A method for detecting visual function and behavior in non-human primates

Abstract

The present application 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: 1. behavior experiment analysis of animals searching for and grabbing food in multiple small squares in an interactive environment with experimenters; and 2. primate climbing motion behavior analysis in a single behavior device cage without disturbance. The method of the present application can accurately capture the visual function deficiency and abnormal behavior of primates for a single time or multiple times in succession, and evaluate the behavior ability and visual function change of primates through digital processing. The method can effectively make up for the problems of conventional ophthalmic detection in evaluating the visual function deficiency and behavior defects of primates, provide more data parameters, deeply analyze the visual function and individual differences of each primate, and accurately reflect the visual function and corresponding behavior change.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of animal visual function detection, and more particularly to a non-human primate visual function and behavior detection method. BACKGROUND

[0002] Animal behavior research is the study of animal behavior characteristics and their interaction with the environment and other organisms, focusing on distinguishing between physiological and pathological behavior characteristics and changes, providing important data for animal behavior research and animal disease model research. In particular, in experimental animal disease model research, the behavior of the research animal can effectively reflect the experimental results and the evidence matching the research conclusions. Experimental non-human primates (such as cynomolgus monkeys, rhesus monkeys, and marmosets) have significant advantages in human genetics, physiology, reproduction, and ethics as experimental animals, making non-human primates an indispensable experimental animal in medical and life science research. With the continuous development of science and technology, the application of primates in experimental research will become more promising. Genetic retinal degeneration is a common genetic variation disease with complex pathogenesis, which seriously threatens human visual function and behavior, and has become a research hotspot in the field of ophthalmology in recent years. Due to the limitations of clinical trials for such diseases, it is of great significance to establish animal models adapted to genetic variation-related retinal diseases and to explore the mechanisms of such diseases. Since primates cannot express visual problems in language like humans, it is necessary to use conventional ophthalmic function detection equipment to obtain visual structure and function data under anesthesia. This repeated anesthesia and detection not only causes certain repeated damage to the animals, but also cannot evaluate the specific situation of visual function in actual application, and the conventional ophthalmic detection of animal visual function is not comprehensive, ignoring certain compensatory behavior functions of visual function. Currently, there is no effective behavior evaluation method and digital data acquisition of visual function for non-human primate disease models made by the three means of spontaneous, induced, and genetic changes related to genetic retinal degeneration, limiting the study of visual function. SUMMARY

[0003] In order to make up for the deficiencies in the prior art, the present application proposes a non-human primate visual function and behavior detection method based on feeding and characteristic movement.

[0004] A second object of the present application is to provide the use of the above method.

[0005] The object of the present application is achieved by the following technical solutions:

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

[0007] S1, place the test device in front of the non-human primates to be tested so that the animals can reach into the test device with their hands;

[0008] S2, record the behavior of the animals in the following tests, which are tests of different light modes and randomly placing items that the animals like in the test device, and recording the time from placing the items to the animals grabbing the items, the number of times the animals reach out per unit time, the degree of eye gaze of the animals, and the effect of color preference of the items on the animals' grabbing;

[0009] S3, the above behavior is used as a test index of the visual function of non-human primates to investigate the visual function of non-human primates.

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

[0011] Preferably, in the above method, the test device in step S1 is a feeding box with multiple grids, the material of the feeding box is acrylic, and the size of each grid in the feeding box is between 1.0 and 3.0 cm.

[0012] More preferably, the above method further comprises a second experiment, which is placing non-human primates in a test cage with variable height, recording the behavior of the animals in different light modes and different cage heights, and the behavior includes:

[0013] (1) the time taken by the animals to climb up or down the cage once;

[0014] (2) the climbing speed and frequency of the animals in the cage;

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

[0016] Preferably, in the second experiment, the light 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 the animals in the test cage under the light mode of 1000 lux and above, and 250 lux and below, respectively; including the following steps:

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

[0019] S1, place the non-human primates in a test cage with variable height, adjust the cage height to the first height, and record the behavior of the animals after they freely move in the mode without human disturbance, and the recording time is 15 minutes;

[0020] S2, adjust the cage height to the second height, after the animal is free to move in the mode without human disturbance, record the behavior of the animal, and the recording time is 15 min;

[0021] S3, adjust the cage height to the third height, after the animal is free to move in the mode without human disturbance, record the behavior of the animal, and the recording time is 15 min;

[0022] (1) the light mode is 250 lux and below, and S1-S3 is repeated.

[0023] As a specific embodiment, the total height of the test cage is, for example, 1500 mm, and two partitions are provided, and the two horizontal partitions are inserted into the climbing cage in the form of an insert piece. Initially, the animal is placed in the first partition space from bottom to top, and the first height of the test cage is, for example, 500 mm. When the first partition from bottom to top is removed, the second height of the test cage is, for example, 1000 mm. When the second partition from bottom to top is removed, the third height of the test is, for example, 1500 mm.

