Mouse visual detection method and system based on visual motion response and gait analysis

Abstract

The invention discloses a mouse visual detection method and system based on visual motion response and gait analysis, and relates to the technical field of animal visual detection. The mouse visual detection method based on visual motion reaction and gait analysis comprises the following steps: arranging eight groups of spatial frequency change or contrast change rotating gratings in a visual motion detection box body, combining every two of the eight groups of spatial frequency or contrast gradients into one stimulation condition, forming four stimulation conditions, and performing visual motion detection on the four stimulation conditions; sequentially stimulating the to-be-tested mouse under four stimulation conditions; the head follow-up of the mouse is monitored in real time through the visual motion reaction system, and the real-time rotation speed of the head is obtained; the moving information of each foot of the mouse subjected to visual motion reaction detection is acquired in the active gait detection box with the red and green mixed light source, and the motion coordination is evaluated. The mouse vision-perception-movement loop can be effectively evaluated by combining the visual movement response and the gait movement.

Description

Technical Field

[0001] The present invention relates to the technical field of visual detection, and in particular to a mouse visual detection device and a detection method based on visual motion response and gait analysis. Background Art

[0002] Vision, as a core mechanism for the interaction between organisms and their surroundings, has important adaptive functions. It not only plays a key role in hunting and foraging, but also has a significant impact on social behavior, spatial navigation, and learning ability. A sensitive visual system enables organisms to quickly identify changes in the environment so that they can respond in a timely manner and enhance their chances of survival and reproduction. In addition, the processing of visual information is closely related to cognitive ability, which further enriches organisms' understanding and adaptation to the world around them. The combined effect of these functions makes vision an indispensable research topic in ecological and biological studies.

[0003] During the process of visual formation, light enters the eye through the cornea and lens and focuses on the retina. After being captured by the photoreceptors (cones and rods) in the retina, it is converted into electrical signals. These electrical signals are converted into neural signals by bipolar cells and ganglion cells and transmitted along the optic chiasm to the lateral geniculate body (LGN) of the brain. LGN is part of the thalamus and is an important relay station for visual information. LGN performs preliminary integration of visual information from both eyes. The integrated visual information is transmitted to the primary visual cortex (V1, located in the occipital lobe) of the brain through visual amplification. V1 is responsible for basic visual processing, such as edge detection and direction selection. Subsequently, the visual information processed by the primary visual cortex (V1) is further transmitted to higher-order visual areas, such as V2, V3, V4, and V5 / MT. These areas are responsible for more complex visual processing, including color recognition, motion perception, and object recognition. The information processed by the higher-order visual areas is transmitted to the parietal lobe (especially the dorsal visual pathway, the "where" pathway). The parietal cortex is involved in spatial perception and motor planning, integrating visual information and converting it into spatial and motor-related signals. Movement-related information in the parietal cortex is transmitted to the prefrontal cortex and premotor areas, which are responsible for the formulation of motor plans and strategies. Subsequently, the information is transmitted to the primary motor cortex (M1), which is the area of ​​the brain responsible for executing specific motor commands. The motor commands generated by the primary motor cortex are transmitted to the spinal cord through the pyramidal tract. Among them, the pyramidal tract is the main motor pathway between the brain and the spinal cord, and is mainly responsible for transmitting the motor commands generated by the brain to the motor neurons in the spinal cord. After receiving the commands from the pyramidal tract, the motor neurons in the spinal cord transmit these signals to the peripheral nerves and finally reach the muscles, mediating muscle contraction and movement execution. During the execution of the movement, sensory feedback (including visual feedback) is transmitted back to the brain through the sensory nerves, constantly adjusting and correcting the movement to ensure the accuracy and coordination of the movement.

[0004] In short, visual cortical signals are integrated through a series of complex neural pathways and brain regions, and are ultimately transmitted to motor nerves to achieve the integration of visual information and motor commands. This process involves the processing of visual signals in the primary visual cortex and higher-order visual areas, through the parietal cortex to the motor cortex, and then through the spinal cord to the muscles, forming a complete sensory-motor circuit, and vision determines the motor response of mice to a certain extent. This complex neural network ensures that visual information can accurately guide motor behavior. It is of great significance to deeply explore and understand the interaction between the perceptual and motor systems and the visual-perceptual-motor integration mechanism in neuroscience.

