Non-human primate animal learning and reversion ability testing method

Abstract

The invention discloses a non-human primate animal learning and reversion ability testing method. According to the invention, a standardized test method and a test device are used for measuring the learning discrimination and reversion ability of non-human primates, changes of brain functions and behaviors of animals are evaluated more comprehensively, a model evaluation means can be provided for central nervous system diseases such as Alzheimer's disease and cerebral apoplexy, and a new method is provided for disease research and new drug research.

Description

Technical Field

[0001] The present invention belongs to the field of neurobiology, and particularly relates to a method for testing the learning and reversal ability of non-human primates. Background Art

[0002] In the field of neurobiology, it is often necessary to conduct behavioral tests on experimental animals. By having the experimental animals complete certain behavioral paradigms, their cognitive ability, learning ability, and brain function are evaluated, thereby revealing the laws of brain activity and providing evaluation methods for nervous system disease models (such as Alzheimer's disease model, Parkinson's disease model, etc.). As a high-level experimental animal, non-human primates have a highly complex nervous system and can complete a variety of complex behavioral tasks; evaluating their advanced brain function and cognitive function by completing these behavioral tasks is a very important research method in neurobiology.

[0003] Due to the relatively close genetic relationship between non-human primates and humans, non-human primate models play an indispensable and important role in studying the mechanisms of human diseases and developing new drugs. There is a need in the laboratory to measure the cognitive, memory, learning, and reversal abilities of non-human primates as an important means for disease model evaluation and drug efficacy evaluation.

[0004] Most of the existing behavioral evaluation methods for non-human primates are to evaluate their memory ability, cognitive ability, social behavior, and hand movement ability. There are few methods and corresponding instrument devices for evaluating their learning discrimination and reversal ability. Different from lower animals such as mice, non-human primates have a highly complex nervous system and have certain learning ability and reasoning ability. Therefore, establishing a method that can quantify the learning discrimination and reversal ability of non-human primates can more comprehensively evaluate the changes in animal brain function and behavior. Using standardized test methods and test devices to measure the learning discrimination and reversal ability of non-human primates can provide model evaluation means for central nervous system diseases such as Alzheimer's disease and stroke, and provide new methods for disease research and new drug research.

[0005] Defects of the prior art:

[0006] Most of the existing behavioral evaluation methods for non-human primates are to evaluate their memory ability, cognitive ability, social behavior, and hand movement ability. There are few methods and corresponding instrument devices for evaluating their learning discrimination and reversal ability.

[0007] The existing non-human primate behavioral paradigms include:

[0008]

[0009]

[0010] Defects of the existing reversal learning paradigm: Reference: ROBERT R. ZIMMERMANN. Reversal Learning in the Developing Malnourished Rhesus Monkey. BEHAVIORAL BIOLOGY, 8, 381 - 390 (1973), Abstract No. 1 - 184R.

[0011] 1. The paradigm design is too simple and does not fully consider the adaptability problem and visual confusion problem of monkeys.

[0012] 2. No adaptive training is set. Monkeys need to first adapt to actions such as the drop - board in the training cage, the flip - top on the well, and finding food in the well (that is, realizing that there is food reward in the well that can be taken). Placing food on the left or right well of the animal at the beginning and letting it feed will inevitably lead to an increase in the error rate.

[0013] 3. The number of times for training to reach the standard is set too simply. Monkeys reach the standard after 10 consecutive correct times. The number of times is too small, and actually the monkeys have not fully learned.

[0014] 4. Setting 10 different discrimination learning and reversal problems at one time is likely to confuse monkeys.

[0015] 5. The data processing only compares the number of errors. As the number of learning times increases, the error rate of monkeys itself will decrease. Simply comparing the number of errors cannot fully reflect the learning discrimination and reversal ability of monkeys.

[0016] 6. Using an older WGTA training cage with a smaller cage body is not conducive to animal welfare and the operation convenience of staff. Summary of the Invention

[0017] The purpose of the present invention is to provide a method for testing the learning and reversal ability of non - human primates. The present invention uses a standardized testing method and testing device to measure the learning discrimination and reversal ability of non - human primates, more comprehensively evaluate the changes in animal brain function and behavior, and can provide a model evaluation means for central nervous system diseases such as Alzheimer's disease and stroke, and provide new methods for disease research and new drug research.

