USVs-based juvenile mouse model perception quantitative evaluation method and application

Abstract

The invention provides a USVs-based juvenile mouse model sensory perception quantitative evaluation method and application, the evaluation method can evaluate the sensory perception ability of a juvenile mouse model, a juvenile mouse can give out ultrasonic waves in the maternal and infant separation period, cold stimulation lower than a comfortable temperature is performed on the juvenile mouse in the maternal and infant separation period, and the sensory perception ability of the juvenile mouse model can be evaluated. Different sound production expressions caused by cold stimulation are analyzed, and the perceptual ability is evaluated. Based on the evaluation method, firstly, the perceptual ability of the juvenile autism mouse model can be effectively evaluated in the early stage of the disease; secondly, abnormal perception of a neurodevelopment disorder patient taking autism as an example on pain can be effectively simulated; the experiment device is easy to obtain, and the damage to the mouse is small.

Description

Technical Field

[0001] The present invention belongs to the technical field of neurobiological experiments, and particularly relates to a method and application for quantitatively evaluating the perception of an infant mouse model based on USVs. Background Art

[0002] Perception, as the core physiological mechanism for the body to integrate external objective stimuli, is not only the cornerstone of cognitive activities but also an important guarantee for humans to adapt to the environment. In the field of neurodevelopmental disorders, perceptual abnormalities have become a key research direction in clinical research, and their complex manifestations and profound impacts far exceed traditional cognition. Taking autism spectrum disorder (ASD) as an example, the DSM-5 diagnostic criteria innovatively include perceptual abnormalities in the core symptoms and systematically describe the extreme response patterns of patients to sound, light, and touch: some children have a severe vomiting reflex to the sound wave of a hair dryer, while others have no pain response to skin damage. This heterogeneous processing mechanism of the nervous system also shows differences in other neurodevelopmental disorders such as ADHD and intellectual disabilities. Neuroimaging studies have revealed that the functional connection between the prefrontal cortex and the thalamus in children with ASD is abnormal, resulting in the failure of the sensory signal filtering mechanism. This neurobiological basis explains why more than most children with ASD have at least two sensory modality abnormalities, specifically manifested as compulsive tearing of clothing labels or panic reactions to specific frequency sound waves. The behavioral compensation mechanisms caused by perceptual abnormalities often lead to serious consequences. Longitudinal studies have shown that the risk of self-harm behavior in ASD children with tactile hyposensitivity increases several times, typically manifested as repeated hitting hard objects resulting in retinal detachment or persistent finger biting leading to finger bone deformation. These abnormal behaviors are not only manifestations of sensory seeking but may also stem from the stress response of the limbic system to abnormal sensory input.

[0003] The construction of an early intervention system needs to follow the law of the neuroplasticity window period. The American Academy of Pediatrics recommends including sensory processing assessment in the 18-month screening program and using standardized tools such as Sensory Profile 2 for quantitative analysis. Evidence-based medicine has confirmed that early intervention based on the concept of sensory integration (such as the DIR / Floortime model) can improve the sensory abnormalities of ASD children and promote the development of social skills at the same time. The multi-modal intervention programs emphasized in children's rehabilitation-related plans, integrating new technologies such as transcranial magnetic stimulation and virtual reality sensory training, are gradually forming a three-level prevention system.

[0004] Future research urgently needs to break through the interaction mechanism of perception-emotion-cognition and analyze the molecular markers of abnormal sensory processing through multi-omics technologies. The application of new brain imaging technologies such as functional near-infrared spectroscopy (fNIRS) provides the possibility for real-time monitoring of the sensory processing process. This deep integration of basic research and clinical translation will push the rehabilitation of neurodevelopmental disorders into a new era of precision medicine.

