarding possible factors influencing female mate choice. We first asked whether female mate choice is influenced by male visual and/or chemical cues. A. burtoni females were housed for one full reproductive cycle in the center of a dichotomous choice apparatus with a large (attractive) or small (unattractive) conspecific male on either side. Females associated mostly with small, less attractive males, but on the day of spawning reversed their preference to large, attractive males, with whom they mated almost exclusively, although this choice depended on the relative amount of androgens released into the water by small males. We next asked whether male behavior or androgen levels change in relation to the stimulus females' reproductive state. We found that stimulus male aggression decreased and reproductive displays increased as the day of spawning approached. Moreover male testosterone levels changed throughout the females' reproductive cycle, with larger males releasing more testosterone into the water than small males. Our data suggest that female association in a dichotomous choice assay is only indicative of the actual mate choice on the day of spawning. Furthermore, we show that male behavior and hormone levels are dependent on the reproductive state of conspecific females. ?? 2012 Elsevier Inc.

O'Connell LA, Fontenot MR, Hofmann HA. Neurochemical profiling of dopaminergic neurons in the forebrain of a cichlid fish, Astatotilapia burtoni. Journal of Chemical Neuroanatomy [Internet]. 2013;47 :106 - 115. Publisher's VersionAbstract

Across vertebrates, the mesolimbic reward system is a highly conserved neural network that serves to evaluate the salience of environmental stimuli, with dopamine as the neurotransmitter most relevant to its function. Although brain regions in the dopaminergic reward system have been well characterized in mammals, homologizing these brain areas with structures in teleosts has been controversial, especially for the mesencephalo-diencephalic dopaminergic cell populations. Here we examine the neurochemical profile of five dopaminergic cell groups (Vc, POA, PPr, TPp, pTn) in the model cichlid Astatotilapia burtoni to better understand putative homology relationships between teleosts and mammals. We characterized in the adult brain the expression patterns of three genes (etv5, nr4a2, and pitx3) that either specify dopaminergic cell fate or maintain dopaminergic cell populations. We then determined whether these genes are expressed in dopaminergic cells. We find many striking similarities in these gene expression profiles between dopaminergic cell populations in teleosts and their putative mammalian homologs. Our results suggest that many of these dopaminergic cell groups are indeed evolutionarily ancient and conserved across vertebrates. ?? 2013 Elsevier B.V.