Non-therapeutic-purpose auditory perception training device and method for noninvasive free mouse training stimulation auditory cortex plasticity and application

Abstract

The invention discloses a non-therapeutic-purpose hearing and perception training device for non-invasive free mouse training and auditory cortex plasticity stimulation. The device comprises a soundproof box, a training device body in the soundproof box and a processing control system located outside the soundproof box. The training device main body in the soundproof box comprises a training box body, a water and air supply device, an animal action sensing device, an animal position sensing device, a loudspeaker and an illuminating lamp; the training box body comprises an open box and a dark box, and the animal position sensing device is arranged at the relative position of the junction of the open box and the dark box and the relative position of the side wall of the dark box; the processing control system located outside the soundproof box comprises a water and gas control device, a power amplifier and a control computer. The invention further discloses a method and application of the training device in auditory cortex plasticity induction and auditory perception training, and the training device has wide application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of life science training devices, and relates to an auditory perception training device, method and application for non-invasive free mouse training to stimulate the plasticity of auditory cortex for non-therapeutic purposes. Background Art

[0002] The entire developmental process of the brain is shaped by endogenous genetic factors and exogenous sensory experience input. During a specific time window of early development, the brain has extremely strong plasticity, which is called the critical period (CP). [1] At this stage, the structure and function of the brain are shaped by sensory experiences and are easily disturbed by the external environment. [2-3] The input of external sensory experience during the critical period is crucial for the development of abilities such as language. However, unlike the early developing brain, the brain circuits of adult animals are more stable, and the process of acquiring new abilities is more difficult. However, this transition from high plasticity in the early stage to a stable state in adulthood is beneficial to the individual because it ensures the consolidation and maintenance of new and more complex perceptual, motor and cognitive functions. [4-5] For example, a child's ability to use his fingers precedes his acquisition of language skills, which in turn forms the basis for the development of other functions. [6-7] However, the formation of a robust brain network in adult animals can limit future corrections. If sensory input is abnormal during early development, for example, if hearing or vision is deprived during early development, the brain may not process sensory input in the same way it was originally wired. [8] .

[0003] Studies have shown that sound exposure during the critical period can induce structural reorganization and overexpression in the auditory cortex of animals' brains related to the characteristics of the sound stimulation. For the adult auditory system, specific sound stimulation can only induce plastic changes in the auditory cortex when combined with the animal's behavior and through a period of training. [9] This paradigm in which the brain's processing of input information is altered by training and ultimately affects sensory cognitive functions is called perceptual training. If a person undergoes an auditory discrimination task to identify a specific pure tone from a group of similar sound stimuli, the average improvement factor of the subject's discrimination threshold after training is 2.4.

[10] Gradually increasing task difficulty during perceptual training has become a useful tool for assessing sensory perception. Studies of these sensory discrimination tasks in humans and animal models have provided ample evidence for rich behavioral and neural changes following training.

[11] .

[0004] Studies have shown that after frequency perception training, only the cortical area of ​​the rat primary auditory cortex neurons with task-related frequencies is strengthened.

[12] , while cortical representation areas outside of task-related frequencies are not strengthened. For this reason, the current frequency perception training paradigm can only strengthen the task-related frequency representation area in a limited way, which is manifested in the expansion of the area of ​​the task-related representation area and the strengthening of the function of the frequency representation area, while the frequency selectivity of neurons outside of the task-related representation area changes little or does not change. In addition, the current paradigm used for auditory perception training in mice often uses a fixation method after exposing the skull and then fixing it with a fixation plate. This process requires peeling off the scalp and gluing it to the skull, which is traumatic. Summary of the Invention

[0005] In order to address the deficiencies in the prior art, the purpose of the present invention is to realize a non-therapeutic and non-invasive auditory perception training method for free mice to stimulate the plasticity of the auditory cortex, and to design a matching auditory perception training device. The method and device can widely induce auditory cortical plasticity.

[0006] The auditory perception treatment method of the present invention is a non-invasive, free mouse training method for non-therapeutic purposes. It does not apply negative or painful stimulation to mice and does not rely on pain or fear to complete the required actions. The method of the present invention is applicable to a wide range of mouse training fields.

[0007] Compared with the previous method of anesthetizing experimental animals, surgically exposing the skull, gluing a restraint plate, and then fixing the animals in a holder for cognitive training, this method requires the animals to undergo anesthesia and surgery, with an incision on the head, and they need to be fixed throughout the training process, and can only move by licking with their tongues; and during the training process, due to the resistance of the animals after being fixed, the fixed restraint plate may fall off, and the animals need to be anesthetized again and surgically installed; the present invention improves on the previous training method and allows animals to train freely on the runway, providing a non-invasive and free cognitive training method;

[0008] In order to give the animals enough motivation to complete the training tasks, the animals' daily water intake needs to be adjusted before training. Each mouse needs to be deprived of water for 3 days before training, provided that the weight of the mice does not drop significantly. After each day of training, the amount of water consumed is calculated based on the number of times the animals perform correct behaviors. Each mouse should drink no more than 1 ml of water per day, and any water shortage should be accurately replenished after training. At the same time, it is also necessary to ensure that the mice remain slightly dehydrated during training, so that they have more desire to train.

[0009] The present invention provides an auditory perception training device for non-invasive, non-therapeutic training of free mice to stimulate the plasticity of the auditory cortex. The device comprises: a soundproof box and a training device body inside the box, and a processing control system located outside the soundproof box;

[0010] The training device body inside the soundproof box includes: a training box body, a water and air supply device, an animal motion sensing device, an animal position sensing device, a speaker, and a lighting lamp; the training box body includes a light box and a dark box, and the animal position sensing device is installed at the relative position of the side wall at the junction of the light box and the dark box, as well as the relative position of the side wall of the dark box;

[0011] The processing and control system located outside the soundproof box includes: a water and air control device, a power amplifier and a control computer. The control computer is configured with a MATLAB program, and controls the water and air supply device, the animal motion sensing device and the animal position sensing device through an NI data acquisition card and relays.

[0012] The training box is specifically a linear runway training box, which is made of organic glass and has dimensions of 35 cm in length, 8 cm in width, and 25 cm in height. It includes a white, opaque, uncapped light box portion (25 cm in length, 8 cm in width, and 25 cm in height) and a black, capped dark box portion (10 cm in length, 8 cm in width, and 25 cm in height). The two are directly connected to each other without obstruction, and the dark box is provided with a movable rear door to facilitate the entry and exit of experimental animals.

