The contribution of executive functions to sex differences in animal cognition

Cognitive sex differences have been reported in several vertebrate species, mostly in spatial abilities. Here, I review evidence of sex differences in a family of general cognitive functions that control behaviour and cognition, i.e., executive functions such as cognitive flexibility and inhibitory control. Most of this evidence derives from studies in teleost fish. However, analysis of literature from other fields (e.g., biomedicine, genetic, ecology) concerning mammals and birds reveals that more than 40% of species investigated exhibit sex differences in executive functions. Among species, the direction and magnitude of these sex differences vary greatly, even within the same family, suggesting sex-specific selection due to species' reproductive systems and reproductive roles of males and females. Evidence also suggests that sex differences in executive functions might provide males and females highly differentiated cognitive phenotypes. To understand the evolution of cognitive sex differences in vertebrates, future research should consider executive functions

Evaluating guppy, Poecilia reticulata, as a model for sex differences in cognition.

Whether cognitive differences exist between men and women is a question probably as old as experimental psychology though still highly controversial. Despite the large number investigations, sex differences in performance emerge only in a few cases, such as in visuo-spatial or verbal abilities. However, it is not clear if these differences are biological or due to cultural influences, possibly because we know very little about their underlying mechanisms and evolutionary causes. Sex differences in cognition have been investigated also in few other species, such as monkeys, rodents and chickens. Rodents, in particular, represent a powerful model in the field of spatial abilities, allowing, among the others, complex hormonal manipulations, drugs tests and between-species comparative studies. Recent years have seen a massive increase in the use of fish as models for cognitive research, at the point that fish are predicted to replace classical vertebrates models soon. The existence of cognitive sex differences in fish has not been investigated. Paradoxically, experimenters quite often use indiscriminately fish of both sexes in their experiments, introducing a confound if cognitive sex differences exist. In this thesis, I studied cognitive sex differences in the guppy, Poecilia reticulata, one of the most studied species in behavioural and cognitive research. Male and female guppies are characterized by considerable differences in ecology and behaviour. I compared the performance of males and females in a wide range of cognitive tasks, including visual discrimination learning, novel object recognition, rule learning, reversal learning, spatial navigation, spatial learning and quantity discrimination. Male and female guppies showed similar abilities in solving most of the tasks, with only two exceptions. In a reversal learning task, guppies were initially trained to obtain food by choosing a predetermined colour between two options, a task that the two sexes learned equally well; then, the reward contingency was reversed and subjects had to inhibit the learned preference to select the other stimulus. Females quickly started to choose the new rewarded colour while males persisted longer in selecting the previously rewarded colour. The second difference emerged in a quantity discrimination task. Guppies were required to select the larger between two groups of conspecifics, following a natural tendency of social fish to stay in larger shoals to reduce predation risks. At the beginning of the experiment, females were much better than males at solving the task; this difference vanished after some minutes. The two observed sex differences are possibly the result of differential action of natural selection on the two sexes. Indeed, male guppies are thought to undergo selection for high persistence that helps in courting females intensively; females are likely to undergo strong selection for predator avoidance skills as predators preferentially target females. The absence of sex differences in the other cognitive abilities can be due to the absence of sex differences in selective pressures, or, alternatively, to the existence of constraints that prevent differentiation. In many of the tasks in which males and females obtained similar scores, they were nonetheless observed to behave quite differently. In visual discrimination tasks and in a spatial learning task, males were faster than females in deciding which option to choose, suggesting greater impulsivity in males. In the novel object recognition task, males and females expressed equal ability in discriminating the two objects, yet males explored the novel object at the beginning of the test, females some time afterwards. In the spatial navigation task, males persisted longer than females in trying to reach a target behind a transparent barrier. The general lack of sex differences in cognitive abilities in guppies aligns with the scarce evidence of sex differences in cognitive abilities in mammals. However, I provided evidence that minor differences in performance, such as in persistence, impulsivity and exploratory tendency, typically found in mammals and birds are widespread in fish as well