[0005] There are many methods for evaluating animal visual function. Among them, histology, as a common method for evaluating animal visual function, must sacrifice the life of the animal in order to quantify the thickness of neurons in each layer of the retina and reveal the integrity of the retinal structure. However, the integrity of the retinal structure is not completely equivalent to the soundness of visual function. In order to make up for this problem, a variety of electrophysiological detection methods have been born, such as electroretinogram (ERG) and visual evoked potential (VEP). Electrophysiological detection can effectively reflect the processing and transmission of neural signals and provide dynamic and real-time functional data, thereby reducing the influence of subjective factors on the results. However, there are many technical barriers and operational problems in electrophysiological detection, such as the extremely high professionalism and operational technical requirements of the experimenters; the detection environment is often limited to the laboratory environment, and it is impossible to truly reflect the visual performance of animals in nature; there is a risk of death due to anesthesia during the experiment, which may cause ethical disputes; during the VEP experiment, surgery is required to insert electrodes into the animals, which may cause pain and stress reactions, and is invasive. In subsequent explorations, people found that the visual-motor response test developed based on animal behavior can simultaneously combine the two major advantages of non-invasiveness and objectivity, opening up a new path for visual function assessment. The visual-motor response system often uses high-precision camera equipment to capture the head movement of mice under different stimulation conditions, and detects their ability to respond to moving or changing stimuli to evaluate visual acuity and motion perception. Although the instrument provides a rapid means of visual-motor response assessment, its visual-motor response principle based on head rotation speed mainly relies on the accessory optic nerve tract (AOT) to transmit visual signals to relatively low-level nuclei such as the medial terminal nucleus (MTN) and the optic tract nucleus (NOT), and this simple basic reflex does not involve the complex processing of the visual cortex. Therefore, there is an urgent need for a detection method to further make up for the defects of instrument inspection and fully reveal the correlation between the complete visual pathway and the motor circuit. This detection defect can be further clarified using an active gait meter.

[0006] The current detection methods are limited to the use of a single instrument. For example, the visual motor response only detects the mouse's vision, ignoring whether the mouse's motor nerves and body are healthy and complete. The use of a single active gait does not consider whether the mouse's vision is within a healthy range, which in turn affects the motor system. There are serious loopholes in its detection methods. The main defects and deficiencies include:

[0007] ①There are limitations in visual function testing:

[0008] 1. Commonly used visual detection methods, such as the optokinetic response test, cannot eliminate the influence of the mouse's own limb coordination and health status on the error of vision parameters.

[0009] 2. It is unknown whether the visual feedback to motor response circuit is within a normal controllable range.

[0010] ②Motor function testing has limitations:

[0011] After excluding differences in mouse weight and body shape, whether the abnormal data obtained during movement detection is related to the visual status of the mice. Summary of the invention

[0012] In view of the deficiencies in the prior art, the present invention provides a mouse vision detection device and a detection method based on visual-motor response and gait analysis, which solves the problem of limitations in visual function detection caused by the use of visual-motor response alone in the prior art.

[0013] To achieve the above objectives, the present invention is implemented by the following technical scheme: a mouse vision detection method based on visual motion response and gait analysis, comprising the following steps:

[0014] Step 1: Eight groups of rotating gratings with changing spatial frequencies or contrast gradients are set in the optokinetic detection box, and the eight groups of spatial frequencies or contrast gradients are combined into one stimulation condition in pairs, forming four stimulation conditions, and the four stimulation conditions are used to stimulate the mice in the optokinetic detection box in turn;

[0015] Step 2: Use a high-speed camera to shoot the mice in the visual motion detection box to obtain the real-time head rotation speed of the animals, which is used to calculate the visual motion reaction time difference and the visual motion reaction index;

[0016] Step 3: Repeat the stimulation of the eight groups of rotating grating conditions for 12 times on the mice in the optokinetic detection box, and generate the head rotation speed histogram and optokinetic response index heat map through the 12 stimulations;

[0017] Step 4: After the visual movement test, the walking index of the mice is measured in an active gait measurement instrument;

[0018] Step 5: According to the quadruped information of the mouse recorded by active gait, analyze and obtain the walking time, walking cycle, number of steps per unit time, standing time and foot support ratio of the mouse;

[0019] Step 6: Compare the vision of different strains of mice by obtaining the visual-motor reaction time difference, visual-motor reaction index and active gait data.

[0020] Furthermore, the spatial frequencies presented by the rotating gratings of eight groups of spatial frequency gradients are: 0.02cyc / °-0.1cyc / °; 0.15cyc / °-0.2cyc / °; 0.25cyc / °-0.3cyc / °; 0.4cyc / °-0cyc / °; the contrast gradients presented by the rotating gratings of eight groups of contrast gradients are 100%-80%; 50%-25%; 12.5%-10%; 5%-0%.

[0021] Furthermore, the rotating grating has a rotation speed of 12° / s.

[0022] Furthermore, the single stimulation condition repeatedly stimulates the mouse in the visual motion detection box 12 times, with each stimulation lasting 50 seconds and an interval between stimulations of 5 seconds.

[0023] Furthermore, the visual-motor reaction time difference ΔT = T 正 -T 反 ; Optomotor response index

[0024] Furthermore, the head rotation speed in the visuomotor response index heat map is in the range of 0-24° / s; the head rotation speed histogram is a normalized head rotation speed histogram between -60° / s and 60° / s.