[0005] Due to the high homology of its genome with that of humans and its relatively short reproductive cycle, mice are commonly used in the research of brain function, diseases, and mental disorders. In scientific research, human behavior is usually analogized through the behavior of mice. For example, social interaction among mice is used to simulate human social behavior (such as group cooperation, social decision-making in a stressful environment, etc.) to mimic human social disorder characteristics; anhedonia in the depression model can be quantified through the sucrose preference experiment; and the open field test and tail suspension test are used to evaluate anxiety and depression-like behaviors. However, there are significant limitations in the existing behavioral paradigms. Most studies focus on adult mice, lacking detection methods for key neurodevelopmental windows such as infancy or adolescence, and lacking effective detection methods for early-onset neurodevelopmental disorders, which is not conducive to the early diagnosis and treatment of diseases. Summary of the Invention

[0006] To solve the problems existing in the prior art, the present invention provides a method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs and its application, and constructs a method for evaluating the sensory perception ability of a neonatal mouse model. By the characteristic that neonatal mice emit ultrasonic vocalizations (USVs) during mother-infant separation, cold stimulation is simultaneously applied to them during mother-infant separation, and their sensory perception ability is evaluated by analyzing the different USVs caused by cold stimulation.

[0007] To achieve the above object, the technical solution adopted by the present invention is: A method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs, comprising the following steps: Set the temperature to the comfortable breeding environment temperature of the mice. Take each neonatal mouse out of the side of the mother mouse and place it in the test device, record its USVs and behavioral video for a set duration. After the recording is completed, take it out of the test device and put it back beside the mother mouse, and then take out the next neonatal mouse until all neonatal mice are recorded. After sufficient rest beside the mother mouse, then sequentially place the cubs in the test device at a temperature lower than the comfortable temperature, record the USVs and behaviors during cold stimulation for a set duration, and after the end, let the cubs return to room temperature and then put them back beside the mother mouse. Analyze the USVs characteristics of the cubs when receiving cold stimulation at a comfortable temperature and a temperature lower than the comfortable temperature, including: the number of USVs, the average USVs duration, the average USVs main frequency, the average USVs frequency difference, and the average USVs loudness index; compare Shank3b + / + and Shank3b - / - the changes in the USVs characteristic indexes of the mouse cubs at a comfortable temperature and a temperature lower than the comfortable temperature, and obtain the behavioral phenotypes of neonatal autistic mice according to the changes. Use Smart analysis software to analyze the recorded ethological movement videos and compare the differences in ethological indicators of the pups at room temperature and under cold stimulation; Analyze the ethological indicators of mice and the characteristics of USVs to obtain the correlation between the behavioral changes induced by temperature stimulation in neonatal mice.

[0008] Furthermore, each pup was at least 40 min beside the mother during the two tests recording.

[0009] Furthermore, the test device includes a soundproof box, in which a refrigeration plate, an ultrasonic probe and an infrared night vision camera are arranged; the ultrasonic probe and the infrared night vision camera are connected to a computer, and the refrigeration plate is a refrigeration plate with a temperature controller.

[0010] Furthermore, the comfortable temperature is 25 °C, and the cold stimulation temperature lower than the comfortable temperature is 19 °C.

[0011] Furthermore, the set duration is 5 minutes.

[0012] Furthermore, before the test, the cage containing the pups and their mother was placed in an environment with a comfortable temperature, shielded from light and quiet for at least 1 hour for adaptation.

[0013] Furthermore, the cold stimulation lower than the comfortable temperature is in Shank3b The cold stimulation significantly increased the USVs in both the autistic mouse model and the wild-type neonatal mice, suggesting that both types of mice can perceive cold stimulation and express it through USVs; The locomotion of wild-type neonatal mice significantly decreased under cold stimulation lower than the comfortable temperature, while the total locomotion time of autistic mice remained unchanged, indicating their abnormal temperature perception ability; Compared with the comfortable temperature, after cold stimulation lower than the comfortable temperature, the duration of USVs in wild-type neonatal mice was significantly prolonged, the main frequency decreased, the frequency difference increased significantly, and the loudness increased significantly; Shank3b - / - The change of USVs signal in autistic mice after cold stimulation lower than the comfortable temperature was inconsistent with that of wild-type mice, and showed insensitivity to cold stimulation, suggesting that there are obstacles in their perception and expression of perception.

[0014] Furthermore, the ethological indicators include the total locomotion time, total locomotion distance, number of falls and fall time.

[0015] Meanwhile, the application of the method for quantitatively evaluating the sensory perception of the juvenile mouse model based on USVs is provided. Cold stimulation is applied to mouse pups to observe their sensory perception ability. The response of mouse pups to cold stimulation below the comfortable temperature is quantified using USVs, which is used to evaluate the sensory perception ability of neurodevelopmental disorder models, including autism, in the early stage of the disease. This method can not only evaluate the sensory perception ability of the autism mouse model in the early stage of the disease, but also effectively simulate the abnormal perception of temperature by autistic patients. This non-invasive, sensitive, and quantifiable experimental index provides a valuable tool for studying social behavior, emotional expression, and related nervous system disease models.

