Method for training animal dependent visual clue positioning reward ability by using animal behavior training and testing system

Abstract

The invention provides a method for training the visual clue-dependent positioning reward capability of animals by using an animal behavior training and testing system, and the system realizes that one set of system covers multiple types of experiments through the opening and closing logic of an access door of an adjusting arm, the reward region reward putting rule and the free conversion of visual clues. A visual clue-reward association learning scene is constructed, and animals are guided to establish memory association between clues and reward positions. A three-stage visual clue navigation and memory ability testing method is established, the memory retention ability associated with clue rewards is evaluated in the association stage, the spatial memory ability is independently evaluated in the blank stage, and the regulation and control mechanism of visual clues on animal spatial memory is analyzed in the non-association stage.

Description

Technical Field

[0001] This invention relates to the technical field of animal behavior experimental methods, specifically to a method for training animals' ability to locate rewards based on visual cues using an animal behavior training and testing system. Background Technology

[0002] Research on learning and memory relies on precise assessments of animals' spatial cognition, working memory, and referential memory. Mazes, as a classic behavioral paradigm, simulate the "information encoding-consolidation-retrieval" memory process, making them a core tool for analyzing neural mechanisms and disease models. Currently, mainstream mazes include: Y / T maze, Morris water maze, Barnes maze, double H-shaped maze, and cross maze.

[0003] Traditional radial (eight-arm) mazes rely solely on "remote environmental cues (such as laboratory wall arm markings)" and lack dedicated visual cues within the arms, making it impossible to separate the memory contributions of "global spatial navigation" and "local visual cue recognition." Furthermore, cues are uncontrollable (e.g., marker wear, light changes), resulting in poor experimental repeatability. Door opening and closing, and reward placement largely depend on manual operation (e.g., manually pushing gates, placing food pellets), exhibiting a certain response delay. The timing of reward placement (e.g., how long after the animal enters the arm before giving the reward) cannot be precisely controlled, presenting significant limitations. In contrast, the classic cross maze (usually a self-alternating maze) is a classic memory assessment device designed based on animals' "natural instinct to explore novel environments." Its core function is to test working memory (short-term information storage and updating ability). It typically features a cross-shaped layout of "central area + 4 equal-length channel arms (at 90° to each other)," with the channel arms mostly open (without door obstructions). It largely lacks actively designed visual cues, relying solely on "arm length / color (simple spraying)." The clues are of a single type (only color blocks) and lack standardization (such as stripe density and grid size are not quantified), making it impossible to study the relationship between "visual features (texture / contrast) and navigation behavior".

[0004] As neuroscience research advances towards a deeper coupling of behavior and neural mechanisms, simple behavioral observations are no longer sufficient to analyze brain circuit functions. When conducting precise observations such as in vivo electrophysiological experiments and two-photon imaging, any human intervention on the animals during the task (such as manually opening and closing maze doors or manually delivering rewards) interferes with the stability of neural signals and the spontaneity of behavior, severely affecting the accuracy of recording the activity of spatial coding neurons such as place cells and grid cells. Therefore, the fully automated design of the entire process, including automatic opening and closing of maze doors and precise triggering of reward pumps, is essential. Furthermore, automating equipment operation through systems such as LabVIEW can completely eliminate human interference, ensuring that animals complete visual cue navigation tasks (such as spatial decision-making based on customized visual cues like black and white stripes or grids) while maintaining the continuity and reliability of neural signal recordings. The introduction of visual cues not only breaks through the limitations of traditional mazes that rely on global spatial cues, but also specifically focuses on the association mechanism of "visual cognition-navigation memory," providing behavioral paradigm support for the precise analysis of the function of the visual-spatial memory circuit. Combined with in vivo electrophysiology and two-photon imaging technology, it is possible to capture changes in the encoding patterns of place cells in real time when animals respond to visual cues, achieving multi-dimensional synchronous analysis of "behavioral phenotype-neural activity-circuit mechanism." This is precisely the core technological integration requirement for neuroscience to move from "behavioral description" to "mechanism elucidation," and it is also a necessary path for in-depth exploration of the neural basis of visual-spatial navigation. Summary of the Invention

