Spatial and non-spatial visual discrimination learning in zebrafish (Danio rerio)

Zebrafish (Danio rerio) provide an excellent model for assessment of molecular processes of neurodevelopment. To determine the functional importance of molecular events during neurodevelopment, we have developed methods for assessing learning in zebrafish in a three-chambered fish tank. In the first study, simple escape response was assessed. Zebrafish tested with a moving net learned to escape to another chamber more rapidly over the six sessions of training than the fish with the still net which did not learn. Upon reversal of the contingencies, the fish switched to the inactive net rapidly learned to suppress the escape response and fish formerly in the inactive net condition learned to avoid the moving net. In the second study, spatial discrimination learning was assessed. Zebrafish were trained on a right-left position discrimination to avoid the active net. Zebrafish showed significant improvement in escape responses over six sessions of training with three trials per session. In the third study, red-blue non-spatial discrimination learning was assessed. There was a significant improvement over the first six training sessions. With the reversal of contingencies, there was a significant decline of performance. With continued training, the fish again significantly improved avoidance. These studies found an effective motivational stimulus and procedure for studying escape behavior in zebrafish; a procedure whereby zebrafish would learn both spatial and non-spatial discrimination. These methods are being developed to help determine the functional importance of molecular events during zebrafish neurodevelopment. Accepted after revision: 20 August 2001 Electronic Publication     

Active avoidance conditioning in zebrafish (Danio rerio)

The zebrafish represents a potentially useful organism for studying genes involved in learning and memory function in vertebrates, because a number of genetic techniques in zebrafish have been developed to produce a wide variety of genetic mutants. While zebrafish mutants are being developed, behavioral studies on learning and memory function in zebrafish are in urgent need. The present study investigated active avoidance conditioning in normal zebrafish. Zebrafish were trained to swim from a lighted (CS) compartment to a dark compartment to avoid an electrical body shock (US) in a shuttle-box that consisted of a water-filled tank separated by an opaque barrier into two equal compartments. By varying the number of trials per training session and the duration of the intertrial interval, Experiments 1 and 2 showed that, with the CS, US, and intertrial interval being 12s, zebrafish learned avoidance responses within a training session consisting of 30 trials and retained the avoidance responses. Experiment 3 showed that zebrafish learned avoidance responses following the association between the CS of light and the US of shock in the avoidance conditioning paradigm. Using the avoidance conditioning paradigm, Experiment 4 investigated the amnestic effects of N-methyl-D-aspartate receptor antagonist MK-801 and nitric oxide synthase inhibitor L-NAME in zebrafish. Experiment 4 showed that post-training injection of L-NAME significantly impaired retention of avoidance responses while MK-801 did not, confirming previous results with other vertebrates. The results of the present study suggest the similar involvements of neurochemicals in learning and memory among vertebrates. Thus, future studies with zebrafish mutants may identify genes involved in learning and memory in vertebrates.     

Response acquisition by zebrafish (Danio rerio) with delayed reinforcement

Zebrafish (Danio rerio) is a common vertebrate animal model in biomedical research and is a promising species for studying how genes interact with environmental factors in determining behavior. The present study investigated how reinforcement parameters affect zebrafish behavior by assessing response acquisition with delayed reinforcement, which has been studied with other species (e.g., rats, pigeons, humans, etc.) but not with zebrafish. Twenty‐four experimentally naïve subjects were exposed to a tandem fixed‐ratio 1 differential‐reinforcement‐of‐other‐behavior x‐s schedule of reinforcement, where x varied across subjects. There were six different delay‐to‐reinforcement durations and sets of four fish were assigned to each delay duration. All of the fish assigned to a 0‐, 0.5‐, or 1‐s delay acquired responding. Two fish acquired responding with a 3‐s delay and one fish appeared to have acquired it with a 6‐s delay although the latter result was less clear. None acquired responding with a 12‐s delay. These results suggest that zebrafish behavior is sensitive to delays to reinforcement and the time frame over which reinforcement is effective may be limited approximately to 6 s. This time frame is shorter than that found with other species. Practical and theoretical implications of the present finding are discussed.     