[0018] A method for testing the learning and reversal ability of non - human primates according to the present invention includes the following steps:

[0019] A training cage is manufactured. The training cage includes a first cage body and a second cage body connected to each other. A support frame is provided between the first cage body and the second cage body. Slideways are provided on the edges of the support frames on both sides. An opaque board is provided between the first cage body and the second cage body. The two sides of the opaque board are inserted into the slideways, so that the opaque board separates the first cage body and the second cage body. The second cage body is sequentially provided with an observation cage, a training plate placement layer, a food placement layer, and a leg placement space from top to bottom. A one-way visible glass is provided on the front wall of the observation cage, and a camera for observing the situation inside the observation cage is provided below it. A tray for placing feces and urine is provided below the first cage body. A hole is provided on the upper edge of the support frame, and a hook is provided on the right edge of the support frame. A rope is fixed on the upper side of the opaque board. The rope passes through the hole on the upper edge of the support frame, and a weight is hung at the end. The weight can be stuck on the hook. A plurality of universal wheels with lock catches are provided below the training cage;

[0020] The first cage body is for placing animals, and the second cage body is for placing the training plate and the camera;

[0021] During training, people sit on the outside of the second cage body. A one-way visible glass is installed in front of the cage body. The staff observes the training plate and the animal situation through this glass, and performs operations such as placing food and dropping the plate;

[0022] The training plate placement layer of the second cage body is used to place the training plate and the food for training;

[0023] Both discrimination training and reversal training use two training plates. The present invention requires that the distance between the two training plates be consistent with the animal to avoid the animal being more inclined to choose the food closer to itself and reducing the factors affecting the training result.

[0024] The training plate is like a spoon, with a concave middle;

[0025] I. Preparation before the experiment:

[0026] a. The monkeys are raised by the fixed breeder. The monkeys have no excessive reaction after the breeder enters the monkey house;

[0027] b. During the breeding process, the breeder puts the food in his hand and guides the monkeys to reach out and take the food to establish a trust relationship;

[0028] c. During the breeding, find 2-3 kinds of food that each monkey likes the most for subsequent training rewards;

[0029] IV. Delayed response training

[0030] a. Before the experiment starts, the animals should be fasted. Select 2-3 kinds of food that the monkeys like as training rewards and cut them into small pieces;

[0031] b. Start the delayed response training. First, place the monkey in the first cage. After the monkey calms down, turn on the camera and perform the training operations according to the delayed response training form. After the training is over, turn off the camera and transfer the monkey back to the monkey house;

[0032] b.1 Delayed response process:

[0033] b.1.1 Transfer the monkey to the first cage, place the training food and the training form properly, pull down the delay board, open the lid of the food access hole. After the monkey calms down, start the delayed response training;

[0034] b.1.2 Place the food in the food access hole according to the position of the food well specified in the training form, put down the delay board, and after the delay times of 0 seconds, 1 second, 2 seconds... pull up the delay board, and let the monkey freely take the food. Record whether the food is taken correctly. If the wrong lid is opened, cancel the reward food in front of the monkey's eyes;

[0035] b.1.3 Delay time: The value of B starts to increase from 0. A, B, C, D, and E are 0, 1, 2, 3, and 4 times the value of B respectively. That is, when the value of B is 1s, for the first training "RA" in the form, the food is placed in the food well on the right side of the trainer, and the board - dropping time is 0 times the value of B, that is, 1s×0 = 0s; for the third training "RC", the food is placed in the food well on the right side of the trainer, and the board - dropping time is 2 times the value of B, that is, 1s×2 = 2s;

[0036] b.1.4 Repeat the operation and conduct 30 trainings every day according to the training form;

[0037] b.1.5 Train every day until the monkey is proficient in the training equipment, the operation of the lifting board, and the operation of opening the lid to take food;

[0038] V. Learning reversal adaptation training

[0039] a. Place the monkey in the first cage, turn on the camera, and start the experiment after the monkey calms down;

[0040] b. Place a food well in the middle of the operation table in front of the monkey;

[0041] c. Put down the opaque baffle, place the rewarded food in the food well, cover it with the lid of color A, pull up the baffle, wait for the monkey to take the food and then drop the baffle. Repeat the above steps 30 times. If the monkey can correctly obtain the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, the next - step training can be carried out the next day. Otherwise, repeat this training;

[0042] IV. Color discrimination training

[0043] a. Place the monkey in the first cage, turn on the camera, and start the experiment after the monkey calms down.

[0044] b. Assemble the two food wells together.

[0045] c. Lower the baffle, place the food in the food wells according to the table, cover the lid of color B on one side, cover the lid of color C on the opposite side, pull up the baffle, wait for the monkey to take the food (if the wrong food well is selected, the reward will be cancelled) or cancel the reward if the monkey does not take the food after 60 s, and then lower the baffle. Repeat this step 30 times.

[0046] d. If the monkey correctly obtains the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, then conduct reverse training on the same afternoon or the next morning, with an interval of no more than 12 hours. Otherwise, repeat this training.

[0047] VI. Reverse Training

[0048] a. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down.

[0049] b. Lower the baffle, place the food in the food wells according to the table, cover the lid of color C on one side, cover the lid of color B on the opposite side, pull up the baffle, wait for the monkey to take the food, cancel the reward if the wrong food well is selected or cancel the reward if the monkey does not take the food after 60 s, and then lower the baffle; repeat this step 30 times.

[0050] c. During this process, if the monkey fails to obtain the rewarded food for three consecutive sub - trainings, a rewarded food can be fed to the monkey with forceps, and then the training can continue.

[0051] d. If the monkey correctly obtains the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, then the reverse training is successful and the training stops. Otherwise, repeat this training.