[0025] Furthermore, the active gait measuring instrument has an ambient light source, which includes an active gait green light source and an active gait red light source; the active gait green light source is controlled within the range of 3-50l ux.

[0026] On the other hand, the present invention also provides a mouse vision detection device based on visual-motor response and gait analysis, comprising: a visual-motor detection box, which serves as a place for preliminary detection of the visual sensitivity of the mouse; and an active gait measuring instrument, which is used to detect the movement coordination of the mouse.

[0027] Furthermore, the visual motion detection box includes four display screens forming a frame-shaped structure and a plane mirror arranged on the lower side of the box body on the frame structure, wherein the display screen is used to provide virtual rotating grating stimulation; a high-speed camera for tracking the movement of the mouse head is arranged in the central area of ​​the upper plane mirror; and a movable circular platform can be placed on the plane mirror on the lower side of the box body.

[0028] Furthermore, the active gait measuring instrument includes an active gait detection box, which is a tempered glass box with an open top, and the bottom of the tempered glass box is an experimental runway; a high-speed camera 2 is provided below the central area of ​​the tempered glass box, a red light source is provided at the top of the tempered glass box, and a green light source is provided at the bottom of the tempered glass box.

[0029] The present invention has the following beneficial effects:

[0030] (1) The operation method has low professional requirements for experimental personnel and is easy to use, suitable for researchers with different experience levels.

[0031] (2) Non-invasive and direct vision detection method.

[0032] (3) It can directly demonstrate the connection between visual ability, perception and motor circuits quickly and efficiently.

[0033] (4) Add new detection indicators to supplement the visual detection errors caused by a single detection instrument.

[0034] In short, the combined visual-motor response and gait movement can effectively evaluate the visual-perception-motor circuit of mice. On the basis of visual function evaluation, it further expands the detection of motor function-related indicators based on vision guidance, which helps to reveal the complex interactive relationship between vision and movement. Through multi-dimensional comprehensive evaluation, the present invention can more accurately identify abnormal patterns of animal behavior, providing strong support for disease diagnosis, efficacy evaluation and rehabilitation research. On the basis of ensuring ethics, the operation process is simple and easy to understand, which brings unprecedented convenience to scientific researchers.

[0035] Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 It is a flow chart of the mouse vision detection method based on visual motion response and gait analysis of the present invention;

[0037] Figure 2 The rotating grating images of different spatial frequency gradients of the present invention;

[0038] Figure 3 The wild type and Pde6b under different spatial frequency gradient stimulation of the present invention rd10 Change curves of ΔT and OMR of mice;

[0039] Figure 4 The wild type and Pde6b under different spatial frequency gradient stimulation of the present invention rd10 Histogram of head rotation speed of mice;

[0040] Figure 5 The wild type and Pde6b under different spatial frequency gradient stimulation of the present invention rd10 Heat map of the optokinetic response of mice;

[0041] Figure 6 The rotated grating images of different contrast gradients of the present invention;

[0042] Figure 7 The wild type and Pde6b under different contrast gradient stimulation of the present invention rd10 The curves of ΔT and OMR changes in mice;

[0043] Figure 8 The wild type and Pde6b under different contrast gradient stimulation of the present invention rd10 Head rotation speed histograms of ΔT and OMR of mice;

[0044] Fig. 9 The wild type and Pde6b under different contrast gradient stimulation of the present invention rd10 Heat map of the optokinetic response of mice;

[0045] Fig.10 The wild type and Pde6b of the present invention after the visual motion reaction test rd10 Walking pattern diagram;

[0046] Fig.11 The wild type and Pde6b of the present invention after the visual motion reaction test rd10 of marching charts, footprints and foot support cases;

[0047] Fig.12 The wild type and Pde6b of the present invention after the visual motion reaction test rd10 Through the statistical charts of walking time, walking cycle, number of steps per unit time, and standing time of fixed detection length;

[0048] Fig.13 The wild type and Pde6b of the present invention after the visual motion reaction test rd10 Statistical parameters of foot support phase percentage;

[0049] Fig.14 It is a structural schematic diagram of the visual motion detection box of the present invention;

[0050] Fig.15 It is a structural schematic diagram of the movable platform in the present invention;

[0051] Fig.16 Schematic diagram of the structure of the active gait box.

[0052] In the figure, 110, plane mirror; 120, display screen; 130, high-speed camera 1; 140, movable platform; 210, tempered glass box; 220, ambient light source; 230, high-speed camera 2. DETAILED DESCRIPTION

[0053] According to the following Figure 1-Figure 16 The present invention provides a method and system for visual detection of mice based on visual motion response and gait analysis.

[0054] like Figure 1 On the one hand, an embodiment of the present invention provides a mouse vision detection method based on visual motion response and gait analysis.