[0013] The water and air supply device and the animal motion sensing device are inserted into the light box from the holes on the outside of the soundproof box in parallel, with the specific insertion position being opposite to the dark box, at the end of the light box where the sealing plate is located;

[0014] Because experimental animals are timid, easily startled, and prefer dark environments, a dark box with a capped top is set up in the linear runway to provide a relatively hidden and dark environment, making the animals more inclined to stay there. The dark box serves as the starting point of the training trial. After the animals wait in the dark box for a specified time, the trial will automatically begin and a sound will be played. The experimental animals then need to exit the dark box and lick the water spout located in the water supply device in the light box. At the end of the trial, the animals need to return to the dark box to wait for the start of the next trial.

[0015] The water and air supply device includes a water supply device and an air supply device, and the water and air control device includes a water control device and an air control device; the water supply device mainly includes: a water pipe connected to the water control device outside the sound insulation box, and a water nozzle with an inner diameter of 0.86mm installed at the end of the water pipe and inserted into the interior of the open box; the air supply device mainly includes: an air pipe connected to the air control device, and an air nozzle with an inner diameter of 1.9mm installed at the end of the air pipe and inserted into the interior of the open box;

[0016] In a specific embodiment, the water supply device controls the connection and opening and closing with the water storage bottle through the second solenoid valve, and the air supply device controls the connection and opening and closing with the air pump through the first solenoid valve; the first solenoid valve and the second solenoid valve are connected to the control computer via a relay and an NI data acquisition card and are controlled; the animal motion sensing device is a needle-shaped infrared sensor, which is installed in parallel with the water and air supply device, specifically, inserted into the bright box through a hole outside the soundproof box, located opposite the dark box, and at the end of the bright box. The animal motion sensing device is also connected to the processing control system outside the soundproof box, and is responsible for sensing the occurrence of the experimental animal licking water.

[0017] In the present invention, the animal position sensing device mainly includes: two pairs of matrix-based infrared sensors (ESR-22N with STC-25ML, Schroeder), including a first matrix-based infrared sensor and a second matrix-based infrared sensor; the transmitter and receiver of the first matrix-based infrared sensor are respectively located on the two side boxes in the middle of the dark box, and the transmitter and receiver of the second matrix-based infrared sensor are respectively located on the two side boxes at the junction of the light box and the dark box; the sensor signals of the first matrix-based infrared sensor and the second matrix-based infrared sensor are controlled and processed via a relay outside the soundproof box, an NI data acquisition card, and a MATLAB program in a control computer;

[0018] The innovation of the present invention is that the position status and movement speed of the experimental animal are monitored by combining the first matrix of infrared sensors and the second matrix of infrared sensors; the first matrix of infrared sensors is used to detect whether the experimental animal is completely located in the dark box, and the second matrix of infrared sensors is used to detect whether the experimental animal passes between the light box and the dark box.

[0019] Specifically, the signal fluctuations of the first matrix of infrared sensors and the second matrix of infrared sensors include the following situations:

[0020] 1) If the animal crosses the boundary between the light box and the dark box, the second matrix of infrared sensors will detect a signal fluctuation. If the animal further moves into the dark box (i.e., its body completely enters the dark box), the first matrix of infrared sensors will detect a signal. If the second matrix of infrared sensors senses a signal fluctuation first, followed by a signal fluctuation from the first matrix of infrared sensors, it is determined that the animal has completely entered the dark box, and the direction of movement is from the external light box into the dark box.

[0021] 2) If the first matrix of infrared sensors senses a signal fluctuation first, and then the second matrix of infrared sensors senses a signal fluctuation, it is determined that the animal has completely walked out of the dark box, and the direction of movement is from the dark box to the external light box;

[0022] 3) If the animal is between the dark box and the light box (i.e., it has not completely walked out of or entered the dark box), and only the second matrix infrared sensor senses the signal, while the first matrix sensor does not detect the signal, the trial will not begin;

[0023] By using the signal fluctuations of the first matrix of infrared sensors and the second matrix of infrared sensors, the state of the experimental animals entering and exiting the dark box can be accurately detected, and whether the animals have completely entered the dark box can be accurately determined.

[0024] At the same time, the speed of the signals from the first matrix infrared sensor and the second matrix infrared sensor is calculated, so that the speed of the experimental animals entering and leaving the room can be monitored.

[0025] During the actual training process, experimental animals rarely remain in the position between the light box and the dark box, because the first matrix infrared sensors are set on both sides of the middle of the dark box, and the second matrix infrared sensors are set on both sides of the junction of the light box and the dark box. The actual distance between the first and second matrix infrared sensors is about 5 cm, which is less than the body length of a normal adult experimental animal. At the same time, in order to avoid the occurrence of very rare cases, during the initial adaptation stage of the dark box, it is important to train the animals to completely enter the dark box in each trial to reduce the situation where the animals are located between the two sensors.

[0026] Furthermore, by calculating the time sequence of the first and second matrix infrared sensors generating signals, the movement speed of the experimental animal can be further calculated, thereby realizing fully automatic monitoring of the experimental animal status.

[0027] A speaker (T34B-4Ω, Bliesma) and a lighting lamp are provided on the top of the soundproof box; wherein, the speaker circuit is connected to the power amplifier outside the soundproof box, and the sound playing and shutting are controlled by the processing control system; the lighting lamp is installed on the top of the soundproof box.

[0028] The air control device includes: an air pump (ARP4DC12V, Yuanxun) with an air supply range of 0-2.3 bar, and a first solenoid valve (KVE33PL12FP3Q201, Kamoer); an air duct connects the air pump to the inlet of the first solenoid valve, and an air duct connects the outlet of the first solenoid valve to the air nozzle; air in the air pump is pumped into the first solenoid valve through the air duct, and the first solenoid valve remains closed unless triggered; the first solenoid valve is connected to the relay (RISYM); a MATLAB program in the control computer receives a signal from the animal motion sensing device, determines a behavior error, and controls the relay to open the first solenoid valve, thereby causing the air nozzle to blow once;

[0029] The water control device includes: a water storage bottle and a second solenoid valve (KVE33PL12FP3Q201, Kamoer); the water storage bottle and the inlet of the second solenoid valve are connected via a water conduit, and the outlet of the second solenoid valve is connected to the water faucet via a water conduit; water in the water storage bottle enters the second solenoid valve through the water conduit, and the second solenoid valve is always closed unless triggered; the second solenoid valve is connected to the relay, and when the MATLAB program in the control computer determines that the behavior is correct, the relay controls the second solenoid valve to open, thereby achieving a single water supply from the water faucet;

[0030] The start and stop of the water and gas control device are all comprehensively determined and controlled by the sensor signal-assisted program code, which is the innovation of the present invention and realizes free training and monitoring of experimental animals.

