Brain areas activated during visual learning in the cichlid fish Pseudotropheus zebra.

The neural correlates of most cognitive functions in fish are unknown. This project aimed to identify brain regions involved in visual learning in the cichlid fish Pseudotropheus zebra. The expression of the protein pS6 was measured in 19 brain areas and compared between groups of individuals subjected to four different behavioral contexts (control, avoidance, trained, and novelty groups). Control group individuals were sacrificed with minimal interactions. Fish in the avoidance group were chased with a net for an hour, after which they were sacrificed. Individuals in the trained group received daily training sessions to associate a visual object with a food reward. They were sacrificed the day they reached learning criterion. Fish in the novelty group were habituated to one set of visual stimuli, then faced a change in stimulus type (novelty stimulus) before they were sacrificed. Fish in the three treatment groups showed the largest activation of pS6 in the inferior lobes and the tectum opticum compared to the control group. The avoidance group showed additional activation in the preoptic area, several telencephalic regions, the torus semicircularis, and the reticular formation. The trained group that received a food reward, showed additional activation of the torus lateralis, a tertiary gustatory center. The only area that showed strong activation in all three treatment groups was the nucleus diffusus situated within the inferior lobe. The inferior lobe receives prominent visual input from the tectum via the nucleus glomerulosus but so far, nothing is known about the functional details of this pathway. Our study showed for the first time that the inferior lobes play an important role in visual learning and object recognition.

Cichlids and stingrays can add and subtract 'one' in the number space from one to five.

The numerical understanding of cichlids and stingrays was examined regarding addition and subtraction abilities within the number space of one to five. Experiments were conducted as two-alternative forced-choice experiments, using a delayed matching to sample technique. On each trial, fish had to perform either an addition or subtraction, based on the presentation of two-dimensional objects in two distinct colors, with the color signaling a particular arithmetic process. Six cichlids and four stingrays successfully completed training and recognized specific colors as symbols for addition and subtraction. Cichlids needed more sessions than stingrays to reach the learning criterion. Transfer tests showed that learning was independent of straightforward symbol memorization. Individuals did not just learn to pick the highest or lowest number presented based on the respective color; instead, learning was specific to adding or subtracting 'one'. Although group results were significant for both species in all tests, individual results varied. Addition was learned more easily than subtraction by both species. While cichlids learned faster than stingrays, and more cichlids than stingrays learned the task, individual performance of stingrays exceeded that of cichlids. Previous studies have provided ample evidence that fish have numerical abilities on par with those of other vertebrate and invertebrate species tested, a result that is further supported by the findings of the current study.

Neural substrates involved in the cognitive information processing in teleost fish.

Over the last few decades, it has been shown that fish, comprising the largest group of vertebrates and in many respects one of the least well studied, possess many cognitive abilities comparable to those of birds and mammals. Despite a plethora of behavioural studies assessing cognition abilities and an abundance of neuroanatomical studies, only few studies have aimed to or in fact identified the neural substrates involved in the processing of cognitive information. In this review, an overview of the currently available studies addressing the joint research topics of cognitive behaviour and neuroscience in teleosts (and elasmobranchs wherever possible) is provided, primarily focusing on two fundamentally different but complementary approaches, i.e. ablation studies and Immediate Early Gene (IEG) analyses. More recently, the latter technique has become one of the most promising methods to visualize neuronal populations activated in specific brain areas, both during a variety of cognitive as well as non-cognition-related tasks. While IEG studies may be more elegant and potentially easier to conduct, only lesion studies can help researchers find out what information animals can learn or recall prior to and following ablation of a particular brain area.

Seeing the forest before the trees-spatial orientation in freshwater stingrays (Potamotrygon motoro) in a hole-board task.