[0005] To overcome the problems of existing technologies, this invention provides a training and testing system and method for animal cue-dependent goal-oriented spatial tasks and spatial memory generalization behaviors. It conducts visual cognition-navigation memory related training and testing through customized visual cue specificity, while also being compatible with precise observation technologies such as in vivo electrophysiology and two-photon imaging. This enables simultaneous acquisition of "behavioral performance - neural signals" in animals during free movement, effectively improving the standardization of experimental procedures and data accuracy, and meeting the needs of in-depth research into visual-spatial navigation mechanisms in neuroscience research. This invention provides the following technical solutions: A training and testing system for animal cue-dependent goal-oriented spatial tasks and spatial memory generalization behaviors includes a maze. The maze consists of a central area and four channel arms connected to the central area, forming an enclosed space for animals to move within. An entrance door and an entrance door opening / closing sensor are provided at the connection between the channel arms and the central area. A reward area is provided at the end of the channel arms. Visual cue differentiation patterns are provided on both sides of the channel arms. A detection sensor is installed in front of the reward area.

[0006] Furthermore, the reward area is a water tank, and the liquid reward storage device is connected to a dropper via a peristaltic pump, which then enters the water tank.

[0007] Furthermore, the lower part of the maze has a hollow structure, and the entrance door is a downward-opening mechanical door.

[0008] Furthermore, the channel arm is equipped with displays on both sides, which display the visual cue differentiation patterns. The visual cues are standard black and white stripes, black and white checks, pure black backgrounds, or diagonal brick shapes.

[0009] Furthermore, both the entrance door switch sensor and the detection sensor are infrared sensors.

[0010] It also includes an analysis and control module and a data storage module. The analysis and control module receives and analyzes signals from various sensors, synchronously controls the opening and closing of the entrance door and the switching on and off of the peristaltic pump, transmits visual cue differentiation patterns to the display screen, and records data in real time. The data storage module is used to store the data transmitted by the analysis and control module and equipment operation log information.

[0011] A method for training animals' ability to locate rewards based on visual cues using an animal behavior training and testing system includes the following steps: Step 1: The analysis and control module projects black and white striped visual cues onto the displays of Channel 1 and Channel 2; and randomly projects black and white checkered patterns and pure black background visual cues onto the displays of Channel 3 and Channel 4, respectively, and records the task type as No. 1 and No. 2. Step two: Place the experimental animals in the central area of ​​the maze; Step 3: The analysis and control module controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage in that area; if the result is inconsistent, proceed to the next step. Step 7: Steps 4 to 6 are repeated in a loop within a preset time period. When the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close, and the experiment ends. The data from step eight is saved in the data storage module in the form of an Excel spreadsheet.

[0012] A method for testing animal navigation and memory abilities using a training and testing system includes the following steps: Step 1: The analysis and control module projects black and white striped visual cues onto the displays of Channel 1 and Channel 2; and randomly projects black and white checkered patterns and pure black background visual cues onto the displays of Channel 3 and Channel 4, respectively, and records the task type as No. 1 and No. 2. Step two: Place the experimental animals in the central area of ​​the maze; Step 3: The analysis and control module controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage in that area; if the result is inconsistent, proceed to the next step. Step 7 involves looping through steps 4 to 6 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 1 to 7 are repeated three times. Step 8: Set the task types to No. 1-3, No. 1-4, No. 2-3, No. 2-4, and No. 3-4 respectively, adjust the visual cues on the passage wall, repeat steps 1 to 7, repeat three times for each task type, and save the data in the data storage module in the form of an Excel spreadsheet. Step 9: Set the task type to No. 1 or No. 2, associate the rewards for No. 1 and No. 2 channel arms, and turn off the displays of all channel arms. Step 10: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 11: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 12: Determine if the channel arm being entered is consistent with the channel arm corresponding to the set task type number; if the result is consistent, the analysis and control module will open the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drip liquid reward into the liquid reward storage of that area; if the result is inconsistent, proceed to the next step. Step 13 involves looping through Step 11 and Step 12 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 9 to 13 are repeated three times. Step Fourteen: Set the task type to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively. Repeat Steps Nine to Thirteen three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet. Step 15: The analysis and control module sets the task type to No. 1 and No. 2. The rewards are associated with the No. 1 and No. 2 channel arms. The display patterns of the four channel arms are fixed and are displayed as follows: No. 1 arm is black and white stripes, No. 2 arm is black and white checkered pattern, No. 3 arm is pure black pattern, and No. 4 arm is diagonal brick pattern. Step Sixteen: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 17: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 18: Determine if the channel arm being entered is consistent with the channel arm corresponding to the set task type number; if the result is consistent, the analysis and control module opens the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drips liquid reward into the liquid reward storage of that area; if the result is inconsistent, proceed to the next step. Step 19: Repeat steps 15 and 16 within the preset time. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 15 to 19 are repeated three times. Step 20 sets the task type to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively. Steps 15 to 19 are repeated three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet.