[0052] Preferably, there is a feces and urine tray at the bottom of the first cage body, which can be pulled out for cleaning, and is locked with a buckle between the tray and the cage body to prevent the monkey from reaching under the cage to move the feces and urine tray playfully.

[0053] Preferably, there is a buckle 4 at the upper end of the door of the cage body for placing animals. Correspondingly, there is a hole at the upper end of the door of the animal import and export. After the monkey enters the cage, the door is lowered and the buckle is pushed out to effectively lock the door. There is a depression at each of the two edges of the middle and lower parts of the door. When the door is lifted, there is also a buckle at the corresponding position at the upper end of the cage body. Pushing out the buckle can effectively fix the door and prevent it from falling.

[0054] Preferably, for the opaque board between the first cage body and the second cage body, two pulleys, a traction rope and a weight are used to pull it up and down. And there is a downward hook on the side of the cage body to which the rope can be hooked, so as to fix the board at the desired height. The two pulleys can reduce the weight pressure of the opaque board on the technician, enabling the technician to easily pull up the board quickly, achieve precise control of the pulling - board time during training, and reduce errors.

[0055] Preferably, universal wheels with locks are used at the bottoms of both the first cage body and the second cage body.

[0056] Preferably, the training tray is made of stainless steel.

[0057] Preferably, there is also a lid on the training tray, which is an opaque acrylic board or a cup, or other materials such as plastic, stainless steel, etc. By setting different colors or different shapes, the objects that the monkeys need to distinguish can be set.

[0058] The training tray uses a mortise and tenon structure. One side protrudes as a semi-cylindrical shape, and the other side has a semi-cylindrical depression. It can be used alone or two or more can be assembled together. When assembling, no screws are required for fixation, and it is firmly assembled and also convenient for disassembly.

[0059] Further preferably, the lid on the training tray is made of stainless steel, circular, and there are two screws at one end that can be connected to the training tray.

[0060] Advantages:

[0061] The present invention uses standardized test methods and test devices to measure the learning discrimination and reversal abilities of non-human primates, more comprehensively evaluate the changes in animal brain functions and behaviors, can provide model evaluation means for central nervous system diseases such as Alzheimer's disease and stroke, and provide new methods for disease research and new drug research.

[0062] The present invention sets up a delayed response training process, which can enable monkeys to adapt to the training device and get used to the dropping plate of the device, so as to avoid affecting the animal's mood due to the reduction of the visible space after the dropping plate. The set delayed response training process can also enable monkeys to fully learn to reach out and flip the lid to take food from the food hole in the training cage.

[0063] The present invention sets up a link for screening the preferred foods of animals, screening out two or three kinds of foods that animals like to eat, and fixing these kinds of foods in the subsequent training, so as to avoid monkeys not taking food because they don't like to eat, increasing the error rate. Moreover, if there are two or three kinds of foods to choose from, it can also avoid the monkeys getting tired of eating only one kind of food during training and increasing the error rate. As one of the links of the standardized test method, this design can make the training results more scientific and reliable.

[0064] The present invention sets the size of the food to be consistent, 1 cm * 1 cm * 1 cm, to avoid the influence of different food sizes on the animal's desire to eat during training.

[0065] The present invention sets up adaptation training. Only one food well is placed, and the monkey is not allowed to see when the food is put in. By trying to flip the lid, the monkey can know that there is food reward in the well. The lid used in this adaptation training is different from the lid used in the subsequent learning discrimination and reversal training, so as to avoid confusion in training at different stages before and after. The adaptation training is set up before the formal training to make the learning process of the monkey gradual, so that the errors in the subsequent learning discrimination and reversal training come from the learning and cognitive errors of the monkey, rather than the cognitive errors of the device and method, making the training results more scientific and reliable.

[0066] The present invention sets the training standard as follows: 30 times of training each time, that is, if the correct rate is above 90%, the next step of training can be carried out the next day. This is to avoid too few times, and the monkey has not fully learned yet, and reaching the correct rate requirement is only accidental.

[0067] The present invention sets the reversal training to be carried out the day after the color discrimination training reaches the standard, and the interval does not exceed 12 hours, so as to avoid too long a time interval after completing the learning discrimination training, which fails to play the role of contrast during reversal. Different interval times may lead to different correct rates during reversal on the first day. Stipulating the interval time between the two kinds of training can ensure the reproducibility of this experiment.

[0068] During the training process of the present invention, the time interval from the falling plate to the rising should be controlled between 8 - 10 seconds, and there is a 10 - 15 - second pause between each time, so as to avoid the monkey being confused due to continuous training.

[0069] For the data processing of the present invention, four data are compared, namely the number of days and the number of training times required when the learning discrimination reaches the standard, and the number of days and the number of training times required when the reversal training reaches the standard. It can not only analyze the ability of learning discrimination, but also analyze the ability of learning reversal. Compared with only comparing the error rate, the data analysis is more in - depth. Because the ability of learning discrimination and the ability of learning reversal are not complementary. Some animals have good learning discrimination ability, but they are very difficult to react when reversing.

