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The Zebrafish Y Maze was initially validated in a study by Cognato et al. (2012). The Y-Maze memory task involves a straightforward and quick training session focused on novelty exploration. Zebrafish tend to spend more time in the novel arm of the Y-Maze compared to the other arms, demonstrating responses to both novelty and spatial memory during training and testing intervals.
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Zebrafish Y-Maze: An Aquatic Behavioral Assessment Tool
The Zebrafish Y-Maze represents an aquatic adaptation of the traditional Y-Maze utilized in rodent studies. This maze, a modified version of the T-Maze, features a more natural angle of the arms (120 degrees compared to 90 degrees in the T-Maze), making it a preferred choice for behavioral assessments. Historically, the Y-Maze has been instrumental in evaluating learning and memory functions in rodent models (Yamazaki et al. 1979; Conrad et al. 1996; Van der Borght et al. 2007).
Rising Popularity of Zebrafish as a Model Organism
Zebrafish has steadily gained prominence as a model organism. This aquatic vertebrate exhibits significant physiological and genetic similarity to humans and is easily amenable to genetic manipulations for diverse research needs (Kalueff et al., 2014). Consequently, numerous experiments originally designed for rats have been successfully adapted to the zebrafish model.
Design and Functionality of the Zebrafish Y-Maze
The Zebrafish Y-Maze is structured as an aquarium in the shape of the letter ‘Y.’ Its three arms function as choice arms and a start arm, providing subjects with two distinct choices. During a baited aquatic Y-Maze task, subjects are expected to progress to the next arm after retrieving a reward from one arm in a trial. This simple behavioral task effectively assesses cognitive function by testing the subject’s ability to remember spatial locations, which relies on hippocampal-dependent reference memory. The Zebrafish Y-Maze serves as a valuable tool for evaluating learning and memory functions, as well as studying the impact of pharmacological interventions.
Design and Construction of the Zebrafish Y-Maze Aquarium
The Zebrafish Y-Maze aquarium, typically crafted in the shape of a symmetrical capital ‘Y,’ is predominantly made of glass or acrylic materials. Each of the three arms measures 25 cm in length and 8 cm in width, arranged at a 120-degree angle from one another. The tank’s height is approximately 15 cm.
Customization and Features
The construction materials can either be opaque or clear based on specific experimental requirements. Additional modifications, such as manually operated or automated guillotine doors, can be incorporated into each arm. These doors serve the purpose of either confining the fish within specific arms or creating novel spaces within the maze. Furthermore, the floor is designed in a contrasting color to the fish, facilitating easy observation of the fish’s movements and behaviors.
Care and Preparation of Zebrafish for Y-Maze Experiments
Ensuring the well-being of the subjects and maintaining optimal experimental conditions are critical for Y-Maze experiments involving zebrafish. Here are the key considerations for housing and preparing the fish:
Tank Conditions and Water Quality Maintenance
Y-Maze Tank Setup and Lighting
Behavioral Tracking and Video Recording
Acclimatization and Stress Reduction
Utilization of the Transparent Aquatic Y-Maze for Zebrafish Experimentation
In this experimental setup, a transparent aquatic Y-Maze is employed to conduct behavioral assessments. Visual cues in the form of geometric white paper cut-outs (circles, squares, and triangles) are affixed on the exterior walls of the maze arms. Each of the three arms is randomly assigned as the start arm, the novel arm, and the other arm.
Trial Procedure and Memory Evaluation
The task comprises of two trials. In the first trial, the novel arm is sealed off using a guillotine door. The fish is introduced to the start arm and allowed to explore the two open arms for 5 minutes. Subsequently, the guillotine door is gradually lifted over a minute, enabling the fish to explore all three arms for an additional 5-minute session.
Upon the establishment of preferential exploration of novelty, memory assessment is conducted at 1, 3, and 6 hours of training-test intervals (TTI). For the second trial, the fish is reintroduced to the same starting arm and permitted to explore all three arms for 5 minutes following various TTIs. It’s noteworthy that subjects are exposed to the novel arm only once, as different groups of fish are assessed in each interval. Furthermore, to randomize the cues, the maze is rotated for every experiment.
In the study, the zebrafish subjects were classified into two groups to assess the engagement of the glutamatergic and cholinergic systems in the acquisition and consolidation of recognition memory. The first group was exposed to MK-801 (5, 10, or 20 µM) for 15 minutes or scopolamine (50, 100, or 200 µM) for 1 hour before or immediately after the training session. Meanwhile, the control group remained in their original compartment of the home tank. Both treated and untreated subjects underwent testing as outlined in the Y-Maze task protocol.
The data collected from the Zebrafish Y-Maze includes
The sample graph shows the time spent in each arm as a response to novelty in the Y-maze task.
Another sample graph depicts the total distance traveled by the subject on treatment with MK-801.
The Zebrafish Y-Maze capitalizes on the innate exploratory behavior of the fish to evaluate spatial preference without involving conditioned learning. This aquatic maze provides a platform for specific memory testing and is particularly suitable for continuous alternation tasks, typically necessitating minimal training and testing time. The incorporation of guillotine doors aids in minimizing fish handling, thereby reducing stress. Furthermore, the natural turn angles within the maze contribute to mitigating motivational and emotional stress. Following the establishment of preferential exploration of novelty, memory assessment can be conducted over longer training test intervals, enhancing the scope of the evaluation.
Cognato, G., Bortolotto, J., Blazina, A., Christoff, R., Lara, D., Vianna, M., and Bonan, C., (2012). Y-Maze memory task in zebrafish (Danio rerio): the role of glutamatergic and cholinergic systems on the acquisition and consolidation periods. Neurobiol. Learn. Mem. 98(4): 321-8. doi: 10.1016/j.nlm.2012.09.008.
Conrad, C., Galea, L., Kuroda, Y., McEwen, B. (1996). Chronic Stress Impairs Rat Spatial Memory on the Y Maze, and This Effect Is Blocked by Tianeptine Pretreatment. Behav. Neurosci. 110(6): 1321-34. PMID: 8986335.
Kalueff AV, Stewart AM, Gerlai R (2014). Zebrafish as an emerging model for studying complex brain disorders. Trends Pharmacol Sci. 35(2):63-75. doi: 10.1016/j.tips.2013.12.002.
Van der Borght, K., Havekes, R., Bos, T., Eggen, B., Van der Zee, E. (2007). Exercise improves memory acquisition and retrieval in the Y-maze task: relationship with hippocampal neurogenesis. Behav. Neurosci. 121(2): 324-34. DOI: 10.1037/0735-7044.121.2.324
Yamazaki, K., Yamaguchi, M., Baranoski, L., Bard, J., Boyse, E., Thomas, L. (1979). Recognition among mice. Evidence from the use of a Y-maze differentially scented by congenic mice of different major histocompatibility types. J. Exp. Med 150(4): 755-60.
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