Non-human primate visual function and behavior detection method

Abstract

The invention belongs to the technical field of animal visual function detection, and discloses a non-human primate visual function and behavior detection method. The method comprises the following steps of: 1, carrying out experimental analysis on behaviors that animals find and grab food in a plurality of small grids in an environment of interaction with experimenters; and 2, carrying out climbing motion behavior analysis on primate animals in a single behavior device cage without being disturbed by people. According to the method, the phenomena of insufficient visual functions and abnormal behaviors of the primates can be accurately captured once or continuously for multiple times, and the behavior ability and the change condition of the visual functions of the primates are evaluated through digital processing. The method provided by the invention can effectively solve the problems of insufficient evaluation of visual functions and behavior defects of primates in conventional ophthalmology detection, provides more datamation parameters, can deeply analyze the visual functions and individual differences of each primate, and accurately reflects the visual functions and corresponding behavior changes thereof.

Description

Technical Field

[0001] The present invention belongs to the technical field of animal visual function detection, and more specifically, relates to a method for detecting visual function and behavior of non-human primates. Background Art

[0002] Animal behavior studies the behavioral characteristics of animals themselves and their behavioral manifestations in their interactions with the environment and other organisms. It focuses on distinguishing between physiological and pathological behavioral characteristics and changes in animals, and provides important data for the exploration of animal characteristics and behaviors and the study of animal disease models. Especially in the study of experimental animal disease models, studying the behavior of animals can effectively reflect the experimental results and the evidence that matches the research conclusions. Experimental non-human primates (such as crab-eating macaques, rhesus monkeys, and marmosets) have significant advantages as experimental animals in terms of human genetics, physiology, reproduction, and ethics. These advantages make non-human primates indispensable experimental animals in medical and life science research. In the future, with the continuous development of science and technology, the application prospects of primates in experimental research will be broader. Hereditary retinopathy is a common genetic variation disease with a complex pathogenesis that seriously threatens human visual function and behavioral ability. It has become a research hotspot in the field of ophthalmology in recent years. Since clinical trials of such diseases have certain limitations, it is of far-reaching significance to establish an animal model that is adapted to them and then explore the mechanism of retinal genetic variation-related diseases. Since primates cannot express visual problems in words like humans, conventional ophthalmic function testing equipment must be used to obtain visual-related structural and functional data from primates under anesthesia. Repeated anesthesia and testing will not only cause certain repeated damage to the animals, but also fail to evaluate the specific situation of visual function in actual applications. Conventional ophthalmic examinations are not comprehensive in detecting animal visual function and ignore certain compensatory behavioral functions of visual function. At present, there is no effective behavioral evaluation method or digital data on visual function for non-human primate disease models produced by spontaneous, induced, and genetically altered hereditary retinopathy, which limits the study of visual function. Summary of the invention

[0003] In order to make up for the deficiencies in the prior art, the present invention proposes a method for detecting the visual function and behavior of non-human primates based on feeding and characteristic movements.

[0004] The second object of the present invention is to provide application of the above method.

[0005] The purpose of the present invention is achieved through the following technical solutions:

[0006] A method for testing the visual function of a non-human primate comprises a first experiment, wherein the first experiment comprises the following steps:

[0007] S1. Place the test device in front of the non-human primates in the test group, so that the animals can put their hands deep into the test device;

[0008] S2. Record the behavior of the animals in the following tests, which are: testing the lighting patterns of different environments, and randomly placing the animals' favorite objects in the test device, recording the time from the placement of the objects to the animals grabbing the objects, the number of times the animals reach out per unit time, the degree of the animals' eye gaze, and the effect of the object color preference on the animals' grabbing;

[0009] S3. The above behavioral performances are used as test indicators of the visual function of non-human primates to examine the visual function of non-human primates.

[0010] Preferably, in the above method, the lighting modes described in step S2 are 1000 lux and above, 500 lux, 250 lux, 125 lux and below respectively.

[0011] Preferably, in the above method, the testing device described in step S1 is a food box with a plurality of grids, the food box is made of acrylic, and the size of each grid in the food box is between 1.0 and 3.0 cm.

[0012] More preferably, the method further comprises a second experiment, wherein the second experiment is to place the non-human primate in a test cage with variable height, and record the behavior of the animal in different lighting modes and different cage heights, wherein the behavior comprises:

[0013] (1) The time it takes for the animal to climb up or down the cage once;

[0014] (2) the animal's climbing speed and number of climbs in the cage;

[0015] (3) The number of jumps per minute of the animal in the cage.