[0031] In the processing and control system, the relay is connected to the NI data acquisition card (NI USB-6001). The NI data acquisition card will collect and convert the signal of the animal motion sensing device and transmit it to the MATLAB program of the control computer. The program will further determine whether the behavior is correct or not, and then transmit the corresponding action signal to the relay through the data acquisition card. The relay controls the solenoid valve to supply water and air.

[0032] The auditory perception training device is equipped with a corresponding MATLAB program. The training program can firstly compile and generate specific sound types for training, further enriching the composition of target sounds, such as setting different frequency combinations, frequency ranges, sound durations, sound loudness, time intervals between different sounds, etc.; at the same time, a sound library consisting of multiple different sound types is established, which allows for greater freedom in sound settings, and the program can randomly play all sounds in the sound library.

[0033] Secondly, the training program combines with relays and NI data acquisition cards to achieve fully automatic detection of the animal's position and movements. During the training process, signals of the animal's position, the correctness of its behavior, and the mechanical action of watering and airing are received and collected by the NI data acquisition card via relays. The corresponding signal data is then received and determined by the MATLAB program, and subsequent watering or air blowing actions are performed.

[0034] When the needle-shaped infrared sensor fixed in parallel with the water spout senses that the animal is licking the water spout, the needle-shaped infrared sensor will transmit the signal to the MATLAB program through the NI data acquisition card. The signal is then judged by the MATLAB program to determine whether the conditions are correct, and the program outputs the subsequent corresponding action signal; that is, when the target sound is played, if the animal walks out of the dark box within the specified time and licks the water spout within the reaction time window, the program automatically determines that the action is correct, counts Hit+1 on the program interface, and controls the second solenoid valve to open and give a water reward; when the target sound is played, if the mouse does not lick the water spout within the reaction time window, the program automatically determines that the action is wrong, controls the first solenoid valve to open and gives a blowing penalty, and counts Miss+1 on the program interface;

[0035] If the animal does not lick the water when the non-target sound is played, the program automatically determines that the behavior is correct and counts "Correct Reject" + 1 on the program interface. If the animal exits the dark box within the specified time and licks the water spout within the reaction time window when the non-target sound is played, the behavior is considered incorrect, and the first solenoid valve is controlled to open to impose a puff penalty, and counts "False Alarm" + 1 on the program interface. The above signal judgments are all automatically received and controlled by the program.

[0036] Finally, the program automatically generates a result output file after each day of training. The output file summarizes: the frequency of sound playback for each trial, the duration of sound playback, the time spent in the dark box, the reaction time window, the time from sound playback to behavior occurrence, and the statistics of the behavior that occurred in each trial. The above content is a substantial innovation compared to existing technologies.

[0037] Based on the above device, the present invention also proposes a non-therapeutic, painless, non-invasive perceptual training method for mice to stimulate auditory cortical plasticity through free training, which includes the following four training stages:

[0038] Phase 1: Pre-experimental phase, mice were deprived of water for 3 days and petted for 10 minutes every day to allow them to adapt to the experimenters;

[0039] Phase 2: Adaptation

[0040] (1) Free adaptation stage: The mice moved freely in the runway. There was no sound stimulation or punishment during this stage, and the mice could lick the water freely. Every time the needle-shaped infrared sensor (animal motion sensing device) sensed a lick, the second solenoid valve controlled the release of a drop of water (4 μl / time). This stage was trained for 1 day and lasted for 20 minutes.

[0041] (2) Initial adaptation stage of dark box:

[0042] The first 4 min of training for each animal was for acclimation training, and the results were not included in the final statistics;

[0043] In this stage, the waiting time (Waiting time) and the response window (Response window) are set to 4 s, that is, the mouse needs to completely enter the dark box and wait for 4 s. Then the speaker will give a target sound (high-frequency pure tone) of 16000 Hz. The mouse must walk out of the dark box within 4 s after the sound is sounded, and lick the water spout through the runway to get a water reward (4 μl / time), which is recorded as a correct behavior (Hit), and the trial ends; then the mouse must learn to return to the dark box and wait before the next trial begins; if the mouse does not complete the licking action, the behavior is recorded as an error (Miss); in this stage, the air puff time (Air puff time) is set to 0 s, that is, there is no air puff penalty.

[0044] Training was conducted twice daily, with 125 trials per session, for a total of 250 trials. The target sound accuracy (H) (%) was calculated as Hit / total target frequency sound production. A target sound accuracy of ≥70% was achieved for two consecutive days before proceeding to the next stage.

[0045] Phase 3: Coupling the two sound phases

[0046] The first 4 min of training for each animal was the adaptation phase, and the training results were not included in the final statistics;

[0047] In this stage, the waiting time and response window were set to 4 s, the same as in the previous stage. However, an air puff penalty was added, with an air puff time of 1 s. The loudspeaker played the target sound (high-frequency sound) of 16000 Hz and the non-target sound (low-frequency sound) of 6964 Hz in a pseudo-random ratio of 5:5 (a difference of 1.2 octaves).

[0048] The octave (octave, oct) refers to the logarithm of the frequency ratio (or pitch ratio) of two sounds with base 2, and the mathematical expression is f2 / f1=2 n , where f1 is the reference frequency, f2 is the signal frequency for which the octave number is to be determined, and n is the octave number, which can be either an integer or a decimal. The above 16000Hz has a frequency difference of 1.2 octaves, which can be calculated by 16000÷2 (1.2) About 6964Hz.

[0049] Based on the type of sound and the different behaviors of the mice, the results were divided into four situations:

[0050] Hit, that is, when the target sound was played, the mouse walked out of the dark box within 4 s and licked the water spout within the 4 s reaction time window;

[0051] Miss, that is, when the target sound was played, the mouse did not lick the water spout within the 4-s reaction time window, and was given a 1-s air-blowing penalty;

[0052] Correct Reject, that is, when the non-target sound was played, the mouse did not lick the water, which meant that the behavior was correct;

[0053] False Alarm: when the non-target sound was played, if the mouse walked out of the dark box within 4 s and licked the water spout within the 4 s reaction time window, it was considered a behavioral error and was punished with a 1 s air puff;

[0054] The mouse then had to return to the dark box and wait before the next trial began;

[0055] Training was conducted twice daily, with 125 trials per session, for a total of 250 trials. The target sound accuracy rate (H(%)) was calculated as Hit / total target frequency sound production, and the non-target sound error rate (F(%)) was calculated as False Alarm / total non-target frequency sound production. The overall performance score (P(Performance,%)) was calculated as HH×F. If the P value was ≥70% and maintained for two consecutive days, the participants moved on to the next stage.

[0056] The fourth stage: training stage, gradually shortening the discrimination gap between target sounds and non-target sounds.