[0016] The application of the method for quantitatively evaluating the sensory perception of the juvenile mouse model based on USVs can, by comparing the sensory perception abilities of mouse pups of different disease models and combining the research on molecular mechanisms and circuit mechanisms, more deeply understand the pathological basis of neurodevelopmental disorders and assist in finding intervention targets and treatment methods for the diseases. The application of this method can not only provide a scientific basis for the early diagnosis and treatment of neurodevelopmental disorders such as autism, but also promote the research and development of related drugs and intervention measures. By comprehensively analyzing USVs data and molecular biology indicators, the treatment effect can be more accurately evaluated and the treatment plan can be optimized.

[0017] Compared with the prior art, the present invention has at least the following beneficial effects: It can effectively evaluate the sensory perception ability of the juvenile autism mouse model in the early stage of the disease, which is crucial for early diagnosis and intervention. Secondly, this method can effectively simulate the abnormal perception of temperature by neurodevelopmental disorder patients, such as autism, providing a new perspective for the research on the neural mechanisms related to autism symptoms. In addition, the present invention is easy to operate, the experimental device is easy to obtain, and the damage to mice is small, meeting animal ethics. These characteristics make the present invention have a broad application prospect in neuroscience research and disease model evaluation. Description of the Drawings

[0018] Figure 1 For Shank3b + / + and Shank3b - / - Characteristics of cold stimulation USVs and behavioral changes in mouse pups. Detailed Embodiments

[0019] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0020] The present invention provides a method for quantitatively evaluating the sensory perception of a juvenile mouse model based on USVs, comprising the following steps: Set the temperature to the comfortable breeding environment temperature of the mice. Take each young mouse out from beside its mother and place it in the test device, record its USVs and behavioral video for a set duration. After the recording is completed, take it out from the test device and put it back beside its mother, and then take out the next young mouse until all young mice are recorded; After fully resting beside the mother, sequentially place the cubs in the test device at a temperature lower than the comfortable temperature, record the USVs and behaviors during cold stimulation for a set duration, and after the end, let the cubs return to room temperature and then put them back beside their mother; Analyze the ultrasonic vocalization characteristics of the cubs when receiving comfortable temperature and cold stimulation lower than the comfortable temperature, including: the number of USVs, average USVs duration, average USVs main frequency, average USVs frequency difference, average USVs loudness index; compare Shank3b + / + and Shank3b - / - the changes in the basic characteristic indexes of mouse USVs of the mouse cubs at comfortable temperature and lower than comfortable temperature, and obtain the behavioral phenotypes of autistic mice in infancy according to the changes; Use Smart analysis software to analyze the recorded behavioral movement videos, compare the total movement time, total movement distance, number of falls, and fall time of the cubs at room temperature and during cold stimulation, and quantify the differences in the behavioral responses of the cubs to different sensations; Analyze the mouse behavioral indexes and USVs indexes to obtain the correlation between the behavioral changes caused by temperature stimulation in the cub mice. Mice and humans have a high degree of genomic homology, especially in conserved neurodevelopment-related genes such as BDNF and the Shank3 family, which have a high degree of functional conservation, enabling gene editing technology to accurately construct neurodevelopment disorder models; the 21-day gestation period and the reproductive capacity of 6-12 offspring per litter support the design of high-throughput drug screening experiments.

[0021] Example 1 Use the autistic mouse cubs at postnatal day 7 (P7), Shank3b - / - and the littermate control group Shank3b + / + mouse cubs (here is a typical application, and other different autistic mouse models or mouse models with sensory disorders can be used according to the experimental purpose). Before the behavior starts, place the mouse cage containing the young mouse and its mother in a behavioral laboratory at a comfortable breeding environment temperature of 25°C, in the dark and quiet, and let them adapt for at least 1 hour.

[0022] At the beginning of the behavior, the temperature of the test device was set to 25°C, and each pup was gently taken out from the mother and placed in the test device, and its USVs and behavior video were recorded for 5 minutes. After the recording was completed, it was gently taken out of the test device and placed back to the mother, and the next pup was taken out until all the pups in the litter were recorded.