[0013] By adopting the above technical solution, the present invention has the following beneficial effects: 1. The training and testing system for goal-oriented spatial tasks and spatial memory generalization behavior of the present invention has high adaptability to experimental scenarios: By adjusting the "opening and closing logic of the arm's channel door, reward rules for reward zone, and free transformation of visual cues", the system can flexibly adapt to various behavioral experimental scenarios (such as goal-oriented spatial memory tasks based on visual cues, spatial working memory generalization tests, etc.), without the need for additional equipment replacement, achieving "one system covering multiple types of experiments", and significantly reducing experimental costs and equipment space occupation.

[0014] 2. The accuracy and objectivity of the behavioral testing method of the present invention: Each arm passage door is equipped with a switch sensor and the reward area is equipped with an infrared detection sensor, which can collect core behavioral data such as the animal's "entry / exit sequence and stay time in the reward area" in real time and without contact, avoiding subjective errors of manual observation; the analysis and control module automatically and synchronously records sensor signals and door operation commands, forming a closed-loop data chain of "behavior-control", ensuring the traceability and accuracy of experimental data.

[0015] 3. Non-intrusiveness of the animal behavior testing method of the present invention: The maze door adopts a mechanical structure of "up and down moving opening and closing", combined with the hollow design of the lower part of the maze, the door movement will not protrude from the inner arm of the passage, completely avoiding the physical obstruction of the door to the animal's passage and ensuring the naturalness of its behavior. 4. High efficiency in operation and data management: The analysis and control module of this invention integrates the functions of "signal analysis, gate control, and data recording", and runs automatically throughout the process without the need for real-time manual intervention; the data storage module uniformly retains experimental data, supports subsequent batch analysis, and the modules communicate stably through serial ports to avoid the risk of data transmission interruption or loss. 5. Flexibility in functional expansion: The visual cues at both ends of the maze arms of this invention can be freely replaced, which can better fit more refined behavioral experiments such as visual cue navigation training and cue association memory testing, providing more diverse technical support for exploring animal cognitive mechanisms. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the maze structure in an embodiment of the present invention; Figure 2 This is a schematic diagram of visual cues on both sides of the maze arm in an embodiment of the present invention; Figure 3 This is a flowchart of the method for training animal goal-oriented spatial memory according to the present invention; Figure 4 This is a flowchart of the method for testing the visual cue navigation and memory abilities of animals according to the present invention; Figure 5 This is a diagram showing the experimental results of the present invention.

[0017] The components are: 1. Central Area; 2. No. 1 Channel Arm; 6. No. 1 Channel Entrance Door; 10. No. 1 Channel Entrance Door Sensor; 18. No. 1 Arm Reward Area; 22. No. 1 Arm Detection Sensor; 26. No. 1 Arm Visual Cue; 3. No. 2 Channel Arm; 7. No. 2 Arm Channel Entrance Door; 11. No. 2 Arm Channel Entrance Door Sensor; 19. No. 2 Arm Reward Area; 23. No. 2 Arm Detection Sensor; 27. No. 2 Arm Visual Cue; 4. No. 3 Channel Arm; 8. No. 3 Arm Channel Entrance Door; 12. No. 3 Arm Channel Entrance Door Sensor; 20. No. 3 Arm Reward Area; 24. No. 3 Arm Detection Sensor; 28. No. 3 Arm Visual Cue; 5. No. 4 Channel Arm; 9. No. 4 Arm Channel Entrance Door; 13. No. 4 Arm Channel Entrance Door Sensor; 21. No. 4 Arm Reward Area; 25. No. 4 Arm Detection Sensor; 29. ​​No. 4 Arm Visual Cue; 30. Example of In-Arm Visual Cue; 31. Baffle. Detailed Implementation

[0018] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the structural diagrams and specific embodiments described herein are only for explaining the invention and are not intended to limit the invention. Example 1