Predator videos and electric shock function as punishers for zebrafish (Danio rerio)

Zebrafish (Danio rerio) are a promising animal model for studying the effects of gene–environment interactions on behavior. Two experiments were conducted to assess punishment effects of presenting predator videos (Indian leaf fish; Nandus nandus) and electric shock on operant approach responses in zebrafish. In Experiment 1, the predator video and shock stimuli were presented upon a response maintained by a single variable‐interval schedule of food reinforcement in different groups of fish. In Experiment 2, the predator video and shock stimuli were presented upon one of two response alternatives maintain by concurrently available variable‐interval schedules of food reinforcement in different groups of fish. Responding decreased when the predator video and shock stimuli were presented relative to their absence in both experiments. Moreover, responding on an unpunished alternative did not reliably decrease in Experiment 2. These results indicate that the decrease in responding resulted from the punishment contingency rather than from elicited species‐specific defense responses or conditioned avoidance. Thus, the predator video and electric shock functioned as punishers of operant behavior for zebrafish. Identifying punishers for this species could lead to research on how gene–environment interactions influence individual differences in sensitivity to punishment.     

Insensitivity to reward shifts in zebrafish (Danio rerio) and implications for assessing affective states

Animal Cognition - Theory and empirical findings predict that individuals in a negative affective state are more sensitive to unexpected reward loss and less sensitive to unexpected reward gain...     

Associative learning performance is impaired in zebrafish (Danio rerio) by the NMDA-R antagonist MK-801

The zebrafish is gaining popularity in behavioral neuroscience perhaps because of a promise of efficient large scale mutagenesis and drug screens that could identify a substantial number of yet undiscovered molecular players involved in complex traits. Learning and memory are complex functions of the brain and the analysis of their mechanisms may benefit from such large scale zebrafish screens. One bottleneck in this research is the paucity of appropriate behavioral screening paradigms, which may be due to the relatively uncharacterized nature of the behavior of this species. Here we show that zebrafish exhibit good learning performance in a task adapted from the mammalian literature, a plus maze in which zebrafish are required to associate a neutral visual stimulus with the presence of conspecifics, the rewarding unconditioned stimulus. Furthermore, we show that MK-801, a non-competitive NMDA-R antagonist, impairs memory performance in this maze when administered right after training or just before recall but not when given before training at a dose that does not impair motor function, perception or motivation. These results suggest that the plus maze associative learning paradigm has face and construct validity and that zebrafish may become an appropriate and translationally relevant study species for the analysis of the mechanisms of vertebrate, including mammalian, learning and memory.     

Strain differences in zebrafish (Danio rerio) social roles and their impact on group task performance

"Key" individuals in a social group are those that are centrally connected and thus serve as a hub for information flow across the group. From this position, they have the potential to have a powerful influence on group dynamics and performance. Here, we use metrics from social network theory to identify Key individuals in groups of 3-4 zebrafish (Danio rerio), and to measure the impact of removing those individuals from the group. We compared the results for two genetically distinct strains of zebrafish and found that although their social dynamics were superficially similar, one strain (Scientific Hatcheries, SH) responded to social perturbation, whereas the other (Parganas North, PN) did not. For both strains, groups that retained their Key fish performed better on a simple group foraging-learning task than did those from which the Key fish had been removed. However, the SH strain learned the task more quickly than did the PN strain, perhaps in part because of sex differences in task performance or because of strain differences in the reaction to experimental disturbance. We also confirm the utility of measures of social dynamics and social role that can be estimated reliably from very short observation sessions and by relatively untrained observers. These results set the stage for future research into the genetic mechanisms underlying social roles and group learning in vertebrates.     

Subcutaneous dye injection for marking and identification of individual adult zebrafish (Danio rerio) in behavioral studies

The zebrafish is increasingly utilized in behavioral brain research, as it offers a useful compromise between system complexity and practical simplicity. However, a potential drawback of this species in behavioral research is that individuals are difficult to distinguish. Here we describe a simple marking procedure, subcutaneous injection of color dyes, that may alleviate this problem. The procedure allowed us to successfully mark zebrafish and distinguish them for a period of more than 30 days, which is sufficiently long for most behavioral paradigms developed for this species. In addition, we also provide data suggesting that the injection-based marking does not significantly alter social interaction, as defined by the frequency of agonistic behaviors within shoals.     

Food quantity discrimination in puppies (Canis lupus familiaris)

Animal Cognition - There is considerable evidence that animals are able to discriminate between quantities. Despite the fact that quantitative skills have been extensively studied in adult...     

