$3,995.00
The Automated Avoidance Zebrafish Y-maze is an operant conditioning assay designed to study zebrafish avoidance behaviors in response to aversive stimuli. This setup includes a Y-maze with an LCD screen positioned underneath that displays visual cues throughout the maze.
This system eliminates the need for manual control of multiple stimuli, thereby reducing human error and saving time.
MazeEngineers provide the Automated Zebrafish Y-maze Avoidance Maze, which can be customized in terms of coloring and other features upon request.
Specifications |
Acrylic Y-maze with walls and translucent base |
Length: 15cm |
Width: 7cm |
Height: 10cm |
The Automated Avoidance Zebrafish Y-maze is an advanced operant conditioning assay designed to study zebrafish avoidance behaviors when exposed to aversive stimuli. It enhances the traditional Zebrafish Y-maze Avoidance setup, which relies solely on visual cues displayed via an LCD screen beneath the maze. The automated version incorporates additional stimuli, including auditory cues from speakers and aversive stimuli administered through electric shocks via steel mesh plates on the opposite walls of each maze arm. This system also includes video tracking software that manages the protocol, controlling the duration and timing of each stimulus, thereby reducing human error and saving time.
In this learning task, zebrafish must learn to associate a colored cue with an electric shock. All regions of the floor are colored red by the LCD screen except for one area designated as the “goal region,” which remains gray. The zebrafish must swim to the goal region within a set time limit (usually 15 seconds) to avoid an electric shock administered throughout the maze, except for the goal arm, until the trial ends. An auditory cue signals the start of each trial and can also serve as a conditioned stimulus alongside or instead of visual cues.
The Automated Avoidance Zebrafish Y-maze offers significant flexibility and can be easily adapted for various experimental protocols, eliminating the need for multiple behavioral apparatuses. For instance, it can be used for visual and auditory discrimination paradigms by placing different stimuli in each maze arm. This maze is valuable for studying zebrafish behaviors, which can be extrapolated to understand human behavioral phenotypes in behavioral neuroscience. Additionally, it aids in investigating the neural basis of behaviors, neuropsychiatric disorders, and the effects of drugs on the brain (Marcon, Benvenutti, Gallas-Lopes, Herrmann, & Piato, 2021).
The Automated Avoidance Zebrafish Y-maze features an acrylic structure with three arms arranged in a Y-shape, complemented by video tracking software. Each arm of the maze measures 15 cm in length, 7 cm in width, and 10 cm in height. A camera positioned 60 cm above the maze captures the behavior of the subjects. The video tracking software is responsible for setting up the experimental protocol and managing the experiment.
The maze includes various conditioning stimuli. An attached speaker delivers auditory stimuli with frequency tones ranging from 100 to 20,000 Hz and a volume scale from 1 to 100. Visual stimuli are presented on an LCD screen placed beneath the maze, controlled by specialized software. Mesh grids cover all maze walls, and steel mesh plates on both sides of the maze arms can deliver electric shocks. These shocks have a DC setting of 5V (0.12 mA), 50-ms duration, 10 Hz, and an AC setting of 2.5V. The voltage, current, and duration can be adjusted as needed.
The video tracking software regulates all stimuli, including auditory cues, visual signals, and electric shocks, all of which are customizable. The setup includes a control box connected to a PC that oversees all operations.
Separate the subjects by placing them individually in 1-liter tanks a day before the experiment.
On the day of the experiment, position a subject in the Automated Avoidance Zebrafish Y-Maze. Secure the maze with a transparent lid to prevent any escape. Ensure the entire floor of the tank remains colored gray (53 lx). Use the video tracking software to monitor the subject’s movements. Allow a habituation period of 10 minutes before beginning the trials.
This habituation period is essential to minimize the stress the subject might experience from being moved from its home tank to the experimental setup and to alleviate any fear associated with the new environment.