[0019] Figure 1 is a schematic diagram of the structure of a maze according to the present invention. As shown in Figure 1, a training and testing system for animal cue-dependent goal-oriented spatial tasks and spatial memory generalization behavior includes a maze. The maze is a closed space formed by a central area 1 and four channel arms connected to the central area, accommodating animal movement within it. Four baffles 31 are provided at the edges of the central area that are not connected to the arms to prevent animals (such as mice) from escaping from the maze. The four channel arms are channel arm 1 (2), channel arm 2 (3), channel arm 3 (4), and channel arm 4 (5). Entrance gates are provided at the connection points between the channel arms and the central area, namely entrance gate 6 for channel arm 1, entrance gate 7 for channel arm 2, entrance gate 8 for channel arm 3, and entrance gate 9 for channel arm 4. Each entrance gate can be opened or closed to control the connection between the central area of ​​the maze and the corresponding reward area within the arm, controlling the movement of experimental animals between the central area and the arms / between arms. Reward zones are located at the ends of the passage arms: Reward Zone 18 (Arm 1), Reward Zone 19 (Arm 2), Reward Zone 20 (Arm 3), and Reward Zone 21 (Arm 4). Differentiated visual cues are located on both sides of the passage arms: Visual Cue 26 (Arm 1), Visual Cue 27 (Arm 2), Visual Cue 28 (Arm 3), and Visual Cue 29 (Arm 4). Infrared signal detection sensors are installed in front of the reward zones: Detection Sensor 22 (Arm 1), Detection Sensor 23 (Arm 2), Detection Sensor 24 (Arm 3), and Detection Sensor 25 (Arm 4). Similarly, detection sensors are installed in front of each passage entrance door: Entrance Door Sensor 10 (Arm 1), Entrance Door Sensor 11 (Arm 2), Entrance Door Sensor 12 (Arm 3), and Entrance Door Sensor 13 (Arm 4). These sensors record infrared trigger signals, mark the arm number, entry time, reward time, and exit time.

[0020] The reward area is a water tank, and the liquid reward storage device is connected to a dropper via a peristaltic pump, which then enters the water tank. The lower part of the maze has a hollow structure, and the entrance door is a downward-opening mechanical door. When the door rises, it blocks the passage; when it falls, it is completely retracted into the hollow area at the bottom of the maze. This design avoids both protruding from the inner wall of the passage and creating physical obstacles for the experimental animals, as well as preventing the door's movement from startling the animals. This maximizes the smoothness and naturalness of the animals' movement within the passage.

[0021] The walkway arm has displays on both sides showing visually differentiated patterns, such as... Figure 2 As shown, in-arm visual cue example 30, the visual cue can be a standard black and white stripe, black and white checkered pattern, pure black background or diagonal brick shape. Figure 2Taking Arm 1, section 2 as an example, the visual cue configuration scheme on both sides of each arm channel is shown in detail. In target-oriented spatial tasks that rely on visual cues, this system is equipped with four differentiated visual cues as standard, specifically including black and white stripes, black and white checks, a pure black background, and diagonal brick-shaped patterns. Furthermore, these visual cues are primarily displayed on the screens at both ends of the arm. The screens can automatically change the displayed visual cue content according to experimental needs, completing the cue switching operation and fully adapting to different experimental scenarios related to visual cognition and navigation memory.

[0022] The entrance door switch sensor and detection sensor are both infrared sensors. The system includes an analysis and control module and a data storage module. The analysis and control module receives and analyzes signals from each sensor, synchronously controls the opening and closing of the entrance door and the switching on and off of the peristaltic pump, transmits visual cue differentiation patterns to the display screen, and records data in real time. The data storage module stores the data transmitted by the analysis and control module and equipment operation log information. The maze's control circuitry and power supply are integrated below the maze, and the modules interact and transmit commands via serial cables.

[0023] The animal behavior training and testing system of the present invention can achieve training and testing of various animal behavior models by adjusting the opening and closing logic of different arm passage doors and the rules for giving sucrose water in the reward area, such as experimental scenarios such as animal goal-oriented spatial task testing based on visual cues and animal spatial memory generalization training. Example 2

[0024] This invention provides a method for training animals' ability to locate rewards based on visual cues using the animal behavior training and testing system of this invention. For example... Figure 3 As shown, three visual cues were used during training: black and white stripes, black and white checks, and solid black. The black and white striped cue was consistently used as the associated visual cue for the reward arm, while the solid black and black and white checkered cues were randomly assigned to the non-reward arms. This constructed a "visual cue-reward" associative learning scenario, guiding the animals to establish a memory association between the cue and the reward location. The specific experimental steps are as follows: Step 1: The analysis and control module projects black and white striped visual cues onto the displays of Channel 1 and Channel 2; and randomly projects black and white checkered patterns and pure black background visual cues onto the displays of Channel 3 and Channel 4, respectively, and records the task type as No. 1 and No. 2. Step two: Place the experimental animals in the central area of ​​the maze; Step 3: The analysis and control module controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage device in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage device in that area; if the result is inconsistent, proceed to the next step. Step 7: Steps 4 to 6 are repeated in a loop within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close, thus ending the experiment. The data from step eight is saved in the data storage module in the form of an Excel spreadsheet.