Sleep deprivation effects on object discrimination task in zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>)

The zebrafish is an ideal vertebrate model for neurobehavioral studies with translational relevance to humans. Many aspects of sleep have been studied, but we still do not understand how and why sleep deprivation alters behavioral and physiological processes. A number of hypotheses suggest its role in memory consolidation. In this respect, the aim of this study was to analyze the effects of sleep deprivation on memory in zebrafish (Danio rerio), using an object discrimination paradigm. Four treatments were tested: control, partial sleep deprivation, total sleep deprivation by light pulses, and total sleep deprivation by extended light. The control group explored the new object more than the known object, indicating clear discrimination. The partially sleep-deprived group explored the new object more than the other object in the discrimination phase, suggesting a certain degree of discriminative performance. By contrast, both total sleep deprivation groups equally explored all objects, regardless of their novelty. It seems that only one night of sleep deprivation is enough to affect discriminative response in zebrafish, indicating its negative impact on cognitive processes. We suggest that this study could be a useful screening tool for cognitive dysfunction and a better understanding of the effect of sleep-wake cycles on cognition.     

Large quantity discrimination by North Island robins (Petroica longipes)

While numerosity—representation and enumeration of different numbers of objects—and quantity discrimination in particular have been studied in a wide range of species, very little is known about the numerical abilities of animals in the wild. This study examined spontaneous relative quantity judgments (RQJs) by wild North Island robins (Petroica longipes) of New Zealand. In Experiment 1, robins were tested on a range of numerical values of up to 14 versus 16 items, which were sequentially presented and hidden. In Experiment 2, the same numerical contrasts were tested on a different group of subjects but quantities were presented as whole visible sets. Experiment 3 involved whole visible sets that comprised of exceedingly large quantities of up to 56 versus 64 items. While robins shared with other species a ratio-based representation system for representing very large values, they also appeared to have developed an object indexing system with an extended upper limit (well beyond 4) that may be an evolutionary response to ecological challenges faced by scatter-hoarding birds. These results suggest that cognitive mechanism influencing an understanding of physical quantity may be deployed more flexibly in some contexts than previously thought, and are discussed in light of findings across other mammalian and avian species.     

Shetland ponies (Equus caballus) show quantity discrimination in a matching-to-sample design

Numerical competence is one of the aspects of animal cognition with a long history of research interest, but few results are available for the horse. In the present study, we investigated the ability of three Shetland ponies to discriminate between different quantities of geometric symbols presented on a computer screen in a matching-to-sample arrangement. In Experiment 1, the ponies had to relate two similar quantities to another, paired in contrasts (1 vs. 2, 3 vs. 4 and 4 vs. 5) of the same stimulus (dot). Specific pairs of quantities (all differing by one) of up to five different geometrical symbols were displayed in Experiment 2. In each session, both quantities (more and less) were used as sample in such a way that each of the two quantities presented in one test served as positive and as negative stimulus, respectively. The three Shetland ponies were able to discriminate between the given quantities of dots by showing more than 80 % correct responses in two consecutive sessions. Only one of the ponies distinguished different shapes of geometric symbols at a level of 4 versus 5 items. The results show that all ponies were capable of visual quantity discrimination in the present matching-to-sample design, but task solving seemed more difficult when quantities were composed of heterogeneous stimuli. The present results confirm our hypothesis that the ponies based their decision on the matching concept of sameness and were not biased by a spontaneous preference for higher quantities.     

The discrimination of discrete and continuous amounts in African grey parrots (Psittacus erithacus)

A wealth of research in infants and animals demonstrates discrimination of quantities, in some cases nonverbal numerical perception, and even elementary calculation capacities. We investigated the ability of three African grey parrots (Psittacus erithacus) to select the largest amount of food between two sets, either discrete food items (experiment 1) or as volume of a food substance (experiment 2). The two amounts were presented simultaneously and were visible at the time of choice. Parrots were tested several times for all possible combinations between 1 and 5 seeds or 0.2 and 1 ml of food substance. In both conditions, subjects performed above chance for almost all combinations. Accuracy was negatively correlated with the ratio, that is performance improved with greater differences between amounts. Therefore, these results with both individual items and volume discrimination suggest that parrots use an analogue of magnitude, rather than object-file mechanisms to quantify items and substances.     