[0055] The following steps are involved:

[0056] Step 1: Eight groups of rotating gratings with varying spatial frequencies or contrast gradients are set in the optokinetic detection box. The eight groups of rotating gratings are combined in pairs into one stimulation condition, forming four stimulation conditions. The four stimulation conditions are used to stimulate the mice in the optokinetic detection box in turn.

[0057] Step 2: Use a high-speed camera 130 to shoot the mice in the visual motion detection box to obtain the real-time head rotation speed, which is used to calculate the visual motion reaction time difference and the visual motion reaction index;

[0058] Step 3: Eight groups of rotating gratings are used to repeatedly stimulate the mouse in the optokinetic detection box 12 times, and a head rotation speed histogram and an optokinetic response index heat map are generated through the 12 stimulations;

[0059] Step 4: After the visual movement test, the walking index of the mice is measured in an active gait measurement instrument;

[0060] Step 5: According to the quadruped information of the mouse recorded by active gait, analyze and obtain the walking time, walking cycle, number of steps per unit time, standing time and foot support ratio of the mouse;

[0061] Step 6: Compare the vision of different strains of mice by obtaining the visual-motor reaction time difference, visual-motor reaction index and active gait data.

[0062] In the above embodiment, it should be noted that the two-month-old wild-type (normal vision mice) and Pde6b rd10 Mice (classic retinitis pigmentosa model mice, blind mice whose vision is almost completely degenerated at 2 months of age) were respectively obtained using the visual motion response system and active gait measurement instrument. Before the visual motion response test, the experimental mice needed to be placed in the experimental room one day in advance and protected from light with black light-shielding cloth to adapt to the experimental environment.

[0063] A single stimulation condition repeatedly stimulates the mice in the visual motion detection box 12 times (it should be noted that the eight groups of rotating gratings are fixed in pairs. For example, if the stimulation condition is 0.02cyc / °-0.1cyc / °, then 0.02cyc / ° and 0.1cyc / ° are measured together, each repeated 12 times, 12 times 0.02cyc / ° and 12 times 0.1cyc / °, that is, the number of repetitions in the four stimulation conditions is 24 times, and the 24 repetitions in the combination are randomly arranged). Each stimulation time is 50s, and the interval between stimulations is 5s. After each group of stimulations, the mice can rest for 1-2 hours. During the test, it is necessary to pay attention to the status of the mice at all times. If the mice jump off the experimental platform too frequently, they need to be put back in the cage to rest for 10-15 minutes before recording the experiment.

[0064] The visual motion response system records the head rotation speed of mice in real time through a high-speed camera-130. According to the grating rotation speed of 12° / s, the head rotation speed of mice in the range of 2-14° / s is defined as the following behavior induced by the stimulus, and other head rotation speeds are judged as invalid following; the direction of head rotation consistent with the grating rotation is called positive following, and the opposite direction is called reverse following; according to the grating rotation direction, it can be further subdivided into clockwise positive following, clockwise reverse following, counterclockwise positive following, and counterclockwise reverse following. Here, the time difference (ΔT) and ratio (OMR value) of positive following and reverse following in the clockwise and counterclockwise directions are quantified.

[0065] Preferably, the head rotation speed in the visual-motor response index heat map is within the range of 0-24° / s; however, it should be noted that each point in the visual-motor response index heat map represents the OMR value of the head rotation speed within the range of [0 / 12]-[12 / 24]° / s, and actually the horizontal axis represents the rotation speed range of 0-12° / s; the vertical axis represents the rotation speed range of 12-24° / s, and each point corresponds to a range between the horizontal and vertical axes, indicating that the point is an OMR value greater than the horizontal axis and less than the range of the vertical axis.

[0066] The head rotation speed histogram is a normalized head rotation speed histogram between -60° / s and 60° / s.

[0067] By analyzing the average results of 12 repetitions for each stimulus, the corresponding head rotation speed histogram and optokinetic response heat map were generated to more intuitively represent the visual acuity situation.

[0068] Optionally, the eight groups of rotating gratings are eight groups of rotating gratings with spatial frequency gradients; or eight groups of rotating gratings with contrast gradients; when eight groups of rotating gratings with spatial frequency gradients are used, the rotating gratings present a series of changes in spatial frequency of 0-0.4cyc / °; when eight groups of rotating gratings with contrast gradients are used, the rotating gratings present a series of changes in contrast gradient of 0% to 100%.

[0069] Above: Visual reaction time difference ΔT = T 正 -T 反 , and the average value of ΔT obtained from 12 stimulations was taken to draw a curve graph;

[0070] Optomotor Response Index The average value of OMR obtained from 12 stimulations was taken to draw a curve graph.

[0071] In this embodiment, two types of rotating gratings with gradient changes are provided, one of which is a rotating grating with eight groups of spatial frequency gradient changes, and the other is a rotating grating with eight groups of contrast gradients.