[0025] This training method can set up six task types. Besides task types one and two (i.e., channels one and two are associated with liquid rewards, while channels three and four are not), task types can also be one-three, one-four, two-three, two-four, and three-four. The reward arm settings are associated with the corresponding task type. The operation steps and execution logic for the other five tasks are the same as those in the above embodiments, only the reward arm numbers for different task types are different. Throughout the training process, the black and white striped visual cue is always associated with the reward arm of the current task, while pure black or black and white checkered visual cues are randomly assigned to non-reward arms. The setting of visual cues is synchronously matched according to the switching of reward arms. Example 3

[0026] Figure 4 shows a method for testing visual cue navigation and memory abilities using an animal behavior training and testing system. The experiment consists of three core stages: The cue-reward association phase: A fixed association is maintained between the black and white striped cue and the reward arm. Plain black cues and black and white checkered cues are randomly assigned to non-reward arms. This aims to verify the animal's ability to retain the established cue-reward association. The method and steps are consistent with the animal training method for locating rewards based on visual cues in Example 2.

[0027] Blank test phase: All visual cues are completely removed, and the position of the reward arm is adjusted only according to the task type to eliminate visual cue interference and achieve independent assessment of the animal's spatial memory ability.

[0028] The cue-reward non-associated stage: Based on the three sets of visual cues, a new diagonal brick-shaped pattern cue is added. The four channel arms (channel arms one to four) are fixed to each set of visual cues (arm one: black and white stripes; arm two: black and white checkered pattern; arm three: pure black; arm four: diagonal brick shape). The position of the reward arm changes dynamically with the task type. This is used to assess the animal's navigation decision-making ability under the condition that the cue and reward are not fixedly associated, and then to analyze the regulatory mechanism of visual cues on the animal's spatial memory.

[0029] The specific experimental steps are as follows: Steps one through eight are the cue-reward association test; steps nine through fourteen are the blank test phase; and steps fifteen through twentieth are the cue-reward non-association phase. Step 1: The analysis and control module projects black and white striped visual cues onto the displays of Channel 1 and Channel 2; and randomly projects black and white checkered patterns and pure black background visual cues onto the displays of Channel 3 and Channel 4, respectively, and records the task type as No. 1 and No. 2. Step two: Place the experimental animals in the central area of ​​the maze; Step 3: The analysis and control module controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage in that area; if the result is inconsistent, proceed to the next step. Step 7 involves looping through steps 4 to 6 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 1 to 7 are repeated three times. Step 8: Set the task types to No. 1-3, No. 1-4, No. 2-3, No. 2-4, and No. 3-4 respectively. Adjust the visual cues of the passage wall and the association between the passage wall and the reward. Repeat steps 1 to 7 three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet. Step 9: Set the task type to No. 1 or No. 2, associate the rewards for No. 1 and No. 2 channel arms, and turn off the displays of all channel arms. Step 10: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 11: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 12: Determine if the channel arm being entered is consistent with the channel arm corresponding to the set task type number; if the result is consistent, the analysis and control module will open the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drip liquid reward into the liquid reward storage of that area; if the result is inconsistent, proceed to the next step. Step 13 involves looping through Step 11 and Step 12 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 9 to 13 are repeated three times. Step Fourteen: Set the task types to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively, adjust the association between the channel wall and the reward, repeat steps nine to thirteen, repeat three times for each task type, and save the data in the data storage module in the form of an Excel spreadsheet. Step 15: The analysis and control module sets the task type to No. 1 and No. 2. The rewards are associated with the No. 1 and No. 2 channel arms. The displays of the four channel arms show: No. 1 is black and white stripes, No. 2 is black and white checkered pattern, No. 3 is pure black pattern, and No. 4 is diagonal brick pattern. Step Sixteen: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 17: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 18: Determine if the channel arm being entered is consistent with the channel arm corresponding to the set task type number; if the result is consistent, the analysis and control module opens the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drips liquid reward into the liquid reward storage of that area; if the result is inconsistent, proceed to the next step. Step 19: Repeat steps 15 and 16 within the preset time. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 15 to 19 are repeated three times. Step 20 sets the task types to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively, adjusts the association between the channel wall and the reward, and repeats steps 15 to 19 three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet.