Quantity discrimination in fish species: fish use non-numerical continuous quantity traits to select shoals

Fish typically prefer to live in big shoals due to the associated ecological benefits. Shoaling is a behavior that depends on the ability to quantitatively discriminate. The fundamental mechanism involved in quantity discrimination determines whether fish can discriminate a shoal using numerical discrete cues (e.g., number of shoal members), non-numerical continuous traits (e.g., total body surface area) or both; however, the mechanism is currently a controversial topic. In the present study, we used a spontaneous choice experiment to test whether guppy (Poecilia reticulata), zebrafish (Danio rerio), Chinese crucian carp (Carassius auratus) and qingbo (Spinibarbus sinensis) rely on continuous (i.e., body surface area) or discrete (i.e., number of shoal members) information for shoal selection by altering the body surface area (cumulative body surface area ratio of 3:2 or 1:1) between two stimulus shoals with a different number of members (2 individuals vs 3 individuals). All four fish species preferred to shoal with the stimulus shoal with the larger cumulative surface area even if the shoal had fewer members; however, fish showed no shoal preference when the cumulative surface body areas of both stimulus shoals were equal. Furthermore, qingbo did not numerically discriminate between a shoal with 1 individual and a shoal with 3 individuals when the cumulative surface areas of both stimulus shoals were equal; however, qingbo showed a preference for the shoal with the larger cumulative surface area when the two stimulus shoals each had 3 individuals. In conclusion, the present study demonstrated that all four fish species relied only on non-numerical continuous quantity information for shoal selection, at least under a difficult task (i.e., 2 vs 3).     

Giraffes go for more: a quantity discrimination study in giraffes (Giraffa camelopardalis)

Animal Cognition - Many species, including humans, rely on an ability to differentiate between quantities to make decisions about social relationships, territories, and food. This study is the...     

Ratio-dependent quantity discrimination in quorum sensing ants

To optimise behaviour, organisms require information on the quantity of various components of their environment, and the ability of animals to discriminate quantity has been a subject of considerable recent interest. This body of research hints at generalised mechanisms of quantity discrimination in vertebrates, but data on invertebrates are still relatively scarce. In this study, I present data on the quantification abilities of an invertebrate in a novel context: quorum sensing. Quorum sensing generates a behavioural response in group-living animals once a threshold number of individuals, a ‘quorum’, is detected performing some key action. This process forms the basis for consensus decision-making in many species and allows group-living organisms to decide among mutually exclusive alternatives without compromising group integrity. To determine when a quorum is achieved, individuals must assess the number of group members performing the key action. Social insects employ quorum decisions to decide among potential nest sites when searching for a new home. In the Japanese ant, Myrmecina nipponica, quorum thresholds increase with colony size, providing an opportunity to assess the accuracy of quantity discrimination at different stimulus magnitudes. In this study, I demonstrate that the variation in individual quorum thresholds around the mean increases with increasing colony size. This indicates that the quantity discrimination ability of ants decreases with stimulus magnitude, and thus exhibits ratio dependence in the manner of Weber’s Law. This may have implications for the accuracy of consensus decision-making and other collective actions in a range of group-living organisms.     

The relative salience of discrete and continuous quantity in young infants

Whether human infants spontaneously represent number remains contentious. Clearfield & Mix (1999 ) and Feigenson, Carey & Spelke (2002 ) put forth evidence that when presented with small sets of 1–3 items infants may preferentially attend to continuous properties of stimuli rather than to number, and these results have been interpreted as evidence that infants may not have numerical competence. Here we present three experiments that test the hypothesis that infants prefer to represent continuous variables over number. In Experiment 1, we attempt to replicate the Clearfield & Mix study with a larger sample of infants. Although we replicated their finding that infants attend to changes in contour length, infants in our study attended to number and perimeter/area simultaneously. In Experiments 2 and 3, we pit number against continuous extent for exclusively large sets (Experiment 2) and for small and large sets combined (Experiment 3). In all three experiments, infants noticed the change in number, suggesting that representing discrete quantity is not a last resort for human infants. These results should temper the conclusion that infants find continuous properties more salient than number and instead suggest that number is spontaneously represented by young infants, even when other cues are available.     

Slowly walking down to the more food: relative quantity discrimination in African spurred tortoises (Centrochelys sulcata)

Animal Cognition - Quantity discrimination, is thought to be highly adaptive as it allows an organism to select greater amounts of food or larger social groups. In contrast to mammals, the...     

Performance of Asian elephants (Elephas maximus) on a quantity discrimination task is similar to that of African savanna elephants (Loxodonta africana)

Animal Cognition - Using an object-choice task, we measured the relative quantity discrimination ability of Asian elephants. Two zoo-housed elephants were given auditory cues of food being dropped...