Freshwater stingrays (Potamotrygon motoro) have been shown to use a variety of spatial learning strategies including directional, landmark and place learning. In the present study, the significance of landmarks and geometric cues was investigated in a hole-board task. The aim was to determine cue preferences and collect additional information on the orientation mechanisms used in elasmobranchs. In four experiments, five juvenile stingrays had to memorize a fixed goal location within either a rectangular or a circular arena in the presence of goal-associated, signaling landmarks, proximal and distal cues. Transfer tests elucidated which cues the rays used or favored to reach the goal position. All rays successfully solved three of four tasks; as expected, different strategies were used in the process. Small alterations in the positioning of signaling landmarks (causing a spatial conflict between the previous feeding location and the new position of the signaling landmark) caused individuals to visit both locations equally often, whereas large alterations caused animals to ignore signaling cues and return to the previous feeding location. In the last and most complex experiment, three of five rays found the feeding location by remembering the positions of both proximate and distal landmarks in addition to memorizing particular swimming paths. Results showed that rays generally placed more importance on the overall environmental or geometric arrangement of the arena than on (individual) landmarks. This seems ecologically feasible, as distinct landmarks (e.g. rocks, pieces of wood, water plants) in the rays' natural environment may be more easily altered, removed or obscured from view than global ones (e.g. a river bend), which tend to be more stable. Overall, these results confirm those of previous studies, in that freshwater stingrays orient visually, learn quickly and can apply various orientation strategies, which are not mutually exclusive.

Perception and discrimination of movement and biological motion patterns in fish.

Vision is of primary importance for many fish species, as is the recognition of movement. With the exception of one study, assessing the influence of conspecific movement on shoaling behaviour, the perception of biological motion in fish had not been studied in a cognitive context. The aim of the present study was therefore to assess the discrimination abilities of two teleost species in regard to simple and complex movement patterns of dots and objects, including biological motion patterns using point and point-light displays (PDs and PLDs). In two-alternative forced-choice experiments, in which choosing the designated positive stimulus was food-reinforced, fish were first tested in their ability to distinguish the video of a stationary black dot on a light background from the video of a moving black dot presented at different frequencies and amplitudes. While all fish succeeded in learning the task, performance declined with decreases in either or both parameters. In subsequent tests, cichlids and damselfish distinguished successfully between the videos of two dots moving at different speeds and amplitudes, between two moving dot patterns (sinus vs. expiring sinus) and between animated videos of two moving organisms (trout vs. eel). Transfer tests following the training of the latter showed that fish were unable to identify the positive stimulus (trout) by means of its PD alone, thereby indicating that the ability of humans to spontaneously recognize an organism based on its biological motion may not be present in fish. All participating individuals successfully discriminated between two PDs and two PLDs after a short period of training, indicating that biological motions presented in form of PLDs are perceived and can be distinguished. Results were the same for the presentation of dark dots on a light background and light dots on a dark background.

Irrespective of size, scales, color or body shape, all fish are just fish: object categorization in the gray bamboo shark Chiloscyllium griseum.

Object categorization is an important cognitive adaptation, quickly providing an animal with relevant and potentially life-saving information. It can be defined as the process whereby objects that are not the same, are nonetheless grouped together according to some defining feature(s) and responded to as if they were the same. In this way, knowledge about one object, behavior or situation can be extrapolated onto another without much cost and effort. Many vertebrates including humans, monkeys, birds and teleosts have been shown to be able to categorize, with abilities varying between species and tasks. This study assessed object categorization skills in the gray bamboo shark Chiloscyllium griseum. Sharks learned to distinguish between the two categories, 'fish' versus 'snail' independently of image features and image type, i.e., black and white drawings, photographs, comics or negative images. Transfer tests indicated that sharks predominantly focused on and categorized the positive stimulus, while disregarding the negative stimulus.

Who would have thought that 'Jaws' also has brains? Cognitive functions in elasmobranchs.

Adaptation of brain structures, function and higher cognitive abilities most likely have contributed significantly to the evolutionary success of elasmobranchs, but these traits remain poorly studied when compared to other vertebrates, specifically mammals. While the pallium of non-mammalian vertebrates lacks the mammalian neocortical organization responsible for all cognitive abilities of mammals, several behavioural and neuroanatomical studies in recent years have clearly demonstrated that elasmobranchs, just like teleosts and other non-mammalian vertebrates, can nonetheless solve a multitude of cognitive tasks. Sharks and rays can learn and habituate, possess spatial memory; can orient according to different orientation strategies, remember spatial and discrimination tasks for extended periods of time, use tools; can imitate and learn from others, distinguish between conspecifics and heterospecifics, discriminate between either visual objects or electrical fields; can categorize visual objects and perceive illusory contours as well as bilateral symmetry. At least some neural correlates seem to be located in the telencephalon, with some pallial regions matching potentially homologous areas in other vertebrates where similar functions are being processed. Results of these studies indicate that the assessed cognitive abilities in elasmobranchs are as well developed as in teleosts or other vertebrates, aiding them in fundamental activities such as food retrieval, predator avoidance, mate choice and habitat selection.