Conduct twenty initial trials without shocks, followed by one hundred training trials with shocks. Place a subject in the maze and begin each trial by presenting an auditory cue (700 Hz, 0.5 s, 63 dB). Designate one arm end as the goal region, ensuring it is an area the subject is not currently in or has not recently visited. Use the LCD screen to change the color of all regions except the goal region to red (27 lx). Maintain this color until the subject reaches the goal region or until 30 seconds have passed during the shock trials. Upon reaching the goal region, provide an auditory cue (1400 Hz, 0.5 s, 63 dB). If the subject does not reach the goal region within 15 seconds during the shock trials, administer a continuous electric shock to all regions of the maze except the goal arm until the trial ends.
During the 30-second inter-trial intervals, allow the apparatus floor to remain colored gray. This interval is crucial for allowing the subject to recover from the aversive experience.
Evaluation of associative learning in zebrafish using an Automated Avoidance Y-Maze
Aoki, Tsuboi, and Okamoto (2015) designed the Automated Avoidance Y-maze to investigate associative learning in zebrafish (Danio rerio) and enhance experimental efficiency. They used wild-type zebrafish aged 6-12 months. The maze was placed in a soundproof box atop a 24.1-inch LCD screen. The subjects’ behaviors were recorded with a video camera connected to a computer. The video software controlled auditory tones via a speaker and displayed visual cues on the screen. Electric shocks were administered through steel mesh plates in the maze arms. Additionally, small fans integrated into the screen maintained the water temperature between 29-30°C and dissipated heat from the screen.
The experiment began with twenty initial trials without shocks, followed by one hundred trials with shocks. During the training, the goal region was highlighted in red on the LCD screen, and the subject had to reach the goal within a set time limit (30 seconds for ‘no shock’ trials and 15 seconds for ‘shock’ trials). If the subject failed to reach the goal region during ‘shock’ trials, electric shocks were delivered throughout the maze except for the goal arm. Subjects exhibited active locomotion throughout all trials. Results showed that in the ‘no shock’ trials, subjects consistently reached the goal region within 15 seconds, though there was no initial preference for the goal arm. In the ‘shock’ trials, subjects achieved a 100% success rate in reaching the goal region by trials 41-43. However, between trials 44-61, there was a decline in success despite repeated shocks, with subjects only rushing to the goal region once the shock was administered. During trials 62-120, the success rate improved, with subjects swimming directly to the goal region. Overall, the findings indicated that the subjects demonstrated associative learning of electric shocks with red color cues and learned to exhibit avoidance behaviors in response to these cues.
The following parameters can be observed using the Automated Avoidance Zebrafish Y-Maze:
The Automated Avoidance Zebrafish Y-maze streamlines the experimental process by automating tasks typically done manually. It removes the need for constant monitoring of fish movement, activation of visual cues, auditory signals, and electric shocks, as well as deactivating all stimuli. The video tracking software manages the entire experiment and monitors the subject’s movement throughout. This automation reduces human error and negates the necessity for the experimenter to be present in the room, thereby minimizing potential influences on the learning process.
In contrast, manual execution of the experiment requires an observer to be in the room, which can affect the outcomes. The maze is equipped with configurable stimuli, including visual, auditory, and aversive elements, which can be adjusted for various experimental protocols. Additionally, using multiple apparatuses enables simultaneous training of multiple fish, further enhancing efficiency.
The size of the fish used in the maze is a crucial consideration, and the protocol may need to be adjusted accordingly. Smaller fish, for instance, might struggle to reach the goal region quickly, while larger fish might do so with minimal effort. Consequently, the size, age, and strain of the fish can all impact the results.
Additionally, the LCD screen beneath the tank can increase water temperature, so it’s essential to monitor this closely and use cooling fans to prevent overheating, which can stress the subjects. Visual cues should be kept at an appropriate brightness to avoid affecting the fish’s visual system. Repeated electric shocks can have significant mental and physical effects on the subjects, so it’s important to limit the number of trials conducted each day to a suitable amount.
Aoki, R., Tsuboi, T., & Okamoto, H. (2015). Y-maze avoidance: an automated and rapid associative learning paradigm in zebrafish. Neuroscience research, 91, 69–72. https://doi.org/10.1016/j.neures.2014.10.012
Marcon, M., Benvenutti, R., Gallas-Lopes, M., Herrmann, A. P., & Piato, A. (2021). What do zebrafish prefer? Directional and color preferences in maze tasks. bioRxiv. https://doi.org/10.1101/2021.12.22.473814
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