Familiarity effects on fish behaviour are disrupted in shoals that contain also unfamiliar individuals

Research on several social fishes has revealed that shoals constituted by familiar individuals behave remarkably differently compared to shoals formed by unfamiliar individuals. However, whether these behavioural changes may arise also in shoals composed by a mixture of familiar and unfamiliar individuals, a situation that may commonly occur in nature, is not clear. Here, we observed the behaviour of Mediterranean killifish (Aphanius fasciatus) shoals that were composed by both familiar and unfamiliar individuals (i.e. individuals were familiar to each other in pairs) and compared it with shoals entirely made by either unfamiliar or familiar individuals. Shoals formed by familiar individuals took longer to emerge from a refuge and swam more cohesively compared to shoals formed by unfamiliar fish. Shoals formed by a mixture of familiar and unfamiliar individuals behaved as shoals formed by unfamiliar individuals. Moreover, mixed shoals did not segregate in pairs according to their familiarity. This study suggests that mixed shoals do not show the behavioural effects of familiarity. Significance statement Laboratory studies have compared the behaviour of shoals formed by familiar fish versus shoals formed by unfamiliar fish, finding notable advantages in the former ones, such as improved antipredator and foraging behaviour. However, comparing these two opposite shoal types may not provide information on the natural situation, because in nature, shoals often change composition. We investigated how shoals formed by a mixture of familiar and unfamiliar fish behaved. We analysed shoals' preference for open environment versus covers and shoals' swimming cohesion. Results showed that shoals formed by both familiar and unfamiliar individuals mostly behave like shoals entirely formed by unfamiliar individuals. This suggests that the advantages of social groups formed by familiar fish might be hardly seen in nature for species in which shoal composition changes frequently

Interspecific differences in developmental mode determine early cognitive abilities in teleost fish

Most studies on developmental variation in cognition have suggested that individuals are born with reduced or absent cognitive abilities, and thereafter, cognitive performance increases with age during early development. However, these studies have been mainly performed in altricial species, such as humans, in which offspring are extremely immature at birth. In this work, we tested the hypothesis that species with other developmental modes might show different patterns of cognitive development. To this end, we analysed inhibitory control performance in two teleost species with different developmental modes, the zebrafish Danio rerio and the guppy Poecilia reticulata, exploiting a simple paradigm based on spontaneous behaviour and therefore applicable to subjects of different ages. Zebrafish hatch as larvae 3 days after fertilisation, and have an immature nervous system, a situation that mirrors extreme altriciality. We found that at the early stages of development, zebrafish displayed no evidence of inhibitory control, which only begun to emerge after one month of life. Conversely, guppies, which are born after approximately one month of gestation as fully developed and independent individuals, solved the inhibitory control task since their first days of life, although performance increased with sexual maturation. Our study suggests that the typical progression described during early ontogeny in humans and other species might not be the only developmental trend for animals' cognition and that a species' developmental mode might determine variation in cognition across subjects of different age

Daily rhythms in the behavioural stress response of the zebrafish Danio rerio

In nature, animals are exposed to stressors that occur with different likelihood throughout the day, such as risk of predation and human disturbance. Hence, the stress response is expected to vary plastically to adaptively match these challenges. Several studies have supported this hypothesis in a wide range of vertebrate species, including some teleost fish, mostly through evidence of circadian variation in physiology. However, in teleost fish, circadian variation in behavioural stress responses is less understood. Here, we investigated the daily rhythm of stress response at the behavioural level in the zebrafish Danio rerio. We exposed individuals and shoals to an open field test every 4 h over a 24 h cycle, recording three behavioural indicators of stress and anxiety levels in novel environments (thigmotaxis, activity and freezing). Thigmotaxis and activity significantly varied throughout the day with a similar pattern, in line with a stronger stress response in the night phase. The same was suggested by analysis of freezing in shoals, but not in individual fish, in which variation appeared mostly driven by a single peak in the light phase. In a control experiment, we observed a set of subjects after familiarisation with the open-field apparatus. This experiment indicated that activity and freezing might present a daily rhythmicity that is unrelated to environmental novelty, and thus to stress responses. However, the thigmotaxis was constant through the day in the control condition, suggesting that the daily variation of this indicator is mostly attributable to the stress response. Overall, this research indicates that behavioural stress response of zebrafish does follow a daily rhythm, although this may be masked using behavioural indicators other than thigmotaxis. This rhythmicity can be relevant to improve welfare in aquaculture and reliability of behavioural research in fish models