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

[0025] The method of the present application can accurately capture the visual dysfunction and behavior abnormality of primates for a single time or multiple times, and evaluate the behavior ability and visual function change of primates after digital processing. The method can effectively make up for the problem of evaluating the visual dysfunction and behavior defects of primates in conventional ophthalmic detection, and provide more data parameters, and can also deeply analyze the visual function and individual differences of each primate, and accurately reflect the visual function and the corresponding behavior change. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 is a 32-square acrylic food box used for feeding training;

[0027] Figure 2 is food used for feeding training;

[0028] Figure 3 is a schematic view of a variable-height climbing cage structure; DETAILED DESCRIPTION

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

[0030] Example 1: Behavior experiment of animal randomly grabbing food under interaction with experimental personnel

[0031] 1. Experimental device and food

[0032] A 32-cell (the number can be adjusted) small square food box (the size of the animal can be adjusted) is customized for different sizes of non-human primates, which is convenient for animals to grab food with their hands. Figure 1

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

[0034] 2. Operation method

[0035] Using the above experimental device, non-human primates were used to grab food under different light conditions. The experiment was divided into 4 groups of light modes, and a light meter was used to measure the light intensity, which was 1000 lux and above, 500 lux, 250 lux, 125 lux and below, which can be roughly corresponding to natural white light at 12:00-13:00, natural light at 17:00-18:00, dim natural light at 18:00-19:00, and darkness after 19:00. In each light mode, one of the 32 cells in the food box was randomly placed with food, and the animal cage was moved to the front of the fixed hand, and the animal grabbed the food.

[0036] 3. Record results and analysis

[0037] Record the time from placement to grabbing food, the number of times to stretch out the hand (it is often difficult to grab food once when vision is abnormal, and it will be grabbed multiple times, and the wrong grab is often around the correct grid), the degree of eye fixation and other performance, the influence of food color, etc. The experimental period is 10 minutes or more for every 5 times, and due to the influence of food sugar on animal health, it is recommended that the number of food intake per day should not exceed 25 times. Each batch of experiments needs to complete 50 times or more in each group. The obtained experimental data is comprehensively analyzed.

[0038] For example Figure 1 ​​and Table 1, the 1-year-old experimental group vision abnormality of cynomolgus monkeys M3 and control group cynomolgus monkeys WT01, WT02 were taken to the 32 small square food box to extract the key state information and characteristic reaction. The control group of cynomolgus monkeys once successfully fed in the first three groups of light mode, and all completed quickly and accurately, while the experimental group of vision abnormality of cynomolgus monkeys in natural white light simulation condition, the accuracy rate was 75% within 5 times, the accuracy rate was 45% within 5 times in dim 17:00-18:00 natural light simulation condition, and the food could not be effectively taken in dim 18:00-19:00 natural light simulation condition. The experimental results are consistent with the characteristics of early-onset retinal degeneration disease, and the vision is worse in the dark and the disease worsens with age.

[0039] Table 1

[0040] In-cage grouping / status M3 performance of experimental group with abnormal vision Control group WT01, WT02 Under white light / light Exploration + eye fixation on food dish + 75% accuracy within 5 attempts Completed quickly and accurately Dim 17:00-18:00 natural light Exploration + reduced eye fixation + 45% accuracy within 5 attempts Completed quickly and accurately Dim 18:00-19:00 natural light Exploration + no eye fixation + near 0 or no accuracy Completed quickly and accurately Dark after 19:00 Did not approach, did not eat Did not approach, did not eat

[0041] Example 2 Unmanned climbing motion behavior experiment of non-human primates in a single behavior device cage

[0042] 1. Experimental device and environment

[0043] A variable height climbing cage was customized, the total height of which could be set to 1500mm (the height could be adjusted according to the height characteristics of the animal) Figure 3 ), and was provided with two partitions and three independent environments of different heights. Taking the height of 1500mm as an example, the height could be adjusted to 500mm, 1000mm and 1500mm. Three sides of the cage are iron frames for non-human primates to climb, and one side is made of transparent acrylic material for easy shooting and observation. The cage is placed in a laboratory with ventilation and no disturbing noise, toys are placed in the cage, and a monitoring camera is provided for recording animal motion behavior, and a sound system is provided for playing music simulating natural environment or music familiar to animals. The companion cage is placed beside the variable height climbing cage to reduce the anxiety of experimental animals. The indoor illuminance is set to two groups of light environments of 1000 lux and above and 250 lux and below, or adjusted according to the experiment.

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

[0045] 2. Operation method

[0046] The above device is used to test the climbing behavior of primates under different light conditions. The experiment is divided into two groups of light environment, and the light intensity is measured using a light meter. The light intensity is 1000 lux and above, and 250 lux and below, which can be roughly corresponding to natural white light at noon 12:00-13:00 and natural light at dusk 18:00-19:00 in the afternoon. The time or light intensity can also be adjusted according to the experimental requirements. In each group of light environment, the animals are placed in a variable height cage. First, adjust the cage height to 500 mm. After 5 minutes of free activity without human interference, start recording for 15 minutes. Remove the adjacent partition, adjust the height to 1000 mm, and after 5 minutes of free activity without human interference, start recording for 15 minutes. Remove the adjacent partition again, adjust the height to 1500 mm, and record as above. Each height in each group of light environment is repeated more than three times. (For young animals, rapid spinning, constant screaming and other behaviors may occur 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 to the environment. After the mood stabilizes, the experiment is carried out.) Finally, analyze the recorded animal movement behavior video.