[0016] Preferably, in the second experiment, the lighting mode is 1000 lux and above and 250 lux and below.

[0017] Specifically, the operation of the second experiment is to record the behavior of animals in the test cage under the illumination modes of 1000 lux and above and 250 lux and below respectively; including the following steps:

[0018] (1) Lighting mode is 1000 lux and above

[0019] S1. Place the non-human primate in a test cage with variable height, adjust the cage height to the first height, allow the animal to move freely without human disturbance, and then record the animal's behavior for 15 minutes;

[0020] S2. Adjust the cage height to the second height. After the animals move freely without human disturbance, record the animals' behavior for 15 minutes.

[0021] S3. Adjust the cage height to the third height. After the animals move freely without human disturbance, record the animal's behavior for 15 minutes.

[0022] (1) The illumination mode is 250 lux or less, and S1 to S3 are repeated.

[0023] As a specific implementation, the total height of the test cage is, for example, 1500 mm, and is provided with two partitions, the two horizontal partitions being inserted into the climbing cage in the form of inserts. Initially, the animal is placed in the first partition space from the bottom up, the first height of the test cage is, for example, 500 mm, when the first partition from the bottom up is taken out, the second height of the test cage is, for example, 1000 mm, when the second partition from the bottom up is taken out, the third height tested is, for example, 1500 mm.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The method of the present invention can accurately capture the visual function deficiency and abnormal behavior of primates once or continuously, and evaluate the changes in the behavioral ability and visual function of primates through digital processing. The method invented by us can effectively make up for the problem of insufficient evaluation of visual function and behavioral defects of primates in conventional ophthalmological examinations, and provide more data parameters. It can also deeply analyze the visual function and personalized differences of each primate, and accurately reflect the visual function and its corresponding behavioral changes. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is an acrylic food box with 32 small squares used for feeding training;

[0027] Figure 2 To obtain food used for training;

[0028] Figure 3 It is a schematic diagram of the structure of a variable height climbing cage; DETAILED DESCRIPTION

[0029] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention will be further described below in conjunction with specific drawings and embodiments. In the embodiments, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used are all commercially available unless otherwise specified.

[0030] Example 1 Behavioral experiment based on random food grabbing by animals under interaction with experimenters

[0031] 1. Experimental apparatus and food

[0032] A customized 32-grid (the number can be adjusted) small square food box suitable for non-human primates of different sizes ( Figure 1 ), making it easier for animals to grab food with their hands.

[0033] The food box is made of transparent acrylic. The experimental food is colorful and animal-friendly food, such as raisins, banana chips, hawthorn chips, small biscuits, etc. ( Figure 2 ), the size is relatively uniform, and the grid size of the food box is between 1.0 and 3.0 cm, or adjusted according to the progress of the disease.

[0034] 2. Operation method

[0035] The above experimental device was used to conduct a non-human primate food grabbing experiment under different lighting conditions. The experiment was divided into four groups of lighting modes, and measured using a light meter, namely 1000 lux and above, 500 lux, 250 lux, and 125 lux and below, which roughly correspond to natural white light from 12:00 to 13:00 noon, natural light from 17:00 to 18:00 in the afternoon, dim natural light from 18:00 to 19:00 in the afternoon, and darkness after 19:00. In each lighting mode, food was randomly placed in one of the 32-grid food boxes, and moved to a fixed animal cage where the animal could reach out, waiting for the animal to grab the food.

[0036] 3. Record results and analysis

[0037] Record the time from placing the food to grabbing it, the number of times the hand reaches out (when the vision is abnormal, it is often difficult to successfully grab the food in one go, and it will grab it multiple times, and its wrong grabs are often around the correct grid), the degree of eye gaze and other performances, the influence of food color, etc. The experimental cycle is to pause for more than 10 minutes every 5 times. Due to the impact of food sugar content on animal health, it is recommended that the number of times of feeding should not exceed 25 times a day. Each group of each batch of experiments must complete more than 50 times. Comprehensively analyze the experimental data obtained.