[0072] In this embodiment, the experimental parameters of the corresponding rotating grating should be set before the experiment.

[0073] 1) When using eight sets of rotating gratings with spatial frequency gradients, the grating changes are as follows: Figure 2 As shown, the grating fringes become finer as the spatial frequency increases.

[0074] like Figure 3 As shown, in this embodiment, the eight groups of spatial frequency gradients are 0cyc / °, 0.02cyc / °, 0.10cyc / °, 0.15cyc / °, 0.2cyc / °, 0.25cyc / °, 0.3cyc / ° and 0.4cyc / °, which are combined in pairs to form four groups of spatial frequency stimulation conditions, namely: 0.02cyc / °-0.1cyc / °; 0.15cyc / °-0.2cyc / °; 0.25cyc / °-0.3cyc / °; 0.4cyc / °-0cyc / °. Figure 3 a and Figure 3 In b, the wild-type mice showed a trend of first rising and then falling with the increase of spatial frequency, and the peak usually occurred at 0.15cyc / ° or 0.2cyc / °; at this time, the average visual motion reaction time difference ΔT was 8.725±0.875s, and the visual motion reaction index OMR value was 2.276±0.155s.

[0075] The head rotation speed is normalized according to the frame ratio, and the head rotation speed histogram ( Figure 4 ), it can be seen that when the positive follow-up (yellow bar) is returned to 1, the reverse follow-up (blue bar) shows an inverted "U" trend. Figure 5 ), the area of ​​the red region (OMR value>2) first increases and then decreases with the increase of spatial frequency.

[0076] 2) When using eight sets of contrast gradient rotating grating experiments, the grating changes are as follows Figure 6 As shown, as the contrast decreases, the difference between black and white becomes smaller and tends to be consistent.

[0077] like Figure 7 In this embodiment, the eight sets of contrast gradient rotating gratings are 0%, 5%, 10%, 12.5%, 25%, 50%, 80% and 100%, respectively. After two sets of contrast gradient stimulation conditions, four sets of contrast gradient stimulation conditions are formed, namely 100% to 80%; 50% to 25%; 12.5% ​​to 10%; 5% to 0%. As the contrast gradient decreases, the difference in visual motion reaction time ΔT and the visual motion reaction index OMR value of wild-type mice gradually decrease from 7.915±0.520s at 100% contrast gradient to -0.153±0.443 at 0% contrast gradient. The visual motion reaction index heat map ( Fig. 9 ) also gradually changed from red (OMR value > 2) to a large area of ​​blue (OMR value < 1.2), and the head rotation speed histogram ( Figure 8 ), it can be seen that when the positive following motion (yellow bar) is returned to 1, the negative following motion (blue bar) shows a gradually decreasing trend.

[0078] However, Pde6b rd10 The visual motion response of mice did not change with the change of spatial frequency gradient or contrast gradient. When the spatial frequency was 0.2cyc / °, the ΔT and OMR values ​​were 0.601±0.436s and 1.145±0.076, respectively; the lowest values ​​of ΔT and OMR appeared at 0cyc / °, which were -0.842±0.553s and 0.956±0.061, respectively. Under different contrast gradients, the maximum ΔT was 0.844±0.575s and the minimum was -0.367±0.584s; the maximum and minimum OMR values ​​were 0.140±0.066 and 1.000±0.059, respectively. The data showed that blind mice could not respond to environmental stimuli and make corresponding visual following responses.

[0079] In summary, the visual motion response has a certain degree of high sensitivity and real-time performance in detecting the visual function of mice. It can accurately capture the gaze movement and response of mice and provide rich data on visual attention and perception. However, the visual motion response has strict requirements on environmental lighting and the adaptability of mice, which may affect the repeatability of the data to a certain extent. In addition, some mouse strains or disease models may not cooperate well with visual motion detection, such as blind mice, movement disorders and other mouse models. Therefore, it is necessary to combine other instruments to make up for this detection loophole.

[0080] Therefore, the active gait detection box in step 4 is used here for detection, and the walking index is obtained through the active gait measurement instrument.

[0081] After the visual motion reaction test, the mice need to be placed in the experimental room one day in advance and protected from light with black light-shielding cloth to adapt to the experimental environment before active gait detection can be performed. Before the experiment, set the corresponding experimental parameters. Each mouse was tested at least 30 times. After screening, the valid data retained should be no less than 6 times to ensure the reliability of the data.

[0082] Preferably, the active gait measuring instrument has an ambient light source 220, and the ambient light source 220 includes an active gait green light source and an active gait red light source.

[0083] Among them, the active gait green light source is controlled within the range of 3-50l ux. The green light belongs to the side light footprint enhancement device, which is helpful for identifying footprints; the active gait red light source is controlled within the range of 3-50l ux. The red light is the top background light, which can increase the contrast of footprints.