[0057] Three difficulty levels are set in the entire training process, 1.2 oct, 0.8 oct, and 0.6 oct, that is, the interval between the target sound and the non-target sound is gradually shortened, and the training difficulty is gradually progressively from easy to difficult; 1.2 oct has been considered to be a relatively obvious difference between the two sounds in previous fixed training, and is relatively easy to distinguish, and is used in the early stage of training; 0.8 and 0.6 oct are set later, with the purpose of setting difficulty for the training animals while not setting the difficulty too difficult to exceed the animal's limit; preliminary experiments have found that animals can also learn quickly when entering 0.6 oct after learning the 0.8 oct stage. In the specific implementation process, based on the training purpose of improving plasticity and restoring frequency tuning ability to achieve the method of the present invention, it is generally sufficient to set 0.6 oct; in another specific implementation process, if time is sufficient, it is also possible to consider setting a difficulty of 0.4 oct to further improve the training effect;

[0058] (1) 0.8 octave difference: target sound is 13929 Hz, non-target sound is 8000 Hz, waiting time is 4 s, response window is 4 s, air puff time is 1 s, and the sounds are played pseudo-randomly in a 5:5 ratio. The judgment method and number of training times are the same as above.

[0059] (2) 0.6 octave difference: set the target sound to 12996 Hz, the non-target sound to 8574 Hz, the waiting time to 4 s, the response window to 4 s, the air puff time to 1 s, and perform pseudo-random playback at a ratio of 5:5. The judgment method and number of training times are the same as above.

[0060] At the end of each day's training, the amount of water consumed was counted based on the results. The amount of water consumed by each mouse per day did not exceed 1 ml, and the missing amount was accurately supplemented after training.

[0061] The method of the present invention is an auditory perception training method for non-invasive free mouse training to stimulate the plasticity of the auditory cortex. In the past, the common mouse training paradigm was often an invasive fixation method. Usually, the mouse was anesthetized, the scalp was cut open, and a fixation plate was glued to the mouse skull using dental cement and glue, and then fixed with screws. The mouse was fixed in front of the water spout throughout the entire training process, and the behavioral action was determined by whether it stretched out its tongue to lick the water spout. In the present invention, the animal does not need to undergo any traumatic treatment, and it does not need to be fixed when placed in the perceptual training device, and it always maintains a free movement state.

[0062] In the training method of the present invention, during the initial adaptation phase of the dark box, mice are first taught to wait in the dark box for a specified time. A target sound (high-frequency sound) is given as an instruction. Upon hearing the sound, mice must exit the dark box and perform an action within a reaction time window. Finally, they must return to the dark box and wait for the next sound. This completes the establishment of a conditioned reflex. Training is conducted twice a day, with 125 trials each time, for a total of 250 trials. The next phase is initiated after the accuracy rate reaches 70% and is maintained for two days.

[0063] In the training method of the present invention, in the two-sound coupling stage, two high-frequency and low-frequency sounds are given as instructions, so that the animal learns to distinguish the two sounds in this stage. That is, after the target sound (high-frequency sound) sounds, the animal makes a licking action within the reaction time window, and then there will be a water reward, that is, Hit+1; if the target sound sounds but the action is not made within the reaction time window, the animal will receive a 1s air blowing penalty, and Miss+1 is recorded; if the non-target sound sounds and the animal does not make an action, it means that the behavior is correct, and Correct Reject+1 is recorded; if the non-target sound sounds and the animal licks the water spout, it means that the behavior is wrong, and False is recorded Alarm + 1; training was conducted twice daily, 125 trials each time, for a total of 250 trials; after training, each animal's target sound accuracy rate (H(%) = Hit / total number of target frequency sounds) and non-target sound error rate (F(%) = FalseAlarm / total number of non-target frequency sounds were calculated; and each animal's overall performance score (P (Performance,%)) was calculated as HH × F; if the P value for two consecutive days was ≥ 70%, the animal entered the next stage.

[0064] In the training method of the present invention, during the training phase, the gap between the target sound and the non-target sound is gradually shortened within the animal's sound frequency perception range, with the pitch interval being reduced to 13929 Hz for the target sound and 8000 Hz for the non-target sound, a difference of 0.8 octaves, as an instruction. This allows the animal to learn to distinguish two more similar sounds in this phase, thereby increasing the difficulty. Training is conducted twice a day, with 125 trials each time, for a total of 250 trials. After training, each animal's target sound accuracy rate (H(%)) = Hit / total number of target frequency sounds, and non-target sound error rate (F(%) = False Alarm / total number of non-target frequency sounds are calculated. The overall performance score (P(Performance,%)) = HH × F is calculated for each animal. If the P value for two consecutive days is ≥ 70%, the animal enters the next phase.

[0065] In the training method of the present invention, during the training phase, the interval between the target sound and the non-target sound is further shortened, and the pitch interval is reduced to 12996 Hz for the target sound and 8574 Hz for the non-target sound, with a difference of 0.6 octaves. This is used as an instruction to allow the animal to learn to distinguish two more similar sounds in this phase, thereby increasing the difficulty. Training is conducted twice a day, with 125 trials each time, for a total of 250 trials. After training, each animal's target sound accuracy rate H (%) = Hit / total number of target frequency sounds, and non-target sound error rate F (%) = False Alarm / total number of non-target frequency sounds are calculated. The overall performance score P (Performance, %) for each animal is = HH × F. If the P value for two consecutive days is ≥ 70%, the animal is considered to have completed training.

[0066] In the method of the present invention, the animal is placed in a perception training device for training.

[0067] In the method of the present invention, a computer and device hardware are connected through a controller to convert computer language into hardware control signals; sounds are played through a speaker; correct training animals are rewarded by spraying water; animal triggers are received through an infrared sensor; and the trained animals are punished through a blowing device.

[0068] In the method of the present invention, the entire adaptation and training phase is controlled within 20 days. First, the training time cannot be too long, as mice themselves have limited endurance for training time; but the training time cannot be too short either. The most important thing about perceptual training is that it can successfully induce brain plasticity.

[0069] The beneficial effects of the present invention include:

[0070] Compared with previous behavioral devices used for sensory training or decision-making of experimental animals, animals are usually surgically treated in advance, that is, the top of the skull of the mouse is exposed after anesthesia, a stainless steel restraint is fixed with tissue glue, and the steel is completely fixed with dental cement. After the dental cement is completely dry, the mouse is housed in a single cage, and routine care is performed until the mouse recovers to its previous level of movement, and then auditory behavior experiments are performed. In this method, the animal needs to undergo anesthesia and surgery, with an incision on the head. The animal is fixed throughout the training process and can only move by licking with its tongue. In addition, during the training process, since the animal resists after being fixed, the fixed restraint falls off, and the animal needs to be anesthetized again for surgical installation. The present invention improves the previous training method. First, the animal does not need to undergo pre-anesthesia and surgery, and there are no incisions. Second, the animal is completely free to train on the runway, following the nature of the experimental animal. The start and end of training are determined by the experimental animal itself, providing a non-invasive and free cognitive training method.