[0023] Ensure that each pup rests quietly for at least 40 minutes under the care of its mother during the two recordings. After a proper rest, the pups were placed in an experimental device set at 19°C to record USVs and behaviors during cold stimulation for 5 minutes. After the recording, the pups were allowed to return to room temperature and then placed back to their mothers.

[0024] USVs analysis software was used to identify USVs in the frequency range of 30-150kHz when the young mice were exposed to room temperature of 25℃ and cold stimulation of 19℃, and the USVs waveform was displayed in a visual form. By selecting each identified USVs wave, the number of USVs, average USVs duration, average USVs main frequency, average USVs frequency difference, and average USVs loudness indexes in the entire 5-minute recording time were automatically analyzed; comparison Shank3b + / + and Shank3b - / - Changes in the basic characteristic indicators of mouse USVs when the mouse pups are exposed to room temperature and cold stimulation. The purpose of mice sending out USVs is to attract the attention of the mother mouse so as to get adequate care. The greater the threat the mouse perceives to its environment, the more drastic the change in USVs. In a disease state, mice may show excessive sensitivity to the external environment. Conversely, in certain disease models, mice may show insensitivity to threatening stimuli, which may cause abnormal behavior of mice and even cause unexpected damage; Using Smart v3.0 behavioral video analysis software, by selecting the observation and calculation area and entering the actual size of the area, the software can automatically analyze according to the actual size of the device. By adjusting the contrast to increase the contrast between the mouse and the background, the shape of each frame of the mouse can be highlighted in real time, and the tracking mode can be set to "Center of Mass" to track the body position of the mouse pups in real time during movement. Through the automatic tracking and calculation of the software, the time, total distance, real-time speed, maximum speed and other indicators of the pups' movement at room temperature and cold stimulation can be compared. The status of the mouse pups can also be manually identified, such as recording the number of falls, the time of falls, the time of trembling, the time of lying on the wall, etc., to quantify the differences in the behavioral responses of the pups to different sensations.

[0025] The correlation analysis was performed on the mouse behavioral indicators and USVs indicators to study the correlation between the behavioral changes induced by temperature stimulation in neonatal mice. The test device included a temperature control unit, an ultrasonic detection unit, and a behavior detection unit; Temperature control unit: An aluminum plate and a controller that can freely adjust the temperature and maintain it within ±1°C of the set temperature were used; Ultrasonic detection unit: In a soundproof box, an ultrasonic probe was used to receive USVs signals, and the USVs signals were recorded by the USVs recording software in the computer. Behavior detection unit: In a soundproof box, an infrared night vision camera was used to record the behavior videos.

[0026] Combining the above data, the following results can be obtained: (1)Cold stimulation at 19°C Shank3b - / - could significantly increase the USVs in both the autism mouse model and wild-type neonatal mice, suggesting that both types of mice could perceive cold stimulation and express it through USVs; (2)The total movement of wild-type neonatal mice decreased significantly when exposed to cold stimulation, while the total movement time of autism mice remained unchanged, indicating their abnormal ability to perceive temperature; (3)Compared with room temperature, after cold stimulation, the duration of USVs in wild-type neonatal mice was significantly prolonged, the main frequency decreased, the frequency difference increased significantly, and the loudness increased significantly; (4) Shank3b - / - The changes in USVs signals in autism mice after cold stimulation below the comfortable temperature were inconsistent with those in wild-type mice, and showed insensitivity to cold stimulation, suggesting that they had obstacles in the understanding and expression of perception.

[0027] Refer to Figure 1 Figure A: The comparison of the number of USVs showed that Shank3b + / + the number of USVs in neonatal mice (N = 4 mice) tended to increase after cold stimulation at 19°C, Shank3b - / - the number of USVs in neonatal mice (N = 5 mice) increased significantly after cold stimulation at 19°C. B: The comparison of the total movement distance showed that Shank3b + / + the total movement distance of neonatal mice decreased significantly after cold stimulation, Shank3b - / - the total movement distance of neonatal mice did not change significantly after cold stimulation. C: The comparison of the duration of USVs showed that Shank3b + / + the duration of USVs in neonatal mice was significantly prolonged after cold stimulation, Shank3b - / - the duration of USVs in neonatal mice did not change significantly after cold stimulation. D: After cold stimulation Shank3b+ / + and Shank3b - / - There were no significant differences in the main frequency of USVs in mouse pups after cold stimulation. E: Comparison of USVs frequency differences showed that Shank3b + / + the frequency difference of USVs in mouse pups increased significantly after cold stimulation, Shank3b - / - the frequency difference of USVs in mouse pups did not change significantly after cold stimulation. F: Comparison of USVs loudness showed that Shank3b + / + the duration of USVs in mouse pups increased significantly after cold stimulation, Shank3b - / - the loudness of USVs in mouse pups did not change significantly after cold stimulation. Error bars indicate mean ±SEM. * P <0.05, *** P <0.001.