No rainbow for grey bamboo sharks: evidence for the absence of colour vision in sharks from behavioural discrimination experiments.

Despite convincing data collected by microspectrophotometry and molecular biology, rendering sharks colourblind cone monochromats, the question of whether sharks can perceive colour had not been finally resolved in the absence of any behavioural experiments compensating for the confounding factor of brightness. The present study tested the ability of juvenile grey bamboo sharks to perceive colour in an experimental design based on a paradigm established by Karl von Frisch using colours in combination with grey distractor stimuli of equal brightness. Results showed that contrasts but no colours could be discriminated. Blue and yellow stimuli were not distinguished from a grey distractor stimulus of equal brightness but could be distinguished from distractor stimuli of varying brightness. In addition, different grey stimuli were distinguished significantly above chance level from one another. In conclusion, the behavioural results support the previously collected physiological data on bamboo sharks, which mutually show that the grey bamboo shark, like several marine mammals, is a cone monochromate and colourblind.

Symmetry perception in bamboo sharks (Chiloscyllium griseum) and Malawi cichlids (Pseudotropheus sp.).

Several species have been shown to perceive symmetry as a measure of superior genetic quality, useful for assessing potential mates or mediating other visual activities such as the selection of food sources. The current study assessed whether Pseudotropheus sp. and Chiloscyllium griseum, two fish species from distantly related groups, possess symmetry perception. In alternative two choice experiments, individuals were tested for spontaneous preferences and trained to discriminate between abstract symmetrical and asymmetrical stimulus pairs. Pair discriminations were followed by extensive categorization experiments. Transfer tests elucidated whether bilaterally symmetrical and rotationally symmetrical stimuli could be distinguished. Sharks were also tested for the degree of dissimilarity between two symbols that could still be detected. While sharks showed both a spontaneous preference for symmetry as well as remarkable discrimination abilities by succeeding in all of the presented tasks, cichlids showed no spontaneous preference, had difficulties in discriminating between symbols and performed poorly in the categorization experiments. Sharks distinguished between bilaterally and rotationally symmetrical stimuli and easily differentiated between a four-armed cross (all arms 90° apart) and a cross where one of the arms was only 45° spaced from the one next to it. Performance did not decline when the separation was extended to 70°, but was significantly reduced at an 80° separation. Results indicate that the ability for symmetry perception varies across fish species and individuals, whereby some can detect even subtle differences in this respect.

Visual discrimination of rotated 3D objects in Malawi cichlids (Pseudotropheus sp.): a first indication for form constancy in fishes.

Fish move in a three-dimensional environment in which it is important to discriminate between stimuli varying in colour, size, and shape. It is also advantageous to be able to recognize the same structures or individuals when presented from different angles, such as back to front or front to side. This study assessed visual discrimination abilities of rotated three-dimensional objects in eight individuals of Pseudotropheus sp. using various plastic animal models. All models were displayed in two choice experiments. After successful training, fish were presented in a range of transfer tests with objects rotated in the same plane and in space by 45° and 90° to the side or to the front. In one experiment, models were additionally rotated by 180°, i.e., shown back to front. Fish showed quick associative learning and with only one exception successfully solved and finished all experimental tasks. These results provide first evidence for form constancy in this species and in fish in general. Furthermore, Pseudotropheus seemed to be able to categorize stimuli; a range of turtle and frog models were recognized independently of colour and minor shape variations. Form constancy and categorization abilities may be important for behaviours such as foraging, recognition of predators, and conspecifics as well as for orienting within habitats or territories.

Visual discrimination and object categorization in the cichlid Pseudotropheus sp.

Object categorization is an important function of the visual system, quickly providing an animal with relevant information about its surrounding and current situation, as for example during predator detection. While the ability to categorize objects has already been observed in several vertebrate and even invertebrate species, no attempt has previously been made to evaluate this function in fish, the most species-rich vertebrate group. This study assessed form discrimination abilities and object categorization skills in the African cichlid Pseudotropheus sp. Fish could discriminate between a variety of two-dimensional geometrical shapes, forms and sizes and learned to distinguish between two categories, "fish" versus "snail". Performance remained undisturbed by extensive modifications to the stimuli, as long as key features were maintained. Results indicate that fish not only memorized the features of the positive stimulus (categorized the positive stimulus), but also categorized the negative stimulus. During transfer trials involving a previously unknown object, fish were able to discriminate between both the negative and the positive stimulus and the unknown stimulus and responded accordingly.