[0071] Compared with the existing Chinese invention patent CN111700000B, the difference is that: the device in the existing Chinese invention patent is used for auditory training of rats, and the activity area is a large transparent open field; while in the present invention, it is mainly aimed at mice that are smaller than rats, and a straight runway box adapted to the size of mice is set, and the runway length is moderate, which will not make the mice consume too much physical energy during the training process; and the straight runway limits the direction of movement of the mice each time, and has a directional effect on the mice to make movements in the light box after walking out of the dark box, thereby avoiding excessive freedom of the mice and distraction, which affects the training results; the training method in the existing Chinese invention patent is in the process After the program is started, the sound will continue to play at a fixed time interval. However, after the program is started in the present invention, the sound playback is determined by the mouse, that is, the sound will not be played and the trial will not start until the mouse has waited for a specified time in the dark box. The training method in the existing Chinese invention patent uses specially prepared micro-food particles as rewards for correct behavior of rats. Since mice have a small food intake and a slow eating speed, they are not suitable for food particles as a reward method. Therefore, the present invention uses water as a reward, which is more in line with the characteristics of the training objects. The experimental objects in the present invention, namely mice, have much lower intelligence and motor ability than rats. Therefore, in the training method, The training method is simpler and easier to train. The existing Chinese invention patent uses a light as a reminder for rats to behave incorrectly. However, in the present invention, since the animals are free to move during the training process and the light is fixed, there are many cases where the animals cannot receive the light reminder due to their activities after each behavior is wrong, and the punishment is not strong enough. Therefore, the present invention uses an air blowing punishment, and the air nozzle is located above the water nozzle. After the mice behave incorrectly, they can be directly punished by the air blowing. The program in the present invention has greater freedom. The program uses a sound library as a unit to encode and generate multiple sound files for random playback, which can further enrich the composition of a single sound file. For example, For example, different frequency combinations, frequency ranges, sound durations, sound loudness, time intervals between different sounds, etc. can be set to enrich the training content. The adult brain still has a considerable degree of plasticity, although it is relatively limited, but it can still be used for sensory input through cognitive training, and through the interaction with behavior, the activity level and pattern of the cortical circuit can be further changed. That is, the auditory cognitive training described in the present invention can effectively improve the brain plasticity of adult animals, and by changing the training sound, it can further improve the frequency selectivity, repetition rate following ability, and loudness discrimination ability of the animal's auditory cortex neurons, and has a wide range of application prospects. It is also universally applicable in many neurological problems and helps to improve the functional abnormalities of the animal's auditory cortex. For example, in autistic animals, the frequency selectivity of auditory cortical neurons is reduced, and the ability to process sound information is impaired. Cognitive training can improve the functional abnormalities of the auditory cortex caused by animal autism. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without paying any creative work.

[0073] Figure 1 Diagram of the apparatus for the free-listening cognitive training paradigm in mice.

[0074] Figure 2 Schematic diagram of behavioral assessment in the auditory perception training paradigm for free mice.

[0075] Figure 3 It is a structural schematic diagram of the auditory perception training device of the present invention.

[0076] Figure 4 Schematic diagram of the auditory perception training paradigm for free mice that can improve the frequency receptive field in the auditory cortex of autistic mice.

[0077] Figure 5 Figure 2: Experimental results showing that the frequency selectivity of auditory cortical neurons in autistic mice can be widely improved through an auditory perception training paradigm in free mice.

[0078] Figure 6 Comparison of auditory perception training performance in the valproic acid-treated mice. DETAILED DESCRIPTION

[0079] The invention is further described in detail with reference to the following specific examples and accompanying drawings. The processes, conditions, experimental methods, etc. for implementing the present invention, except for those specifically mentioned below, are common knowledge and common common sense in the art and are not particularly limited by the present invention.

[0080] The present invention provides a non-therapeutic, non-invasive, and painless auditory perceptual training device and method for stimulating auditory cortical plasticity through free training. The present invention also provides the use of the training device for inducing auditory cortical plasticity and perceptual training. Unlike commonly used training paradigms, the present invention establishes a non-invasive, painless, free training paradigm. This approach aims to enhance the frequency selectivity of auditory cortical neurons through sound frequency discrimination training, and has broad application prospects.

[0081] The present invention also provides a novel, non-invasive, free method for auditory perception training of mice for non-therapeutic purposes, comprising the following four training stages:

[0082] Phase 1: Pre-experimental phase, mice were deprived of water for 3 days and petted for 10 minutes every day to allow them to adapt to the experimenters;

[0083] Phase 2: Adaptation

[0084] (1) Free adaptation stage: The mice moved freely in the runway. There was no sound stimulation or punishment in this stage, and the mice could lick water freely. Every time the needle-shaped infrared sensor sensed a lick, the second solenoid valve controlled the release of a drop of water (4 μl / time). This stage was trained for 1 day and lasted for 20 minutes.

[0085] (2) Initial adaptation stage of dark box:

[0086] The first 4 min of training for each animal was for acclimation training, and the results were not included in the final statistics;

[0087] In this stage, the waiting time (Waiting time) and the response window (Response window) are set to 4 s, that is, the mouse needs to completely enter the dark box and wait for 4 s. Then the speaker will give a target sound (high-frequency pure tone) of 16000 Hz. The mouse must walk out of the dark box within 4 s after the sound is sounded, and lick the water spout through the runway to get a water reward (4 μl / time), which is recorded as a correct behavior (Hit), and the trial ends; then the mouse must learn to return to the dark box and wait before the next trial begins; if the mouse does not complete the licking action, the behavior is recorded as an error (Miss); in this stage, the air puff time (Air puff time) is set to 0 s, that is, there is no air puff penalty.

[0088] Training was conducted twice daily, with 125 trials per session, for a total of 250 trials. The target sound accuracy (H) (%) was calculated as Hit / total target frequency sound production. A target sound accuracy of ≥70% was achieved for two consecutive days before proceeding to the next stage.

[0089] Phase 3: Coupling the two sound phases

[0090] The first 4 min of training for each animal was the adaptation phase, and the training results were not included in the final statistics;

[0091] In this stage, the waiting time and response window were set to 4 s, the same as in the previous stage. However, an air puff penalty was added, with an air puff time of 1 s. The loudspeaker played the target sound (high-frequency sound) of 16000 Hz and the non-target sound (low-frequency sound) of 6964 Hz in a pseudo-random ratio of 5:5 (a difference of 1.2 octaves).