[0028] This method is mainly based on USVs to reflect the perception of temperature changes in mouse pups. Since the detection of USVs is not affected by changes in the experimental environment, this method can be further applied to evaluate various sensory abilities of mouse pups. For example, by changing the odor of the experimental environment and analyzing USVs, the olfactory perception ability of mouse pups can be evaluated; by applying different tactile stimuli to mouse pups and observing the changes in their USVs, the tactile perception ability can be evaluated, etc. In addition, this method also supports the evaluation of the effects of various compound sensory stimuli on juvenile mice, as much as possible to restore the situation where mice simultaneously perceive multiple factor stimuli in a complex environment, integrate various types of information, and reflect the entire peripheral and central functions by evaluating the final output USVs. Moreover, this method can be combined with methods for studying nerve functions, such as transgenic calcium indicator fiber recording methods, to jointly study its internal nerve mechanism.

[0029] Example 2 (1) Using this method, cold stimulation is applied to mouse pups and the changes in their perception ability are monitored. By recording and analyzing the frequency, duration, and pattern characteristics of ultrasonic vocalizations (USVs), the ability to process sensory information and the stress response of mouse pups in a cold environment can be accurately quantified. This USV-based behavioral detection method can not only reveal the abnormal integration and feedback mechanisms of autistic model mice to external temperature stimuli during the critical period of development, but also construct the correlation between sensory dysfunction and abnormal development of neural circuits by comparing the behavioral differences between typically developing individuals and disease models. The non-invasive feature of this method avoids the damage to animals caused by common methods (such as electrophysiology) and also minimizes the interference to animals as much as possible. Further, this technical framework can be extended to explore the neural mechanisms of the regulation of sensory input on social behavior and emotional state, and provide a standardized evaluation platform for analyzing the sensory abnormal phenotypes caused by anxiety behavior.

[0030] (2) Using this method, the functional maturation trajectory during the formation of the sensory and perceptual neural circuits in mouse pups can be dynamically traced, as well as the interaction mechanism between the sensory and perceptual and emotion-related neural circuits (the lack of feedback from the mother mouse when the mouse emits USVs may trigger anxiety emotions, etc.). Combining optogenetic or chemogenetic intervention means can accurately analyze the temporal characteristics of the high-level sensory integration center processing temperature information, and reveal the influence law of the critical period of synaptic plasticity regulation on the specialization of sensory and perceptual functions. By comparing the encoding differences of relevant neural circuits in mice between typical development and pathological models, the circuit nodes with abnormal temperature perception in neurodevelopmental disorders can be located. This research paradigm provides a cross-species theoretical model for simulating the sensory integration development in human infancy. The revealed regulation law of neural plasticity in the developmental window period may provide a mechanism-guided experimental framework for screening early intervention targets for sensory abnormalities in autism and other neurodevelopmental diseases.

[0031] (3) Using this method, by systematically clustering the USVs characteristic spectra of mouse pups with different neurodevelopmental disease models, a cross-disease phenotype database of sensory and perceptual function defects can be established. Combining single-cell transcriptome sequencing to reveal abnormal expression or assembly defects of sensitive molecules and locate the molecular targets with disordered sensory and perceptual information encoding. By using optogenetic or chemogenetic techniques to bidirectionally regulate specific circuit nodes, the key neural mechanisms of temperature perception signal transmission and their dynamic plasticity change laws can be verified. Further integrating calcium imaging technology to track the population neuron response patterns of key brain regions to cold stimuli can analyze the abnormal synchronization mechanism of the sensory-emotional integration network in developmental disorders. This multi-scale research strategy can also screen out potential drug molecules that regulate the excitability of primary sensory neurons. By establishing a USVs-behavior-molecule association prediction model using machine learning algorithms, it can provide theoretical support for the selection of personalized intervention timing. This systematic research framework will promote the innovation of the translational medicine research paradigm from phenotype-driven to mechanism-guided, and lay a cross-modal research foundation for the development of precision medical solutions for sensory hypersensitivity or hypoesthesia phenotypes.