[0070] The training plate (food well) used in the present invention uses a way similar to the mortise - and - tenon structure. It can be used alone, or two or more can be assembled together. When assembling, no screws are needed for fixation. It is firmly assembled and also convenient for disassembly. It can be used for delayed response 2 - hole training, or multi - hole training, and can also be used for the learning discrimination and reversal training of the present invention paradigm.

[0071] The training plate (food well) used in the present invention is made of stainless steel material and has a relatively large weight. The monkey cannot easily pick it up, so as to avoid the monkey moving or overturning the food plate mischievously.

[0072] The training trays (food wells) are of the same size and depth, avoiding the situation where the food well is too deep and difficult to access, which may affect the monkey's desire to obtain food, or the food well is too shallow and the food cannot be completely blocked after covering the lid. Moreover, the food wells are of the same size, which, combined with the consistent size of the food (such as apple pieces), can improve the reproducibility of the experiment.

[0073] The lid on the food well can be an opaque acrylic board or a cup, or other materials such as plastic or stainless steel. By setting different colors or shapes, objects that the monkey needs to distinguish can be set up. It can be used for training in paradigms such as delayed response, learning discrimination, learning discrimination and reversal.

[0074] When necessary, the lid of the food well is made of stainless steel, circular, and has two screws at one end for connecting to the food well. When the monkey flips open the lid to get food, it can prevent the monkey from being naughty and playing with the lid.

[0075] Advantage 1 of the training cage: Compared with other training cages, the training cage used in the present invention consists of two cage bodies. The first cage body is for placing animals, and the second cage body is for placing the training tray and the camera. The second cage body is made of opaque acrylic board and can be tightly connected to the first cage body through a lock, which can effectively prevent the animal from being affected by the periphery. During training, the animal can only see the training tray and is more focused. The two cage bodies can also be disassembled and used separately for other experimental needs.

[0076] Advantage 2 of the training cage: During training, people sit outside the second cage body. A one-way visible mirror is installed in front of the cage body. The staff observes the training tray and the animal situation through this mirror and performs operations such as placing food and dropping the board. The animal cannot see the staff, which can prevent the animal from being affected during training and make the training more focused. It can also effectively prevent the staff from giving hints to the animal and interfering with the training results in order to improve the training accuracy and reduce working hours.

[0077] Both discrimination training and reversal training use two training trays. The present invention requires that the distance between the two training trays and the animal be the same, avoiding the animal being more inclined to choose the food tray closer to itself and reducing the factors affecting the training results.

[0078] Advantage 3 of the training cage: There is a platform for installing a camera under the mirror. From this angle, the training situation of the animal and the training tray can be photographed from the middle front. If the camera is installed in the corner, the distance in the video will be smaller in the distance and larger in the near distance. When others watch the video, they will think that the distance between the training tray and the animal is inconsistent. By photographing from the middle front, the video can accurately restore the training scenario.

[0079] Advantages of the training cage 4: The second cage body has three layers, and the height of each layer is different. The bottom layer is relatively spacious, which is convenient for the trainer to place both feet and will not get tired even after long-term training. The middle layer is the narrowest and is used to place training food (peanut grains, apple grains, etc.). Its height is the same as the height of the trainer's hand when sitting, and the hand will not get tired during training. The top layer is the training space. The width of the bottom layer is wider than the upper two layers, and there is a protruding part at the place where the food is placed, which is convenient for the trainer to get the food.

[0080] Advantages of the training cage 5: There is a feces and urine tray at the bottom of the first cage body, which can be pulled out for cleaning. There is a buckle between the tray and the cage body to lock it, preventing the monkey from reaching under the cage to move the feces and urine tray when being naughty. The height of the feces and urine tray is closely connected to the cage, and there are basically no gaps, preventing urine from splashing everywhere outside the cage when the monkey urinates.

[0081] Advantages of the training cage 6: There is a buckle at the upper end of the cage body for placing animals. Correspondingly, there is a hole at the upper end of the door of the animal import and export. After the monkey enters the cage and puts down the door, the buckle can be pushed out to effectively lock the door. There is a depression on each side edge of the middle and lower parts of the door. When the door is lifted, there is also a buckle at the corresponding position at the upper end of the cage body. Pushing out the buckle can effectively fix the door and prevent it from falling. The technician doesn't need to hold the door while driving the monkey from the transfer cage into the training cage, effectively liberating the technician's hands and facilitating the work.

[0082] Advantages of the training cage 7: For the opaque board between the animal cage body and the training cage body, two pulleys, a leash, and a weight are combined to pull it up and down. And there is a downward hook on the side of the cage body where the rope can be hung on the hook, so that the board can be fixed at the desired height. The two pulleys can reduce the weight pressure of the opaque board on the technician, enabling the technician to easily pull up the board quickly, achieving precise control of the pulling time of the board during training and reducing errors.