[0084] Active gait clearly demonstrates wild-type mice and Pde6b rd10 Differences in the way mice walk. Fig.10 As shown, the foot parts are LF (red, left front foot), LH (purple, left hind foot), RF (yellow, right front foot), and RH (green, right hind foot). Fig.10 In the figure, Step Cycle is the walking cycle, Stand is the standing state, Swing is the moving distance, Initial Dual Stance is the initial double foot support, and Terminal Dual Stance is the terminal double foot support.

[0085] like Fig.11 As shown in the experiment, it was found that the walking graph of wild-type mice showed obvious regularity and consistency. The changes in their gait cycle, step frequency, etc. were relatively stable, forming a clear walking pattern. This regularity shows that wild-type mice can effectively coordinate their movements when walking and maintain good balance and sense of direction. In contrast, Pde6b rd10 Graphs of the mice's walking showed that each foot moved in an unpredictable pattern.

[0086] like Fig.12 As shown in Figure 2, further refinement of gait-related indicators reveals:

[0087] 1. If Fig.12 a, Wild-type mice and Pde6b rd10The walking time through the fixed detection length was 1.2318±0.0594 sec and 2.0089±0.1589 sec, respectively.

[0088] 2. If Fig.12 b, The step cycle of wild-type mice calculated by combining footprints is 0.2639±0.0258 seconds / cycle, while Pde6b rd10 For mice, it was 0.3651±0.0439 seconds / cycle.

[0089] 3. If Fig.12 c, the corresponding cadences are 16.1175±1.5534 steps / second and 11.6027±1.1787 steps / second, respectively.

[0090] 4. If Fig.12 d, the average single-leg standing time was 0.1424±0.0229 seconds and 0.2343±0.0394 seconds respectively.

[0091] like Fig.13 , the mice were divided into zero support (Zero), single foot support (Single), contralateral foot support (Diagonal), homologous foot support (Lateral), three-legged support (Three), four-legged support (Four) and shoulder strap support (Gridle), and Supporting Sprcies Ratio was the proportion of foot support. The data showed that the proportion of three-legged support and four-legged support in wild-type mice was Pde6b. rd10 The number of mice was significantly reduced.

[0092] It should be noted that the shoulder girdle support (Gridle) is a form of uncoordinated foot support in mice.

[0093] These data show that the gait cycle and stride length of congenitally blind mice vary greatly, and the fluctuation of step frequency is also unstable, reflecting difficulties in spatial positioning and movement coordination. The proportion of three-legged support and four-legged support may indirectly reflect the visual function of mice. The lack of vision has a significant impact on their motor ability, resulting in confusion and incoordination of walking behavior.

[0094] In summary, the wild type and Pde6b rd10 A comparative experiment was conducted on mice to test the feasibility of visual motion response and active gait measurement instrument linkage detection of vision. rd10As a common transgenic model, mice are mainly used to study retinal degenerative diseases related to retinitis pigmentosa. This mouse carries a specific gene mutation that causes defects in the development and function of the visual system. Compared with wild-type mice, Pde6b rd10 Soon after birth, mice begin to show degeneration and death of retinal photoreceptor cells (cones and rods), which is manifested by a significant decrease in vision. This retinal degeneration is progressive, and pathological changes can be observed at different stages. rd10 Mice serve as a key experimental model for the present invention.

[0095] The results of the visual motion response data showed that the ΔT and OMR values ​​of wild-type mice showed a trend of first increasing and then decreasing with the increase of spatial frequency, reaching a peak at 0.15cyc / ° or 0.2cyc / °; at the same time, these indicators steadily increased with the increase of contrast until reaching the maximum value at 100% contrast. rd10 Mice, whose vision and visual ability have been shown to be significantly impaired. The number of photoreceptors (especially rods) in their retina is reduced, resulting in reduced visual sensitivity and difficulty in effectively locating objects in low-light environments. In the visual motion response test, their response to motion stimuli is weakened, and they have no significant response to changes in the spatial frequency or contrast of the grating. Their speed and accuracy are significantly lower than those of normal mice, and their OMR values ​​are stable around 1, and their ΔT values ​​are close to zero. In addition, Pde6b rd10 Mice also have problems with spatial perception and show a higher tendency to get lost. Due to decreased visual ability, the brain may undergo a neuroremodeling reaction during the pathological process, affecting limb coordination. Therefore, it is of great significance to explore how vision guides movement.

[0096] Active gait has obvious advantages in visually guided motion detection. Individuals receive visual information about the surrounding environment in real time, thereby flexibly adjusting their stride, speed and direction, improving walking safety and efficiency, and optimizing movement patterns. In terms of gait analysis, we have deeply explored a number of key parameters, including walking time, walking cycle, step frequency, standing time, percentage of support phase, etc. These parameters are different between wild-type mice and Pde6b mice. rd10 There was a significant statistical difference between the blind mice and the wild-type mice. In the active gait measurement instrument, the time to pass the unit detection length ranged from about 1.1724 to 1.2912 seconds. However, Pde6b rd10 The passage time of mice was mainly concentrated between 1.8500 and 2.1678 seconds. rd10 The mice had significantly longer ground contact time than wild-type mice, and the walking cycle in the walking graph was chaotic. In particular, the percentage of tripod and quadruped support phases showed a significant upward trend.