[0092] Based on the type of sound and the different behaviors of the mice, the results were divided into four situations:

[0093] Hit, that is, when the target sound was played, the mouse walked out of the dark box within 4 s and licked the water spout within the 4 s reaction time window;

[0094] Miss, that is, when the target sound was played, the mouse did not lick the water spout within the 4-s reaction time window, and was given a 1-s air-blowing penalty;

[0095] Correct Reject, that is, when the non-target sound was played, the mouse did not lick the water, which meant that the behavior was correct;

[0096] False Alarm: when the non-target sound was played, if the mouse walked out of the dark box within 4 s and licked the water spout within the 4 s reaction time window, it was considered a behavioral error and was punished with a 1 s air puff;

[0097] The mouse then had to return to the dark box and wait before the next trial began;

[0098] Training was conducted twice daily, with 125 trials per session, for a total of 250 trials. The target sound accuracy rate (H(%)) was calculated as Hit / total target frequency sound production, and the non-target sound error rate (F(%)) was calculated as False Alarm / total non-target frequency sound production. The overall performance score (P(Performance,%)) was calculated as HH×F. If the P value was ≥70% and maintained for two consecutive days, the participants moved on to the next stage.

[0099] The fourth stage: training stage, gradually shortening the discrimination gap between target sounds and non-target sounds.

[0100] (1) 0.8 octave difference: target sound is 13929 Hz, non-target sound is 8000 Hz, waiting time is 4 s, response window is 4 s, air puff time is 1 s, and the sounds are played pseudo-randomly in a 5:5 ratio. The judgment method and number of training times are the same as above.

[0101] (2) 0.6 octave difference: set the target sound to 12996 Hz, the non-target sound to 8574 Hz, the waiting time to 4 s, the response window to 4 s, the air puff time to 1 s, and perform pseudo-random playback at a ratio of 5:5. The judgment method and number of training times are the same as above.

[0102] At the end of each day's training, the water intake of each mouse was counted based on the results. The water intake of each mouse did not exceed 1 ml per day, and the missing amount was accurately supplemented after training.

[0103] The present invention provides an auditory perception training device for non-invasive free mouse training to stimulate auditory cortex plasticity for non-therapeutic purposes, the device comprising: a soundproof box and a training device body therein, and a processing control system located outside the soundproof box;

[0104] The training device body inside the soundproof box includes: a training box body, a water and air supply device, an animal motion sensing device, an animal position sensing device, a speaker, and a lighting lamp; the training box body includes a light box and a dark box, and the animal position sensing device is installed at the relative position of the side wall at the junction of the light box and the dark box, and at the relative position of the side wall of the dark box;

[0105] The processing and control system located outside the soundproof box includes: a water and gas control device, a power amplifier and a control computer.

[0106] The light box is a white, opaque box with an open top; the dark box is a black, opaque box with a closed top; the light box and the dark box are interconnected and unobstructed, and the dark box is provided with a movable back door for the entry and exit of experimental animals;

[0107] The water and air supply device and the animal motion sensing device are inserted into the light box through holes outside the soundproof box. The insertion position is located opposite to the dark box and at the sealing plate at the end of the light box.

[0108] The water and air supply device includes a water supply device and an air supply device, and the water and air control device includes a water control device and an air control device; the water supply device includes a water pipe connected to the water control device outside the sound insulation box, and a water nozzle installed at the end of the water pipe and inserted into the interior of the open box; the air supply device includes an air pipe connected to the air control device, and an air nozzle installed at the end of the air pipe and inserted into the interior of the open box;

[0109] and / or,

[0110] The animal motion sensing device is inserted into the bright box through a hole outside the soundproof box, and is connected to the processing control system outside the soundproof box, and is responsible for sensing the occurrence of the experimental animal's licking action.

[0111] The air control device includes: an air pump and a first solenoid valve; the first solenoid valve is controlled by the control computer to control the connection and opening and closing of the air supply device and the air pump;

[0112] The water control device includes a water storage bottle and a second electromagnetic valve; the second electromagnetic valve is controlled by the control computer to control the connection and opening and closing of the water supply device and the water storage bottle.

[0113] The animal position sensing device includes: two pairs of matrix-based infrared sensors; wherein the transmitter and receiver of the first matrix-based infrared sensor are respectively located on two side boxes in the middle of the dark box, and the transmitter and receiver of the second matrix-based infrared sensor are respectively located on two side boxes at the junction of the light box and the dark box; the first matrix-based infrared sensor and the second matrix-based infrared sensor are controlled by a control computer outside the soundproof box;

[0114] The first matrix of through-beam infrared sensors is used to detect whether the experimental animal is completely located in the dark box, and the second matrix of through-beam infrared sensors is used to detect whether the experimental animal passes between the light box and the dark box.

[0115] The speaker and the lighting are installed on the top of the soundproof box; wherein, the circuit of the speaker is connected to the power amplifier outside the soundproof box, and the sound playback and shutoff are controlled by the processing control system; the lighting is used for lighting in the soundproof box.

[0116] Specifically, if Figure 1 As shown, A, the training device is located in a soundproof shielded box. The main runway is made of opaque plexiglass and consists of a milky white open box (open top) and a dark box (with a cover). B, the design dimensions of the main runway include a 25cm long, 8cm wide, and 25cm high open box section and a 10cm long, 8cm wide, and 25cm high dark box section. The water nozzle and the needle-shaped infrared sensor are intertwined and fixed, inserted through a hole punched outside the box at the end of the runway, on the left side. The air nozzle is located below the water nozzle and also inserted through a hole punched outside the box. Matrix infrared sensor B is inserted through a hole punched at the junction of the open and dark boxes, and matrix infrared sensor A is inserted through a hole punched in the middle of the dark box. A movable rear door is installed on the right side of the runway (inside the dark box) to facilitate training operations and animal catching.

[0117] The auditory perception training device and method of the present invention can demonstrate the following:

[0118] 1. Perceptual training affects the auditory discrimination ability of mice.

[0119] In the early stage of perceptual training, mice would not wait for the specified time in the dark box, nor would they walk out of the dark box according to the sound played as a command, and then perform behavioral actions; in the later stage of training, mice learned to wait for the specified time and respond to the target sound, but not to the non-target sound ( Figure 2 ).