[0083] Advantages of the training cage 8: Both the bottom ends of the training cage body and the animal cage body use lockable universal wheels, which are convenient for movement and can also be easily fixed during use to prevent the monkey from shaking the cage and affecting training when being naughty. Description of the Drawings

[0084] Figure 1 is a schematic structural diagram of the training cage;

[0085] Figure 2 is a schematic structural diagram of the training tray (food well), where A is the training tray (food well) and B is the lid. Specific Implementation Method

[0087] The following embodiments are further descriptions of the present invention rather than limitations on the present invention.

[0088] As Figure 1As shown in the figure, the training cage of the present invention includes a first cage body 5 and a second cage body 15 connected to each other (for example, connected by a latch 8). A support frame 3 is provided between the first cage body and the second cage body. Slideways are provided on the edges of the support frames on both sides. An opaque plate 2 is provided between the first cage body and the second cage body. Both sides of the opaque plate are snapped into the slideways, so that the opaque plate separates the first cage body and the second cage body. The second cage body is sequentially provided with an observation cage 1, a training plate placement layer 12, a food placement layer 11, and a leg placement space 10 from top to bottom. The front wall of the observation cage is provided with a one-way visible glass 14, and a camera 13 for observing the situation inside the observation cage is provided below it. A tray 7 for placing feces and urine is provided below the first cage body. A hole is provided on the upper edge of the support frame, and a hook 16 is provided on the right edge of the support frame. A rope 17 is fixed on the upper side of the opaque plate. The rope passes through the hole on the upper edge of the support frame, and a weight 6 is hung at the end. The weight can be stuck on the hook 16. A plurality of universal wheels 9 with latches are provided below the training cage.

[0089] The training cage used in the present invention is composed of two cage bodies. The first cage body is for placing animals, and the second cage body is for placing the training plate and the camera. The second cage body is made of opaque acrylic board and can be tightly connected to the first cage body through a latch, which can effectively prevent animals from being affected by the outside. During training, animals can only see the training plate and are more focused. The two cage bodies can also be disassembled and used separately, which can be used for other experimental needs.

[0090] During training, people sit on the outside of the second cage body. A one-way visible glass is installed in front of the cage body. The staff observes the training plate and the animal situation through this glass and performs operations such as placing food and dropping the board. The animal cannot see the staff, which can prevent the animal from being affected during training and make the training more focused. It can also effectively prevent the staff from giving animals hints and artificially interfering with the training results in order to improve the training accuracy and reduce the working time.

[0091] Both discrimination training and reversal training use two training plates. The present invention requires that the distance between the two training plates is the same as that of the animal, so as to avoid the animal being more inclined to choose the food plate closer to itself and reduce the factors affecting the training results.

[0092] A platform for installing the camera is provided below the glass. From this angle, the training situation of the animal and the training plate can be photographed from the middle front. If the camera is installed in the corner, the distance in the video will become smaller in the distance and larger in the near distance. When others watch the video, they will think that the distance between the training plate and the animal is inconsistent. By shooting from the middle front, the video can accurately restore the training scene.

[0093] The second cage body has three layers, and the height of each layer is different. The bottom layer is relatively spacious, facilitating the trainer to place both feet, and the trainer will not feel tired even after long-term training. The middle layer is the narrowest and is used to place training food (peanut grains, apple grains, etc.). Its height is the same as the height of the trainer's hand when sitting, and the hand will not feel tired during training. The top layer is the training space. The width of the bottom layer is wider than the upper two layers, and there is a protruding part where the food is placed, facilitating the trainer to pick up the food.

[0094] At the bottom of the first cage body, there is a feces and urine tray that can be pulled out for cleaning. There is a buckle between the tray and the cage body to lock it, preventing the monkey from reaching out under the cage to move the feces and urine tray playfully. The height of the feces and urine tray is closely connected to the cage, with basically no gap, preventing urine from splashing everywhere outside the cage when the monkey urinates.

[0095] At the upper end near the door of the cage body for placing animals, there is a buckle 4. Correspondingly, there is a hole at the upper end of the door of the animal import and export. After the monkey enters the cage and closes the door, the buckle 4 can be pushed out to effectively lock the door. There is a depression on each of the two edges at the middle and lower parts of the door. When the door is lifted, there is also a buckle at the corresponding position at the upper end of the cage body. Pushing out the buckle can effectively fix the door and prevent it from falling. The technician doesn't need to hold the door while driving the monkey from the transfer cage into the training cage, effectively liberating the technician's hands and facilitating the work.

[0096] For the opaque board between the animal cage body (the first cage body) and the training cage body (the second cage body), two pulleys, a tow rope, and a weight are combined to lift and lower it. And there is a downward hook on the side of the cage body where the rope can be hung on the hook, enabling the board to be fixed at the desired height. The two pulleys can reduce the weight pressure of the opaque board on the technician, enabling the technician to easily lift the board quickly, achieving precise control of the pulling time of the board during training and reducing errors.

[0097] Both the bottom ends of the training cage body and the animal cage body use universal wheels with locks, which are convenient for moving and can also be easily fixed during use to prevent the monkey from shaking the cage playfully and affecting training.