[0030] In step eight, black and white stripe visual cues are projected onto the display screens of the channel arms associated with the reward; black and white checkered patterns and pure black background visual cues are randomly projected onto the display screens of the unassociated channel arms, respectively. That is, when the task type is one or three, black and white stripe visual cues are projected onto the display screens of channel arms one and three; black and white checkered patterns and pure black background visual cues are randomly projected onto the display screens of channel arms two and four, respectively, for example.

[0031] In step fourteen, the displays of all four channel arms are turned off, and the channel wall selected for the task type is associated with the reward. That is, when the task type is one or three, channel arm one and channel arm three are associated with the reward.

[0032] In step 20, the display patterns on the four channel arms are fixed: arm 1 has black and white stripes, arm 2 has black and white checks, arm 3 has a pure black pattern, and arm 4 has a diagonal brick pattern. The channel wall selected for the task type is associated with the reward; that is, when the task type is one or three, channel arms 1 and 3 are associated with the reward. Example 4

[0033] like Figure 5 As shown, 10 mice were trained and tested using the method of this invention. The experimental parameters were: 8 days of training period, 6 task types per day (12, 13, 14, 23, 24, 34).

[0034] All experimental mice strictly followed the pre-set procedures of this invention and completed all tasks during the training and testing periods. From day 6 of the training period, the mice maintained a stable retention rate of over 70% for the "cue-reward association" (calculated as: number of correct entry into the reward arm / total number of tasks). During the testing period, mice could autonomously make navigation decisions based on visual cues or spatial memory, and significant differences were observed in the behavioral data across different testing phases. The accuracy rate increased with the number of training days, reflecting the gradual establishment of the "visual cue-reward arm" association memory during training. The learning curve showed a typical upward trend, indicating that mice could recognize and remember the visual signals associated with rewards through repeated attempts. The accuracy rate remained above 70% for at least two days, ruling out the possibility of accidental correct guesses and demonstrating the stability of this association memory—the mice not only learned to navigate using visual cues but could also stably apply this memory to subsequent task selections.

[0035] The above-mentioned training period experimental results fully demonstrate that the testing method described in this invention has a clear process design and well-defined operation steps, which can effectively guide experimental animals to complete the entire testing process. Moreover, the testing process is stable and controllable, which fully proves the feasibility of the method.

[0036] The embodiments described above are merely illustrative of implementation methods of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A method for training animals' ability to locate rewards based on visual cues using an animal behavior training and testing system, characterized in that, Includes the following steps: Step 1: The analysis and control module of the animal behavior training and testing system projects black and white striped visual cues onto the displays of the No. 1 and No. 2 channels; and randomly projects black and white checkered and pure black background visual cues onto the displays of the No. 3 and No. 4 channels, respectively, and records the task type as No. 1 and No.

2. Step 2: Place the test animal into the central area of ​​the maze of the animal behavior training and testing system. The maze is a closed space formed by the central area and four channel arms connected to the central area. The connection between the channel arm and the central area is equipped with an entrance door and an entrance door opening and closing sensor. At the end of the channel arm, there is a reward area. A detection sensor is installed in front of the reward area. The reward area is a water tank. The liquid reward storage device is connected to a drip tube through a peristaltic pump and is connected to the water tank through the drip tube. Step 3: The analysis and control module of the animal behavior training and testing system controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage device in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage device in that area; if the result is inconsistent, proceed to the next step. Step 7: Steps 4 to 6 are repeated in a loop within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the channel arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close, thus ending the experiment. The data from step eight is stored in the data storage module of the animal behavior training and testing system in the form of an Excel spreadsheet.

2. A method for testing visual cue navigation and memory ability using the animal behavior training and testing system of claim 1, wherein, Includes the following steps: Step 1: The analysis and control module of the animal behavior training and testing system projects black and white striped visual cues onto the displays of the No. 1 and No. 2 channels; and randomly projects black and white checkered and pure black background visual cues onto the displays of the No. 3 and No. 4 channels, respectively, and records the task type as No. 1 and No.