[0120] Specifically, Figure 2 Schematic diagram of behavioral judgment in the auditory perception training paradigm for free mice; in the two coupled sound stages, after the target sound (high-frequency sound) sounds, the animal will only receive a water reward if it licks the water within the reaction time window, i.e., Hit; if the target sound sounds but the animal does not take action within the reaction time window, it will receive a 1-s air-blowing penalty and be recorded as Miss; if the animal does not take action when the non-target sound sounds, it means the behavior is correct, recorded as Correct Reject; if the animal licks the water spout when the non-target sound sounds, it means the behavior is incorrect, recorded as False Alarm.

[0121] 2. Perceptual training can widely improve the function of the auditory cortex.

[0122] It has been shown that changes in the frequency receptive field of auditory cortical neurons can affect an individual's auditory discrimination behavior. The frequency receptive field of auditory cortical neurons primarily refers to the range of frequency and intensity of sound that auditory cortical neurons perceive, as determined by hardware systems that alter the frequency and intensity of sound stimuli. This description of the frequency receptive field of auditory cortical neurons can provide insights into their function. Figure 4 Schematic diagram of the auditory perception training paradigm for free mice that can improve the frequency receptive field of the auditory cortex of autistic mice. Figure 4 Figure A shows a frequency tuning curve diagram, which labels indicators such as characteristic frequency (CF), minimum threshold (MT), and suprathreshold bandwidth (Bandwidth, BW). Figure 4 Figure B shows examples of the frequency tuning characteristics of auditory cortical neurons in the control, valproate-treated, and valproate-trained groups, respectively. The frequency receptive fields of the valproate-treated mice were wider than those in the control group, while those of the valproate-trained mice were narrower.

[0123] By outlining the frequency receptive field, we can get the frequency tuning curve of the auditory cortex neurons. Generally, the horizontal axis (frequency) corresponding to the lowest point of the frequency tuning curve is used as the characteristic frequency of the neuron, and the width of the curve at 10, 20, and 30 dB above the lowest point (BW10, 20, 30) is used as an indicator to describe the size of the receptive field. Figure 5 The figure shows the experimental results that the auditory perception training paradigm of free mice can widely improve the frequency selectivity of auditory cortical neurons in autistic mice. Statistical results of the bandwidth (BW10, 20, 30) and threshold of the frequency receptive field of all recorded neurons at 10, 20, and 30 dB above the threshold. Data are expressed as Mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; Specifically, by using the recorded neurons, it was found that the BW10, 20, and 30 of the receptive field of the auditory cortical neurons of autistic mice treated with valproic acid were higher than those of The BW10, 20, and 30 receptive fields of auditory cortical neurons in the valproate-treated mice after perceptual training returned to normal levels (P(control group vs. valproate-treated group) <0.001, P(control group vs. valproate-trained group) = 0.0363, P(valproate-treated group vs. valproate-trained group) <0.001, one-way ANOVA). This suggests that this training paradigm can broadly improve individual auditory cortical function.

[0124] 3. Perceptual training can improve the abnormal function of the auditory cortex in autistic animals.

[0125] Most studies have shown that Autism Spectrum Disorder (ASD), as a neurodevelopmental disorder, usually presents with social disorders, language communication defects, and repetitive stereotyped behaviors. Perceptual abnormalities are widely present in 90% of autistic patients, among which auditory system abnormalities are the most common. The auditory system plays an important role in the development of children's language, and language dysfunction will affect social skills to a certain extent. Therefore, among the sensory abnormalities in autism, the study of the function of the auditory system is particularly important.

[0126] Growing evidence suggests that exposure to anti-epileptic drugs during pregnancy increases the risk of autism. Among them, valproic acid (VPA) is a drug widely used to treat epilepsy, migraines, and bipolar disorder. Exposure during specific gestational periods can cause fetal developmental defects, including skeletal malformations, behavioral disorders, and cognitive deficits. Autism mouse models established through prenatal injection of valproic acid exhibit stable autism-like features of impaired social interaction, abnormal sensory responses, and anxiety-like and repetitive behaviors. It also affects the development of the auditory cortex, leading to abnormal auditory function in adulthood.

[0127] For this part of the experiment, a valproate mouse model was first established. Eight-week-old male and female mice were purchased and cage-bred in a 1:2 ratio. 1g of VPA (Sigma) was weighed and dissolved in saline to prepare a 250mg / ml VPA saline solution. On day 12.5 of gestation (when a vaginal plug was observed, it was gestational day 0.5), the female mice were intraperitoneally injected with 600mg / kg body weight of VPA saline solution. All male offspring born from the pregnant mice were designated as the valproate-treated group. The day of birth was designated P0. After 21 days of birth, the mice were separated and caged together with a control group (normal C56BL / 6J male mice) until 8 weeks of adulthood, completing this part of the experiment. Mice in the valproate-treated group, trained in this paradigm after P56, were designated the valproate-trained group.

[0128] like Figure 5As shown in the figure, by comparing the frequency receptive field bandwidths of the control group, the valproic acid treated group, and the valproic acid trained group, it can be seen that the bandwidths at 10, 20, and 30 dB above the threshold of the valproic acid group are significantly higher than those of the control group. This suggests that the auditory cortex function of the autistic mice treated with valproic acid is indeed affected, and the frequency selectivity deteriorates. After training with the paradigm of the present invention, by comparing the auditory cortex bandwidth indicators of the valproic acid treated group and the valproic acid trained group, the bandwidths at 10, 20, and 30 dB above the threshold of the valproic acid trained group are significantly lower than those of the valproic acid treated group. This suggests that training can significantly improve the width of the frequency receptive field of auditory cortical neurons in valproic acid autistic mice, increase the frequency selectivity of auditory cortical neurons, and improve auditory cortical function.

[0129] 4. Animals' learning of sound frequency discrimination during free training

[0130] like Figure 6 Figure 2 shows the training progress of four animals in the valproic acid-treated group at different stages of training. The target sound accuracy rate (H)(%) is calculated as Hit / total number of target frequency sounds, and the non-target sound error rate (F)(%) is calculated as FalseAlarm / total number of non-target frequency sounds. The overall performance score (P(Performance,%)) is calculated as HH×F. The gray line in the figure represents the animal's performance score for the target sound. The uncolored background represents the two-sound coupling phase, which lasted approximately 16 days and exhibited individual variability. Performance scores increased during the two-sound coupling phase. When performance scores exceeded 70%, the animals entered the next phase, with a colored background, at a training difficulty of 0.8 octs. While individual performance scores may decline due to the increased difficulty of training, they will rise again to over 70% with subsequent training.

[0131] The auditory perception training device and method of the present invention do not require painful or negative stimulation, and the animals will not have negative emotions.

[0132] References

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[0144]

[12] Zhou

[0145] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the appended claims.