[0098] As Figure 2 shown, the training tray (food well) used in the present invention is made of stainless steel, with a relatively large weight, and the monkey cannot easily pick it up, preventing the monkey from moving or overturning the food tray playfully.

[0099] The training tray (food well) is like a spoon, with a concave middle part. Its size and depth are consistent, avoiding the situation that the food well is too deep and difficult to pick up, which affects the monkey's desire to obtain food, or the food well is too shallow and cannot completely cover the food after covering the lid. Moreover, the size of the food wells is consistent, matching the size of the food (such as apple pieces) to improve the reproducibility of the experiment.

[0100] The lid on the food well can be an opaque acrylic board or a cup, or other materials such as plastic, stainless steel, etc. By setting different colors or different shapes, the objects that the monkey needs to distinguish can be set. It can be used for training in paradigms such as delayed response, learning to distinguish, learning to distinguish and reversal.

[0101] When necessary, the lid of the food well is made of stainless steel, circular, with two screws at one end for connection to the food well. When the monkey flips open the lid to get food, it can prevent the monkey from being naughty and taking the lid to play.

[0102] Example 1:

[0103] I. Preparation before the experiment:

[0104] 2. The feeder who needs to be fixed raises the monkeys. After the feeder enters the monkey house, the monkeys have no excessive reaction.

[0105] 3. During the feeding process, the feeder puts the food in the hand and guides the monkey to reach out and take the food to establish a trust relationship.

[0106] 4. During the feeding, find 2-3 favorite foods for each monkey for subsequent training rewards.

[0107] II. Delayed response training

[0108] 1. Before the experiment starts, the animals should be fasted. Select 2-3 of the monkey's favorite foods as training rewards and cut them into

[0109] small cubes of 1 cm * 1 cm * 1 cm.

[0110] 2. Start the delayed response training. First, place the monkey in the first cage body. After the monkey calms down, turn on the camera and perform the training operations according to the delayed response training form. After the end, turn off the camera and transfer the monkey back to the monkey house.

[0111] 2.1 Delayed response process:

[0112] 2.1.1. Transfer the monkey to the first cage body, place the training food and the training form, pull up the delay board, open the lid of the food-taking hole. After the monkey calms down, start the delayed response training.

[0113] 2.1.2. Place the food into the food-taking hole according to the position of the food well specified in the training form, lower the delay board, and after a delay time of 0 seconds, 1 second, 2 seconds... pull up the delay board, and let the monkey freely take the food. Record whether the food is taken correctly. If the wrong lid is flipped open, cancel the reward food in front of the monkey's eyes.

[0114] 2.1.3. Delay time: The B value increases from 0. A, B, C, D, and E are 0, 1, 2, 3, and 4 times the B value respectively. That is, in the first training in the table, the food is placed in the food well on the right side of the trainer, and the dropping board time is 0 s; the third training is that the food is placed in the food well on the right side of the trainer, and the dropping board time is 2 s (when the B value is 1 s).

[0115] 2.1.4. Repeat the operation and conduct 30 trainings every day according to the training table.

[0116] 2.1.5. Train every day until the monkeys are proficient in the training equipment, the operation of the lifting board, and the operation of opening the flap to obtain food.

[0117] 2.1.6. The training table is as follows:

[0118]

[0119]

[0120] III. Learning reverse adaptation training

[0121] 5. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down.

[0122] 6. Place a food well in the middle of the operation table in front of the monkey.

[0123] 7. Lower the opaque baffle, place the rewarded food in the food well, cover it with the lid of color A. Pull up the baffle, wait for the monkey to obtain the food and then lower the baffle. Repeat the above steps 30 times. If the monkey can correctly obtain the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, the next step of training can be carried out the next day, otherwise repeat this training.

[0124] IV. Color discrimination training

[0125] 2. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down.

[0126] 3. Assemble two food wells together.

[0127] 4. Lower the baffle, place the food in the food well according to the table, cover the lid of color B, and cover the lid of color C on the opposite well. Pull up the baffle, wait for the monkey to obtain the food (if the wrong food well is selected, the reward will be cancelled) or if the monkey does not obtain the food after 60 s, cancel the reward and lower the baffle. Repeat this step 30 times.

[0128] 5. If the monkey correctly obtains the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, conduct reverse training on the same afternoon or the next morning (within 12 hours), otherwise repeat this training.

[0129] V. Reverse Training

[0130] 1. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down.

[0131] 2. Lower the baffle, place the food in the food well according to the table, cover it with the C-color lid, cover the opposite well with the B-color lid, raise the baffle, and wait for the monkey to take the food (if the wrong food well is selected, the reward will be cancelled) or cancel the reward if the monkey does not take the food after 60 s, then lower the baffle. Repeat this step 30 times.

[0132] 3. During this process, if the monkey fails to obtain the rewarded food for three consecutive sub-training sessions, a rewarded food can be fed to the monkey with forceps and then the training can continue.