[0038] like Figure 1In the statistical table of the experimental results of a certain experiment in Table 1, the one-year-old experimental group of crab-eating macaques M3 with abnormal vision and the control group of crab-eating macaques WT01 and WT02 were statistically analyzed for taking food from 32 small square food boxes, and key state information and characteristic reactions can be extracted. The control group of crab-eating macaques succeeded in taking food once under the first three lighting modes, and all completed it quickly and accurately, while the experimental group of crab-eating macaques with abnormal vision had an accuracy rate of 75% within 5 times under natural white light simulation, an accuracy rate of 45% within 5 times under dim natural light simulation from 17:00 to 18:00, and could not effectively take food under dim natural light simulation from 18:00 to 19:00. The experimental results are consistent with the characteristics of early-onset retinal degeneration, with worse vision in the dark and a continuous decline in vision with age.

[0039] Table 1

[0040] Cage grouping / status M3 manifestations of abnormal vision in the experimental group Control group WT01, WT02 White light / light Feeling + looking at the food box + 75% accuracy within 5 times Completed quickly and accurately Dim 17:00~18:00 Natural light Fumbling + Reduced eye gaze + 45% accuracy within 5 attempts Completed quickly and accurately Dim 18:00~19:00 Natural light Fumbling + not looking + accuracy close to 0 or meaningless Completed quickly and accurately Darkness after 19:00 Do not approach or eat Do not approach or eat

[0041] Example 2: Climbing behavior experiment of non-human primates in individual behavior apparatus cages without interruption

[0042] 1. Experimental setup and environment

[0043] A variable height climbing cage was customized, the total height of which can be set to 1500mm (the height can be adjusted according to the height characteristics of the animal) Figure 3 ), and is equipped with two partitions and three independent environments of different heights. Taking the height of 1500mm as an example, the adjustable height is 500mm, 1000mm, and 1500mm. The three sides of the cage are iron frames for non-human primates to climb, and one side is made of transparent acrylic material, which is convenient for filming and observation. The cage is placed in a ventilated laboratory without disturbing noise. Toys are placed in the cage. It is also equipped with a surveillance camera to record the animal's movement behavior. A speaker is configured to play music that simulates the natural environment or music that the animal is familiar with. The companion cage is placed on one side of the variable height climbing cage to reduce the anxiety of the experimental animals. The indoor illumination is set to two sets of lighting environments: 1000lux and above and 250lux and below, or adjusted according to the experiment.

[0044] In this embodiment, the climbing cage used has two horizontal partitions inserted into the climbing cage in the form of inserts. Initially, the animal is placed in the first partition space from the bottom to the top. The first height of the test cage is 500 mm. When the first partition is taken out from the bottom to the top, the second height of the test cage becomes 1000 mm. When the second partition is taken out from the bottom to the top, the third height becomes 1500 mm.

[0045] 2. Operation method

[0046] The above device was used to test the climbing behavior of primates under different lighting conditions. The experiment was divided into two groups of lighting environments and measured using a light meter, which were 1000 lux and above and 250 lux and below, respectively, which roughly corresponded to natural white light from 12:00 to 13:00 noon and natural light from 18:00 to 19:00 in the afternoon. The time or illumination can also be adjusted according to the experimental requirements. In each group of lighting environments, the animals were placed in a variable height cage. First, the cage height was adjusted to 500mm. In the mode of no human interference, the animals were allowed to move freely for 5 minutes before recording for 15 minutes. An adjacent partition was removed and the height was adjusted to 1000mm. In the mode of no human interference, the animals were allowed to move freely for 5 minutes before recording for 15 minutes. Another adjacent partition was removed and the height was adjusted to 1500mm. The recording operation was the same as above. The experiment was repeated for more than three groups at each height under each lighting environment. (For young animals, they will spin around quickly and chirp continuously when they first enter the cage. It is recommended to place the animals in the experimental cage and interact with them before the formal experiment to adapt, and then conduct the experiment after their emotions are stable.) Finally, analyze the recorded animal movement behavior video.

[0047] 3. Results Analysis

[0048] The behavior of animals in different lighting environments and cage heights was recorded, specifically quantified as the time it took for the animals to climb up or down the cage, the climbing speed and number of times, and the number of jumps per minute in the cage (normal primates often climb up and down quickly and jump multiple times in the cage, but when the vision is abnormal, the climbing speed and number of jumps are significantly reduced due to the inability to see the cage bars clearly, and the limbs have tentative groping movements when climbing). The standard for recording up and down climbing is that the animal's upper limbs hold the cage bars tightly, and the lower limbs are raised to start timing, and the lower limbs fall steadily to the platform. The standard for jumping is that the animal accumulates power and jumps from the original place to another place, and holds the cage bars tightly, which is recorded as one time.

