Social ascent changes cognition, behaviour and physiology in a highly social cichlid fish

When an individual ascends in dominance status within their social community, they often undergo a suite of behavioural, physiological and neuromolecular changes. While these changes have been extensively characterized across a number of species, we know much less about the degree to which these changes in turn influence cognitive processes like associative learning, memory and spatial navigation. Here, we assessed male Astatotilapia burtoni , an African cichlid fish known for its dynamic social dominance hierarchies, in a set of cognitive tasks both before and after a community perturbation in which some individuals ascended in dominance status. We assayed steroid hormone (cortisol, testosterone) levels before and after the community experienced a social perturbation. We found that ascending males changed their physiology and novel object recognition preference during the perturbation, and they subsequently differed in social competence from non-ascenders. Additionally, using a principal component analysis we were able to identify specific cognitive and physiological attributes that appear to predispose certain individuals to ascend in social status once a perturbation occurs. These previously undiscovered relationships between social ascent and cognition further emphasize the broad influence of social dominance on animal decision-making. This article is part of the theme issue ‘The centennial of the pecking order: current state and future prospects for the study of dominance hierarchies’.

Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish

Direct tests of gene function have historically been performed in a limited number of model organisms. The CRISPR/Cas system is species-agnostic, offering the ability to manipulate genes in a range of models, enabling insights into evolution, development, and physiology. Astatotilapia burtoni, a cichlid fish from the rivers and shoreline around Lake Tanganyika, has been extensively studied in the laboratory to understand evolution and the neural control of behavior. Here we develop protocols for the creation of CRISPR-edited cichlids and create a broadly useful mutant line. By manipulating the Tyrosinase gene, which is necessary for eumelanin pigment production, we describe a fast and reliable approach to quantify and optimize gene editing efficiency. Tyrosinase mutants also remove a major obstruction to imaging, enabling visualization of subdermal structures and fluorophores in situ. These protocols will facilitate broad application of CRISPR/Cas9 to studies of cichlids as well as other non-traditional model aquatic species.

Spiny and soft-rayed fin domains in acanthomorph fish are established through a BMP-gremlin-shh signaling network

With over 18,000 species, the Acanthomorpha, or spiny-rayed fishes, form the largest and arguably most diverse radiation of vertebrates. One of the key novelties that contributed to their evolutionary success are the spiny rays in their fins that serve as a defense mechanism. We investigated the patterning mechanisms underlying the differentiation of median fin Anlagen into discrete spiny and soft-rayed domains during the ontogeny of the direct-developing cichlid fish Astatotilapia burtoni. Distinct transcription factor signatures characterize these two fin domains, whereby mutually exclusive expression of hoxa13a/b with alx4a/b and tbx2b marks the spine to soft-ray boundary. The soft-ray domain is established by BMP inhibition via gremlin1b, which synergizes in the posterior fin with shh secreted from a zone of polarizing activity. Modulation of BMP signaling by chemical inhibition or gremlin1b CRISPR/Cas9 knockout induces homeotic transformations of spines into soft rays and vice versa. The expression of spine and soft-ray genes in nonacanthomorph fins indicates that a combination of exaptation and posterior expansion of an ancestral developmental program for the anterior fin margin allowed the evolution of robustly individuated spiny and soft-rayed domains. We propose that a repeated exaptation of such pattern might underly the convergent evolution of anterior spiny-fin elements across fishes.

Control of testes mass by androgen receptor paralogs in a cichlid

Steroid hormones play numerous important and diverse roles in the differentiation and development of vertebrate primary and secondary reproductive characteristics. However, the exact role of androgen receptors (ARs)—which bind circulating androgens—in this regulatory pathway is unclear. Teleost fishes further complicate this question by having two paralogs of AR (ARα and ARβ) resulting from a duplication of their ancestral genome. We investigated the functional role of these two ARs on testes growth and development by experimentally eliminating receptor function of one or both paralogs using CRISPR/Cas9 genome edited Astatotilapia burtoni, an African cichlid fish. Fish with two or more functional receptor alleles were more likely to be male compared to fish with one or fewer, suggesting that the two paralogs of the receptor may be redundant in regulating early sex determination. In contrast, we found that adult testes size was significantly affected by distinct combinations of mutant and wild-type AR alleles. We present a working model whereby ARβ facilitates testes growth and ARα causes testes regression. This mechanism may contribute to the robust social and physiological plasticity displayed by A. burtoni and other social teleost fish.

Diversification dynamics and (non-)parallel evolution along an ecological gradient in African cichlid fishes

Understanding the drivers and dynamics of diversification is a central topic in evolutionary biology. Here, we investigated the dynamics of diversification in the cichlid fish Astatotilapia burtoni that diverged along a lake-stream environmental gradient. Whole-genome and morphometric analyses revealed that divergent selection was essential at the early stages of diversification, but that periods in allopatry were likely involved towards the completion of speciation. While morphological differentiation was continuous, genomic differentiation was not, as shown by two clearly separated categories of genomic differentiation. Reproductive isolation increased along a continuum of genomic divergence, with a “grey zone” of speciation at ~0.1% net nucleotide divergence. The quantification of the extent of (non-)parallelism in nine lake-stream population pairs from four cichlid species by means of multivariate analyses revealed one parallel axis of genomic and morphological differentiation among seven lake-stream systems. Finally, we found that parallelism was higher when ancestral lake populations were more similar.

Equal performance but distinct behaviors: Astatotilapia burtoni sex differences in a novel object recognition task and spatial maze

Sex differences in behavior and cognition can be driven by differential selection pressures from the environment and in the underlying neuromolecular mechanisms of decision-making. The highly social cichlid fish Astatotilapia burtoni exhibits dynamic and complex social hierarchies, yet explicit cognitive testing (outside of social contexts) and investigations of sex differences in cognition have yet to be fully explored. Here we assessed male and female A. burtoni in two cognitive tasks: a novel object recognition task and a spatial task. We hypothesized that males outperform females in a spatial learning task and exhibit more neophilic/exploratory behavior in across both tasks. In the present study we find that both sexes prefer the familiar object in a novel object recognition task, but the time at which they exhibit this preference differs between the sexes. Females more frequently learned the spatial task, exhibiting longer decision latencies and quicker error correction, suggesting a potential speed-accuracy tradeoff.Furthermore, the sexes differ in space use in both tasks and in a principal component analysis of the spatial task. A model selection analysis finds that preference, approach, and interaction duration in the novel object recognition task that reach a threshold of importance averaged across all models. This work highlights the need to explicitly test for sex differences in cognition to better understand how individuals navigate dynamic social environments.