[0133] 4. If the monkey correctly obtains the rewarded food 27 times in 30 training sessions, that is, the correct rate is over 90%, the reverse training is successful and the training stops. Otherwise, repeat this training.

[0134] VI. Precautions

[0135] 1. The food should be selected as fresh and regular monkey food without irritation and addiction.

[0136] 2. The breeding and training environment should be kept comfortable and dry.

[0137] 3. The food intake should be appropriately reduced 12 hours before the behavioral training.

[0138] 4. During the training process, the time interval from lowering the board to raising it should be controlled between 8 - 10 s.

[0139] 5. Operations that intimidate or attack the experimental animals and affect their emotions are prohibited.

[0140] 6. During the training process, if the monkey does not take the food 60 s after being allowed to take the rewarded food, this sub-training session is regarded as a failure, the rewarded food is cancelled and the next sub-training session starts (if the monkey lifts the wrong one, it must not be allowed to get the food).

[0141] 7. When cancelling the reward, the food should be taken out and removed in front of the monkey so that the monkey knows which food well is the correct one.

[0142] 8. The color discrimination and reverse experiments should not be stopped, and the training should also be carried out on Saturdays and Sundays so as not to affect the state of the monkeys after two days.

[0143] 9. There should be a 10 - 15 s pause between each time, and continuous training is not allowed to avoid confusing the monkeys.

[0144] VII. Data Processing

[0145] After the training is completed, based on the data such as the number of training sessions and the number of training days required for the monkeys to distinguish colors and reverse training respectively collected, the learning ability of the monkeys can be preliminarily evaluated. Training is carried out again at the specified time points after the successful establishment of the model, and the postoperative indicators are collected (since this training is a learning behavior, no adaptive training is required again), and whether there are significant differences between the control group and the model group and before and after the establishment of the model is compared to analyze the impact of the model establishment on the behavior of the monkeys.

[0146] VIII. Recording Table

[0147]

[0148]

[0149] IX. Specific Implementation Case:

[0150] 1. Animals: 8 cynomolgus monkeys, age: 8 - 12 years old.

[0151] 2. Experimental Purpose:

[0152] To prepare a disease model monkey of Alzheimer's disease (AD) and apply the learning reversal training paradigm as one of the model evaluation indicators.

[0153] 3. Experimental Procedure: See II, III, IV, V.

[0154] 4. Experimental Results:

[0155]

[0156] Note: Original Learning - Color Discrimination Training; Reversal Learning - Reversal Training

[0157]

[0158]

[0159] As can be seen from the above table, the number of training sessions required for Monkey No. 1 to reach a certain standard in color discrimination and reversal training both increase after drug administration to establish the model, indicating that its learning ability and reversal ability are declining.