[0097] In summary, mice with better vision pass through the unit detection range faster, and the proportion of three-legged and four-legged support is lower than that of mice with poor vision, and the number of steps per unit length is less. The data shows that mice with good vision have better motor coordination.

[0098] The present invention compares wild-type mice and Pde6b rd10 The study revealed the important influence of visual ability on motor performance. Wild-type mice showed good visual response and flexible gait in the visual motor response test, while Pde6b rd10 Due to the reduction in the number of photoreceptors, the visual ability of mice is significantly impaired, resulting in poor coordination, prolonged walking time, and changes in the percentage of support phase during movement. This study emphasizes the key role of visual information in guiding movement and spatial perception, and shows that visual defects can significantly reduce movement efficiency. This combined detection method not only enriches the dimensions of visual function evaluation, but also directly demonstrates the relationship between vision and motor function for the first time, providing an important basis for a deeper understanding of the impact of visual impairment on animal behavior patterns. Compared with existing behavioral comparisons for visual assessment, it has the following advantages:

[0099] 1. The operation method has low professional requirements for experimenters, is easy to use, and is suitable for researchers of different levels to implement.

[0100] 2. Non-invasive and direct visual detection method.

[0101] 3. It can directly demonstrate the connection between visual ability, perception and motor circuits quickly and efficiently.

[0102] 4. Add new detection indicators to supplement the visual detection errors caused by a single detection instrument.

[0103] In short, the combined visual-motor response and gait movement can effectively evaluate the visual-perception-motor circuit of mice. On the basis of visual function evaluation, it further expands the detection of motor function-related indicators based on vision guidance, which helps to reveal the complex interactive relationship between vision and movement. Through multi-dimensional comprehensive evaluation, the present invention can more accurately identify abnormal patterns of animal behavior, providing strong support for disease diagnosis, efficacy evaluation and rehabilitation research. In addition, the data summarized in this patent can be combined with AI algorithms to realize the automation and efficiency of the evaluation process, greatly improving the accuracy of the data obtained. On the basis of ensuring ethics, the operation process is simple and easy to understand, which brings unprecedented convenience to scientific researchers.

[0104] Quantifying the time difference (ΔT) and ratio (OMR value) of the visual-motor response can reflect the visual function of mice under different spatial frequencies or contrast conditions. Measuring the walking time, walking cycle, step frequency and foot support ratio of mice through active gait can reveal the perception-motor circuit of mice. By combining the two instruments and comparing the obtained data, the defects of single instrument detection can be compensated and the visual-perception-motor circuit can be directly revealed.

[0105] It should be noted that Pde6b rd10 The Pde6b mouse is a transgenic model used primarily to study retinal degenerative diseases related to retinitis pigmentosa. The mouse carries a specific gene mutation that causes defects in the development and function of the visual system. Compared with wild-type mice, Pde6b rd10 Soon after birth, mice begin to show degeneration and death of retinal photoreceptor cells (cones and rods), which is manifested by a significant decrease in vision. This retinal degeneration is progressive, and pathological changes can be observed at different stages. rd10 Mice serve as a key experimental model for the present invention.

[0106] On the other hand, the present invention also provides a mouse vision detection device based on visual motion response and gait analysis, such as Figure 14-16 As shown, the mouse vision detection device based on visual-motor response and gait analysis includes a visual-motor detection box and an active gait detection box. The visual-motor detection box is used as a place for preliminary detection of the visual sensitivity of mice; the active gait detection box is used as a place for evaluating the movement coordination of mice.

[0107] The visual motion detection box includes four display screens 120 formed into a frame structure and plane mirrors 110 arranged on the upper and lower sides of the frame structure. The display screen 120 is used to provide a rotating grating. The central area of ​​the top plane mirror 110 is provided with a high-speed camera 130 for shooting images in the visual motion detection box; the upper surface of the lower tempered glass 110 is provided with a movable platform 140.

[0108] Preferably, the length, width and height of the visual motion reaction box are respectively: 66 cm×66 cm×40 cm. It is worth noting that one of the display screens 120 is made into an openable door structure to facilitate placing mice in or taking them out of the visual motion detection box.

[0109] Preferably, the movable platform 140 is made of polymethyl methacrylate, with a height of 13 cm and a diameter of 5.2 cm.

[0110] Preferably, the average brightness of the white grating in the visual motion display screen 120 is 10-80 cd / m 2 The average brightness of the black grating in the visual motion response display screen 120 is 0.1-0.5 cd / m 2 within the range.