2. Step 2: Place the test animal into the central area of ​​the maze of the animal behavior training and testing system. The maze is a closed space formed by the central area and four channel arms connected to the central area. The connection between the channel arm and the central area is equipped with an entrance door and an entrance door opening and closing sensor. At the end of the channel arm, there is a reward area. A detection sensor is installed in front of the reward area. The reward area is a water tank. The liquid reward storage device is connected to a drip tube through a peristaltic pump and is connected to the water tank through the drip tube. Step 3: The analysis and control module controls all four entrance doors of the arms to open, allowing the experimental animals to explore the maze freely. Step 4: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 5: The analysis and control module determines whether the channel arm the animal enters is consistent with the channel arm corresponding to the current task type number; If the result of step six is ​​consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage device in the channel arm where the animal is located, and drip liquid reward into the liquid reward storage device in that area; if the result is inconsistent, proceed to the next step. Step 7 involves looping through steps 4 to 6 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 1 to 7 are repeated three times. Step 8: Set the task types to No. 1-3, No. 1-4, No. 2-3, No. 2-4, and No. 3-4 respectively. Adjust the visual cues of the passage wall and the association between the passage wall and the reward. Repeat steps 1 to 7 three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet. Step 9: Set the task type to No. 1 or No. 2, associate the rewards for No. 1 and No. 2 channel arms, and turn off the displays of all channel arms. Step 10: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 11: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 12: Determine whether the incoming channel arm matches the channel arm corresponding to the set task type number; If the judgment result is consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drip liquid reward into the liquid reward storage of that area; if the judgment result is inconsistent, proceed to the next step. Step 13 involves looping through Step 11 and Step 12 within a preset time period. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 9 to 13 are repeated three times. Step Fourteen: Set the task types to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively, adjust the association between the channel wall and the reward, repeat steps nine to thirteen, repeat three times for each task type, and save the data in the data storage module in the form of an Excel spreadsheet. Step 15: The analysis and control module sets the task type to No. 1 and No.

2. The rewards are associated with the No. 1 and No. 2 channel arms. The displays of the four channel arms show the following respectively: No. 1 arm is black and white stripes, No. 2 arm is black and white checkered pattern, No. 3 arm is pure black pattern, and No. 4 arm is diagonal brick pattern. Step Sixteen: The analysis and control module controls all four arm access doors to open, allowing the experimental animals to explore the maze freely. Step 17: Experimental animals randomly enter the reward area of ​​any channel arm. The entrance door switch sensor and the reward area detection sensor are triggered. The analysis and control module records the number of the animal entering the channel arm, the entry time, the reward time, and the time of leaving the channel arm. Step 18: Determine whether the incoming channel arm matches the channel arm corresponding to the set task type number; If the judgment result is consistent, the analysis and control module will activate the peristaltic pump corresponding to the liquid reward storage of the channel arm where the animal is located and drip liquid reward into the liquid reward storage of that area; if the judgment result is inconsistent, proceed to the next step. Step 19: Repeat steps 15 and 16 within the preset time. Once the experimental time reaches the set time and the entrance door switch sensor detects that the experimental animal has left the arm and returned to the central area, the analysis and control module controls all entrance doors of the four arms to close. Steps 15 to 19 are repeated three times. Step 20 sets the task types to No. 13, No. 14, No. 23, No. 24, and No. 34 respectively, adjusts the association between the channel wall and the reward, and repeats steps 15 to 19 three times for each task type. The data is saved in the data storage module in the form of an Excel spreadsheet.

3. The method according to claim 1 or 2, characterized in that, The lower part of the maze has a hollow structure, and the entrance door is a downward-opening mechanical door.

4. The method according to claim 1 or 2, characterized in that, The channel arm is equipped with displays on both sides, which display the visual cue differentiation patterns. The visual cues are standard black and white stripes, black and white checks, pure black background, or diagonal brick shapes.

5. The method according to claim 1 or 2, characterized in that, Both the entrance door switch sensor and the detection sensor are infrared sensors.

6. The method of claim 1 or 2, wherein, The animal behavior training and testing system includes a maze, an analysis and control module, and a data storage module. The analysis and control module receives and analyzes signals from various sensors in the maze, synchronously controls the opening and closing of the entrance door and the switching on and off of the peristaltic pump, transmits visual cue differentiation patterns to the display screen, and records data in real time. The data storage module is used to store the data transmitted by the analysis and control module and equipment operation log information.

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