Claims

1. A method for testing the learning and reversal ability of non-human primates, characterized in that, Including the following steps: Manufacture a training cage, which includes a connected first cage body and a second cage body. There is a support frame between the first cage body and the second cage body. Slideways are provided on the edges of the two support frames. An opaque board is provided between the first cage body and the second cage body. Both sides of the opaque board are snapped into the slideways, so that the opaque board separates the first cage body and the second cage body. The second cage body is sequentially provided with an observation cage, a training plate placement layer, a food placement layer and a leg placement space from top to bottom. The front wall of the observation cage is provided with a one-way visible glass, and a camera for observing the situation inside the observation cage is arranged below it. A tray for placing feces and urine is arranged below the first cage body. A hole is provided on the upper edge of the support frame, and a hook is provided on the right edge of the support frame. A rope is fixed on the upper side of the opaque board. The rope passes through the hole on the upper edge of the support frame, and a weight is hung at the end. The weight can be stuck on the hook. A plurality of universal wheels with latches are arranged below the training cage; The first cage body is for placing animals, and the second cage body is for placing training plates and cameras; During training, people sit on the outside of the second cage body. A one-way visible glass is installed in front of the cage body. The staff observes the training plate and the animal situation through this glass, and performs operations such as placing food and dropping the board; The training plate placement layer of the second cage body is used to place the training plate and the food for training; Both discrimination training and reversal training use two training plates. The present invention requires that the distance between the two training plates is consistent with the animal to avoid the animal being more inclined to choose the food closer to itself and reducing the factors affecting the training results. The training plate is like a spoon, with a concave in the middle; I. Preparation before the experiment: a. The keeper who needs to be fixed raises the monkeys. After the keeper enters the monkey house, the monkeys have no stress response; b. During the feeding process, the keeper puts the food in his hand and guides the monkeys to reach out and take the food to establish a trust relationship; c. During the feeding, find 2-3 kinds of food that each monkey likes the most for subsequent training rewards; II. Delayed response training a. Before the experiment starts, the animals should be fasted. Select 2-3 kinds of food that the monkeys like as training rewards and cut them into small pieces; b. Start the delayed response training. First, place the monkeys into the first cage body. After the monkeys calm down, turn on the camera and perform training operations according to the delayed response training form. After the end, turn off the camera and transfer the monkeys back to the monkey house; b.1 Delayed response process: b.1.1 Transfer the monkeys to the first cage body, place the training food and the training form, pull up the delay board, open the lid of the food well, and start the delayed response training after the monkeys are quiet; b.1.2 Place the food into the food well according to the position of the food well specified in the training form, lower the delay board, and pull up the delay board after a delay time of 0 second, 1 second, 2 seconds... Let the monkeys take the food freely, record whether they take the food correctly. If the wrong lid is opened, cancel the reward food in front of the monkeys' eyes; b.1.3 Delay time: The value of B increases from 0. A, B, C, D, and E are 0, 1, 2, 3, and 4 times the value of B respectively. That is, when the value of B is 1 s, for the first training "RA" in the table, the food is placed in the food well on the right side of the trainer, and the dropping plate time is 0 times the value of B, that is, 1 s × 0 = 0 s; for the third training "RC", the food is placed in the food well on the right side of the trainer, and the dropping plate time is 2 times the value of B, that is, 1 s × 2 = 2 s; b.1.4 Repeat the operation and conduct 30 trainings per day according to the training table; b.1.5 Train every day until the monkey is proficient in the training equipment, the operation of the lifting plate, and the operation of opening the flap to obtain food; III. Learning Reverse Adaptation Training a. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down; b. Place a food well in the middle of the operation table in front of the monkey; c. Lower the opaque baffle, place the rewarded food in the food well, cover it with the lid of color A, pull up the baffle, wait for the monkey to obtain the food and then lower the baffle. Repeat the above steps 30 times. If the monkey can correctly obtain the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, the next step of training can be carried out the next day. Otherwise, repeat this training; IV. Color Discrimination Training a. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down. b. Assemble two food wells together. c. Lower the baffle, place the food in the food well according to the table, cover the well with the lid of color B, and cover the opposite well with the lid of color C. Pull up the baffle and wait for the monkey to obtain the food (if the wrong food well is selected, the reward will be cancelled) or if the monkey does not obtain the food after 60 s, the reward will be cancelled, and then lower the baffle. Repeat this step 30 times. d. If the monkey can correctly obtain the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, reverse training will be carried out on the same afternoon or the next morning, with an interval of no more than 12 hours. Otherwise, repeat this training; V. Reverse Training a. Place the monkey in the first cage body, turn on the camera, and start the experiment after the monkey calms down; b. Lower the baffle, place the food in the food well according to the table, cover the well with the lid of color C, and cover the opposite well with the lid of color B. Pull up the baffle and wait for the monkey to obtain the food. If the wrong food well is selected, the reward will be cancelled or if the monkey does not obtain the food after 60 s, the reward will be cancelled, and then lower the baffle; repeat this step 30 times; c. During this process, if the monkey cannot obtain the rewarded food for three consecutive sub-trainings, a rewarded food can be fed to the monkey with forceps and then the training can continue; d. If the monkey can correctly obtain the rewarded food 27 times in 30 trainings, that is, the correct rate is over 90%, the reverse training is successful and the training stops. Otherwise, repeat this training.

2. The method according to claim 1, wherein There is a feces and urine tray at the bottom of the first cage body, which can be pulled out for cleaning. There is a buckle lock between the tray and the cage body to prevent the monkey from reaching out under the cage to move the feces and urine tray mischievously.

3. The method according to claim 1, wherein There is a buckle 4 at the upper end of the cage body for placing animals. Correspondingly, there is a hole at the upper end of the door of the animal inlet and outlet. After the monkey enters the cage and closes the door, the buckle is pushed out to effectively lock the door. There is a depression at each of the two edges of the middle and lower parts of the door. When the door is lifted, there is also a buckle at the corresponding position at the upper end of the cage body. Pushing out the buckle can effectively fix the door and prevent it from falling.

4. The method according to claim 1, wherein For the opaque plate between the first cage body and the second cage body, two pulleys, a tow rope, and a weight are used in combination to lift and lower it. And there is a downward hook on the side of the cage body where the rope can be hooked, so that the plate can be fixed at the desired height. The two pulleys can reduce the weight pressure of the opaque plate on the technician, enabling the technician to easily lift the plate quickly and achieve precise control of the pulling time of the training plate, reducing errors.

5. The method according to claim 1, wherein The bottom ends of the first cage body and the second cage body both use lockable universal wheels.

6. The method according to claim 1, wherein The described training plate is made of stainless steel.

7. The method according to claim 1, wherein The training plate uses the mortise and tenon structure. One side protrudes in a semi-cylindrical shape, and the other side has a semi-cylindrical depression. It can be used alone or two or more can be assembled and used together.

8. The method according to claim 1, wherein There is also a lid on the training plate, which is an opaque acrylic plate or a cup, or other materials such as plastic or stainless steel. By setting different colors or different shapes, the objects that the monkey needs to distinguish can be set.

9. The method according to claim 1, wherein The lid on the training plate is made of stainless steel, circular, and there are two screws at one end for connecting to the training plate.

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