[0049] Tables 2 and 3 show the statistical analysis of a climbing behavior experiment of a 1-year-old cynomolgus monkey in a variable height climbing cage. The 1-year-old control group cynomolgus monkey WT01 and the 1-year-old experimental group cynomolgus monkey M3 with abnormal vision were subjected to a climbing behavior experiment in a variable height climbing cage, and the video was recorded and analyzed. The results showed that the behavior characteristics of the control group cynomolgus monkey WT01 in the 500mm variable height climbing cage were fast spinning, exploring everywhere, and high activity. The time for climbing once in the 1000mm variable height climbing cage was 1 to 2 seconds, and the time for climbing once in the 1500mm variable height climbing cage was 2 to 3 seconds, and the number of jumps in the cage was 2 to 5 times / min, while the behavior characteristics of the experimental group cynomolgus monkey M3 with abnormal vision in the 500mm variable height climbing cage were slow spinning and sometimes squatting in the corner. The time for crawling in a 1000mm variable height climbing cage is 3 to 7 seconds, and the time for crawling in a 1500mm cage height is 5 to 13 seconds, with no jumping movements during the period, which is significantly different from the control group. The experimental results are consistent with the characteristics of early-onset retinal degeneration.

[0050] Table 2 Statistical analysis of the M3 high cage climbing exercise experiment of cynomolgus monkeys with abnormal vision in the experimental group

[0051]

[0052]

[0053] Table 3 Statistical analysis of the high cage climbing exercise experiment of the control group WT01

[0054]

[0055] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A method for testing the visual function of non-human primates, characterized in that: The method comprises a first experiment, wherein the first experiment comprises the following steps: S1. Place the test device in front of the non-human primates in the test group so that the animals can put their hands deep into the test device; S2. Record the behavior of the animals in the following tests, which are: testing the lighting patterns of different environments, and randomly placing the animals' favorite objects in the test device, recording the time from the placement of the objects to the animals grabbing the objects, the number of times the animals reach out per unit time, the degree of the animals' eye gaze, and the effect of the object color preference on the animals' grabbing; S3. The above behavioral performances are used as test indicators of the visual function of non-human primates to examine the visual function of non-human primates.

2. The method for testing the visual function of non-human primates according to claim 1, characterized in that: The illumination modes described in step S2 are 1000 lux and above, 500 lux, 250 lux, and 125 lux and below.

3. The method for testing the visual function of non-human primates according to claim 1, characterized in that: The testing device described in step S1 is a food box with a plurality of grids, the material of the food box is acrylic, and the size of each grid in the food box is between 1.0 and 3.0 cm.

4. A method for testing the visual function of non-human primates, characterized in that: The second experiment is to place non-human primates in a test cage with variable heights and record the behavior of the animals in different lighting modes and different cage heights, wherein the behavior includes: (1) The time it takes for the animal to climb up or down the cage once; (2) the animal's climbing speed and number of climbs in the cage; (3) The number of jumps per minute of the animal in the cage.

5. The method for testing the visual function of non-human primates according to claim 4, characterized in that: The lighting mode is 1000 lux and above and 250 lux and below.

6. The method for testing the visual function of non-human primates according to claim 5, characterized in that: The operation of the second experiment is to record the behavior of animals in the test cage under the lighting mode of 1000 lux and above and 250 lux and below respectively; Specifically, the steps include: (1) Lighting mode is 1000 lux and above S1. Place the non-human primate in a test cage with variable height, adjust the cage height to the first height, allow the animal to move freely without human disturbance, and then record the animal's behavior for 15 minutes; S2. Adjust the cage height to the second height. After the animals move freely without human disturbance, record the animals' behavior for 15 minutes. S3. Adjust the cage height to the third height. After the animals move freely without human disturbance, record the animal's behavior for 15 minutes. (1) The illumination mode is 250 lux or less, and S1 to S3 are repeated.

7. The method for testing the visual function of non-human primates according to claim 6, characterized in that: The test cage is set at three heights with a total height of 1500 mm, and is provided with two partitions, and the two horizontal partitions are inserted into the climbing cage in the form of inserts. Initially, the animal is placed in the first partition space from the bottom to the top, which is the first height of the test cage, 500 mm. When the first partition from the bottom to the top is taken out, it is the second height of the test cage, 1000 mm. When the second partition from the bottom to the top is taken out, it is the third height of the tested cage, 1500 mm.

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