[0047] 3. Result analysis

[0048] The behavior of animals in different light environments and different cage heights is recorded, and the time taken by the animal to climb or descend the cage once, the climbing speed and frequency, and the number of jumps per minute in the cage are quantified. Normal primates often exhibit rapid climbing up and down and multiple jumps in the cage, while visually impaired animals often exhibit significantly reduced climbing speed and jump frequency due to their inability to clearly see the cage bars, and they often exhibit exploratory movements when climbing. The climbing up and down record standard is that the animal's upper limbs tightly grip the cage bars, the lower limbs are lifted to start timing, and the lower limbs are smoothly landed on the platform to end, which is recorded as once. The jump record standard is that the animal jumps from the original place to another place and tightly grips the cage bars, which is recorded as once.

[0049] Table 2 and Table 3 are the statistical tables of the climbing behavior experiment of the 1-year-old cynomolgus monkeys in the variable height climbing cage. The 1-year-old cynomolgus monkey WT01 in the control group and the 1-year-old abnormal vision cynomolgus monkey M3 in the experimental group were subjected to the variable height climbing cage climbing experiment, video recording and statistical analysis. The results showed that the control group cynomolgus monkey WT01 had the behavior characteristics of rapid rotation and high activity in the 500mm variable height climbing cage. The climbing time in the 1000mm variable height climbing cage was 1-2 seconds, and the climbing time in the 1500mm variable height climbing cage was 2-3 seconds, and the cage jumping frequency was 2-5 times / min. The abnormal vision cynomolgus monkey M3 in the experimental group had the behavior characteristics of slow rotation and sometimes squatting in the corner in the 500mm variable height climbing cage. The climbing time in the 1000mm variable height climbing cage was 3-7 seconds, and the climbing time in the 1500mm variable height climbing cage was 5-13 seconds, and there was no jumping action during the period, which was significantly different from the control group. The experimental results are consistent with the characteristics of early retinal degeneration disease.

[0050] Table 2 Statistical table of high cage climbing experiment of abnormal vision cynomolgus monkey M3 in experimental group

[0051]

[0052]

[0053] Table 3 Statistical table of high cage climbing experiment of WT01 in control group

[0054]

[0055] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application and do not limit the protection scope of the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present application.

Claims

1. A method for testing visual function in a non-human primate, characterized in that, The first experiment comprises the following steps: S1, placing the test device in front of the non-human primates in the test group so that the animals can put their hands into the test device; S2, recording the behavior of the animals in the following tests, which are tests of different light modes and randomly placing the favorite objects of the animals in the test device, recording the time from placing the objects to the animals grabbing the objects, the number of times the animals stretch their hands per unit time, the degree of eye gaze of the animals, and the influence of the color preference of the objects on the animals' grabbing; S3, using the above behavior as a test index of the visual function of non-human primates to investigate the visual function of non-human primates.

2. The method of testing visual function of a non-human primate according to claim 1, wherein, The light modes in step S2 are 1000 lux and above, 500 lux, 250 lux, 125 lux and below, respectively.

3. The method of testing visual function of a non-human primate according to claim 1, wherein, The test device in step S1 is a feeding box with multiple grids, and the material of the feeding box is acrylic. The size of each grid in the feeding box is between 1.0-3.0 cm.

4. A method for testing visual function in a non-human primate, characterized in that, The second experiment also comprises the following steps: (1) the time taken by the animals to climb up or down the cage once; (2) the climbing speed and frequency of the animals in the cage; (3) the number of jumps per minute of the animals in the cage.

5. The method of testing visual function in non-human primates according to claim 4, wherein, The light modes are 1000 lux and above, and 250 lux and below.

6. The method of testing visual function in non-human primates according to claim 5, wherein, The operation of the second experiment is to record the behavior of the animals in the test cage under the light modes of 1000 lux and above, and 250 lux and below, respectively. Specifically, the following steps are included: (1) light mode is 1000 lux and above S1, placing the non-human primates in a test cage with variable height, adjusting the cage height to the first height, recording the behavior of the animals after they freely move in the mode without human disturbance, and the recording time is 15 minutes; S2, adjusting the cage height to the second height, recording the behavior of the animals after they freely move in the mode without human disturbance, and the recording time is 15 minutes; S3, adjusting the cage height to the third height, recording the behavior of the animals after they freely move in the mode without human disturbance, and the recording time is 15 minutes; (1) light mode is 250 lux and below, repeating S1-S3.

7. The method of testing visual function of a non-human primate according to claim 6, wherein, The test cage is set to three heights, with a total height of 1500 mm, and is provided with two partitions which are inserted into the climbing cage in the form of plugs. Initially, the animals are placed in the first partition space from bottom to top, which is the first height of the test cage, 500 mm. When the first partition from bottom to top is removed, it is the second height of the test cage, 1000 mm. When the second partition from bottom to top is removed, it is the third height of the test cage, 1500 mm.

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