[0111] like Fig.16 The above-mentioned active gait measuring instrument includes an active gait detection box, which is formed by a tempered glass box 210 with an open top. The tempered glass box 210 is an experimental runway, and the mice move in the experimental runway.

[0112] Preferably, the width of the test track is 5.5 cm and the length of the test track is 28 cm.

[0113] In addition, a high-speed camera 230 is provided in the lower center area of ​​the tempered glass box 210, an active gait red light source is provided in the upper center area of ​​the tempered glass box 210, and an active gait green light source is provided on the side.

[0114] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0115] The preferred embodiments of the present invention disclosed above are only used to help illustrate the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to the specific implementation methods described. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A mouse vision detection method based on visual motion response and gait analysis, comprising the following steps: Step 1: Eight groups of rotating gratings with changing spatial frequencies or contrast gradients are set in the optokinetic detection box, and the eight groups of spatial frequencies or contrast gradients are combined into one stimulation condition in pairs, forming four stimulation conditions, and the four stimulation conditions are used to stimulate the mice in the optokinetic detection box in turn; Step 2: Use a high-speed camera to shoot the mice in the visual motion detection box to obtain the real-time head rotation speed of the animals, which is used to calculate the visual motion reaction time difference and the visual motion reaction index; Step 3: Repeat the stimulation of the eight groups of rotating grating conditions on the mice in the optokinetic detection box 12 times each, and generate the head rotation speed histogram and optokinetic response index heat map through the 12 stimulations; Step 4: After the visual movement test, the walking index of the mice is measured in an active gait measurement instrument; Step 5: According to the quadruped information of the mouse recorded by active gait, analyze and obtain the walking time, walking cycle, number of steps per unit time, standing time and foot support ratio of the mouse; Step 6: Compare the vision of different strains of mice by obtaining the visual-motor reaction time difference, visual-motor reaction index and active gait data.

2. A method for detecting mouse vision based on visual motion response and gait analysis according to claim 1, characterized in that: The spatial frequencies of the rotating gratings of eight groups of spatial frequency gradients are: 0.02cyc / °-0.1cyc / °; 0.15cyc / °-0.2cyc / °; 0.25cyc / °-0.3cyc / °; 0.4cyc / °-0cyc / °; The contrast gradients presented by the rotating gratings of the eight sets of contrast gradients are 100%-80%; 50%-25%; 12.5%-10%; 5%-0% in sequence.

3. A mouse vision detection method based on visual motion response and gait analysis according to claim 1 or 2, characterized in that: The rotating grating has a rotation speed of 12° / s.

4. A method for detecting mouse vision based on visual motion response and gait analysis according to claim 3, characterized in that: The single stimulation condition repeatedly stimulates the mouse in the visual motion detection box 12 times, with each stimulation lasting 50 seconds and an interval between stimulations of 5 seconds.

5. A mouse vision detection method based on visual motion response and gait analysis according to claim 4, characterized in that: Visual reaction time difference ΔT = T 正 -T 反 ; Optomotor Response Index 6. A mouse vision detection method based on visual motion response and gait analysis according to claim 5, characterized in that: The head rotation speed in the optokinetic response index heat map ranged from 0 to 24° / s; The head rotation speed histogram is a normalized head rotation speed histogram between -60° / s and 60° / s.

7. A mouse vision detection method based on visual motion response and gait analysis according to any one of claims 4 to 6, characterized in that: The active gait measuring instrument has an ambient light source, which includes an active gait green light source and an active gait red light source; Active gait green light source is controlled within the range of 3-50lux; The active gait red light source is controlled within the range of 3-50lux.

8. A mouse vision detection device based on visual motion response and gait analysis, characterized in that: include: A visual motion detection box, which is used as a place for preliminary detection of visual sensitivity of mice; An active gait meter is provided, wherein the active gait meter is used to detect the motor coordination of mice.

9. A mouse vision detection device based on visual motion response and gait analysis according to claim 8, characterized in that: The visual motion detection box includes four display screens forming a frame-shaped structure and a plane mirror arranged on the lower side of the upper box of the frame-shaped structure, and the display screen is used to provide virtual rotating grating stimulation; A high-speed camera is installed in the center of the upper plane mirror to track the movement of the mouse's head. A movable circular platform can be placed on the plane mirror at the lower side of the box.

10. A mouse vision detection device based on visual motion response and gait analysis according to claim 8 or 9, characterized in that: The active gait measuring instrument comprises an active gait detection box, which is a tempered glass box with an open top, and the bottom of the tempered glass box is an experimental runway; A high-speed camera 2 is arranged below the central area of ​​the tempered glass box, a red light source is arranged at the top of the tempered glass box, and a green light source is arranged at the bottom of the tempered glass box.

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