Quantity discrimination in newly hatched zebrafish suggests hardwired numerical abilities

An intriguing hypothesis to explain the ubiquity of numerical abilities is that all vertebrates are born with hardwired neuronal networks for processing numbers. To date, only studies on human foetuses have clearly supported this hypothesis. Zebrafish hatch 48–72 h after fertilisation with an embryonic nervous system, providing a unique opportunity for investigating this hypothesis. Here, we demonstrated that zebrafish larvae exposed to vertical bars at birth acquired an attraction for bar stimuli and we developed a numerical discrimination task based on this preference. When tested with a series of discriminations of increasing difficulty (1vs.4, 1vs.3, 1vs.2, and 2vs.4 bars), zebrafish larvae reliably selected the greater numerosity. The preference was significant when stimuli were matched for surface area, luminance, density, and convex hull, thereby suggesting a true capacity to process numerical information. Converging results from two phylogenetically distant species suggests that numerical abilities might be a hallmark feature of vertebrates’ brains

Comparison of anxiety-like and social behaviour in medaka and zebrafish

The medaka, Oryzias latipes, is rapidly growing in importance as a model in behavioural research. However, our knowledge of its behaviour is still incomplete. In this study, we analysed the performance of medaka in 3 tests for anxiety-like behaviour (open-field test, scototaxis test, and diving test) and in 3 sociability tests (shoaling test with live stimuli, octagonal mirror test, and a modified shoaling test with mirror stimulus). The behavioural response of medaka was qualitatively similar to that observed in other teleosts in the open-field test (thigmotaxis), and in 2 sociability tests, the shoaling test and in the octagonal mirror test (attraction towards the social stimulus). In the remaining tests, medaka did not show typical anxiety (i.e., avoidance of light environments and preference for swimming at the bottom of the aquarium) and social responses (attraction towards the social stimulus). As a reference, we compared the behaviour of the medaka to that of a teleost species with well-studied behaviour, the zebrafish, tested under the same conditions. This interspecies comparison indicates several quantitative and qualitative differences across all tests, providing further evidence that the medaka responds differently to the experimental settings compared to other fish models

Individual differences and knockout in zebrafish reveal similar cognitive effects of BDNF between teleosts and mammals

The remarkable similarities in cognitive performance between teleosts and mammals suggest that the underlying cognitive mechanisms might also be similar in these two groups. We tested this hypothesis by assessing the effects of the brain-derived neurotrophic factor (BDNF), which is critical for mammalian cognitive functioning, on fish's cognitive abilities. We found that individual differences in zebrafish's learning abilities were positively correlated with bdnf expression. Moreover, a CRISPR/Cas9 mutant zebrafish line that lacks the BDNF gene (bdnf(-/-)) showed remarkable learning deficits. Half of the mutants failed a colour discrimination task, whereas the remaining mutants learned the task slowly, taking three times longer than control bdnf(+/+) zebrafish. The mutants also took twice as long to acquire a T-maze task compared to control zebrafish and showed difficulties exerting inhibitory control. An analysis of habituation learning revealed that cognitive impairment in mutants emerges early during development, but could be rescued with a synthetic BDNF agonist. Overall, our study indicates that BDNF has a similar activational effect on cognitive performance in zebrafish and in mammals, supporting the idea that its function is conserved in vertebrates