Claims

1. An auditory perception training device for non-invasive free-range mouse training to stimulate auditory cortical plasticity for non-therapeutic purposes, characterized in that: The device comprises: a soundproof box and a training device body inside the box, and a processing control system located outside the soundproof box; The training device body inside the soundproof box includes: a training box body, a water and air supply device, an animal motion sensing device, an animal position sensing device, a speaker, and a lighting lamp; the training box body includes a light box and a dark box, and the animal position sensing device is installed at the relative position of the side wall at the junction of the light box and the dark box, and at the relative position of the side wall of the dark box; The processing and control system located outside the soundproof box includes: a water and gas control device, a power amplifier and a control computer.

2. The auditory perception training device according to claim 1, wherein The light box is a white, opaque box with an open top; the dark box is a black, opaque box with a closed top; the light box and the dark box are interconnected and unobstructed, and the dark box is provided with a movable rear door for the entry and exit of experimental animals; The water and air supply device and the animal motion sensing device are inserted into the bright box through holes outside the soundproof box, and the insertion position is located opposite to the dark box and at the sealing plate at the end of the bright box.

3. The auditory perception training device according to claim 2, wherein The water and air supply device includes a water supply device and an air supply device, and the water and air control device includes a water control device and an air control device; the water supply device includes a water pipe connected to the water control device outside the sound insulation box, and a water nozzle installed at the end of the water pipe and inserted into the interior of the open box; the air supply device includes an air pipe connected to the air control device, and an air nozzle installed at the end of the air pipe and inserted into the interior of the open box; and / or, The animal motion sensing device is inserted into the bright box through a hole outside the soundproof box, and is connected to the processing control system outside the soundproof box, and is responsible for sensing the occurrence of the experimental animal's licking action.

4. The auditory perception training device according to claim 3, wherein The air control device includes: an air pump and a first solenoid valve; the first solenoid valve is controlled by the control computer to control the connection and opening and closing of the air supply device and the air pump; The water control device includes a water storage bottle and a second electromagnetic valve; the second electromagnetic valve is controlled by the control computer to control the connection and opening and closing of the water supply device and the water storage bottle.

5. The auditory perception training device according to claim 1, wherein The animal position sensing device includes: two pairs of matrix-based infrared sensors; wherein the transmitter and receiver of the first matrix-based infrared sensor are respectively located on two side boxes in the middle of the dark box, and the transmitter and receiver of the second matrix-based infrared sensor are respectively located on two side boxes at the junction of the light box and the dark box; the first matrix-based infrared sensor and the second matrix-based infrared sensor are controlled by a control computer outside the soundproof box; The first matrix of through-beam infrared sensors is used to detect whether the experimental animal is completely located in the dark box, and the second matrix of through-beam infrared sensors is used to detect whether the experimental animal passes between the light box and the dark box.

6. The auditory perception training device according to claim 1, wherein The speaker and the lighting are installed on the top of the soundproof box; wherein, the circuit of the speaker is connected to the power amplifier outside the soundproof box, and the sound playback and shutoff are controlled by the processing control system; the lighting is used for lighting in the soundproof box.

7. A non-invasive auditory perception training method for stimulating auditory cortical plasticity in free-range mice for non-therapeutic purposes, characterized in that: The method comprises the following stages: Phase I, pre-experimental phase: experimental mice were deprived of water for 3 days and comforted to adapt to the environment; The second stage, the adaptation stage: includes the free adaptation stage and the dark box initial adaptation stage; 1) Free adaptation stage: The experimental mice moved freely without stimulation; after each lick, the second solenoid valve controlled the release of one drop of water. The training lasted one day, a total of 20 minutes. 2) Dark box initial adaptation phase: The experimental mouse enters the dark box and waits for a waiting time before the target sound is sounded. It must exit the dark box and lick water within the reaction time window to receive a reward. "Hit" is set as a correct behavior and "Miss" is set as a behavioral error. Training is performed 250 times per day, and the target sound accuracy rate H is calculated. If H ≥ 70% and is sustained for 2 days, it enters the next phase. The third stage, the coupled two-sound stage, randomly plays the target and non-target sounds and evaluates the behavioral responses of the experimental mice. 250 training sessions are conducted daily, and the accuracy rate (H) of the target sound and the error rate (F%) of the non-target sound are calculated, as well as the overall performance score (P = HH × F). If P ≥ 70% and persists for 2 days, the mice will proceed to the next stage. The fourth stage, the training stage, shortened the resolution gap between the target and non-target sounds by setting the octaves between the target and non-target sounds to 0.8 and 0.6, respectively. The behavioral responses of the experimental mice were evaluated according to the third stage. If P ≥ 70% for two consecutive days, the final training of the experimental mice was considered complete.

8. The auditory perception training method according to claim 7, wherein: The waiting time in the free adaptation phase and the dark box initial adaptation phase is 4 seconds, and the reaction time window is 4 seconds; a 4-minute adaptation phase is also set before training; and / or, In the second stage, the target sound accuracy rate H=Hit / total target frequency sounding times; and / or, In the third stage, the behavioral responses of the mice evaluated include Hit, Miss, Correct Reject, and False Alarm; Hit means that when the target sound is played, the mouse walks out of the dark box within 4 seconds and licks the water spout within the 4-second reaction time window; Miss means that when the target sound is played, the mouse does not lick the water spout within the 4-second reaction time window, and a 1-second air-blowing penalty is given; Correct Reject means that when the non-target sound is played, the mouse does not lick the water, which means the behavior is correct; False Alarm means that when the non-target sound is played, the mouse walks out of the dark box within 4 seconds and licks the water spout within the 4-second reaction time window, which means the behavior is wrong, and a 1-second air-blowing penalty is given; each evaluation result is counted by one; and / or, In the third stage, the target sound accuracy rate H = Hit / total number of target frequency sounds, and the non-target sound error rate F = FalseAlarm / total number of non-target frequency sounds.

9. The auditory perception training method according to claim 7, wherein: The method comprises placing an animal in the auditory perception training device according to any one of claims 1 to 6 for training.

10. The auditory perception training method according to claim 9, wherein: The first solenoid valve for controlling air blowing, the second solenoid valve for controlling water discharge from the faucet, the relay for controlling the start and stop of the solenoid valves, and the NI data acquisition card for providing signals to control the operation of the solenoid valves and performing data acquisition and conversion are connected through an external line controller. Connect the speakers to the amplifier to play the sound; By connecting the water storage bottle and the second solenoid valve and sensing it through the animal motion sensing device, the experimental mice are rewarded for correct actions through the water nozzle; The position of the experimental mice and the direction of their movement in and out of the dark box are detected by a matrix infrared sensor; Give the experimental mice an error warning by connecting the air pump and the air nozzle of the first solenoid valve.

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