[0032] The above content is only to illustrate the technical idea of the present invention, and the protection scope of the present invention cannot be limited thereby. Any modification made on the basis of the technical solution according to the technical idea proposed by the present invention falls within the protection scope of the claims of the present invention.

Claims

1. A method for quantitatively evaluating the sensory perception of a juvenile mouse model based on USVs, characterized in that, It includes the following steps: Set the temperature to the comfortable breeding environment temperature of the mice. Take each young mouse out from beside its mother and place it in the test device. Record its USVs and behavioral videos for a set duration. After the recording ends, take it out from the test device and put it back beside its mother, and then take out the next young mouse until all the young mice are recorded; After having a full rest beside its mother, place the cubs in the test device at a temperature lower than the comfortable temperature in turn, and record the USVs and behaviors for a set duration during cold stimulation. After that, let the cubs return to room temperature and then put them back beside their mother; Analyze the USVs characteristics of pups when receiving comfortable temperature and cold stimuli below the comfortable temperature, including: the number of USVs, the average duration of USVs, the average main frequency of USVs, the average frequency difference of USVs, and the average loudness index of USVs; compare Shank3b + / + and Shank3b - / - the changes in the USVs characteristic indexes of mouse pups at comfortable temperature and below the comfortable temperature, and obtain the behavioral phenotypes of infantile autism mice according to the changes Use Smart analysis software to analyze the recorded ethological movement videos and compare the differences in ethological indexes of the cubs at room temperature and during cold stimulation; Analyze the mouse ethological indexes and USVs characteristics to obtain the correlation between the behavioral changes induced by temperature stimulation in young mice; 2. The method for quantitatively evaluating the sensory perception of the juvenile mouse model based on USVs according to claim 1, wherein Each cub should be beside its mother for at least 40 minutes during the two test recordings; 3. The method for quantitatively evaluating the sensory perception of the juvenile mouse model based on USVs according to claim 1, wherein, The test device includes a soundproof box, in which a refrigeration plate, an ultrasonic probe and an infrared night vision camera are arranged; the ultrasonic probe and the infrared night vision camera are connected to a computer, and the refrigeration plate is a refrigeration plate with a temperature controller; 4. The method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs according to claim 1, wherein The comfortable temperature is 25 °C, and the cold stimulation temperature lower than the comfortable temperature is 19 °C; 5. The method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs according to claim 1, wherein The set duration is 5 minutes; 6. The method for quantitatively evaluating the sensory perception of a juvenile mouse model based on USVs according to claim 1, characterized in that, Before the test, place the mouse cage containing the young mice and their mother in an environment at the comfortable temperature and adapt in the dark and quiet for at least 1 hour; 7. The method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs according to claim 1, characterized in that, Cold stimuli below the comfortable temperature cause a significant increase in mouse USVs in both Shank3b autistic mouse models and wild-type mouse pups, suggesting that both types of mice can perceive cold stimuli and express them through USVs; The movement of wild-type mouse cubs significantly decreases during cold stimulation at a temperature lower than the comfortable temperature, while the total movement time of autistic mice remains unchanged, indicating its abnormal temperature perception ability; Compared with the comfortable temperature, after cold stimulation at a temperature lower than the comfortable temperature, the duration of USVs in wild-type mouse cubs is significantly prolonged, the main frequency is decreased, the frequency difference is significantly increased, and the loudness is significantly increased; Shank3b - / - After cold stimulation below the comfortable temperature, the changes in USVs signals in autistic mice were inconsistent with those in wild-type mice, and showed insensitivity to cold stimulation, suggesting that they have disorders in the understanding and expression of perception.

8. The method for quantitatively evaluating the sensory perception of a neonatal mouse model based on USVs according to claim 7, wherein The ethological indexes include the total movement time, total movement distance, number of falls and fall time; 9. Use of the method for quantitatively evaluating sensory perception of a neonatal mouse model based on USVs according to any one of claims 1-8, characterized in that, Conduct cold stimulation on mouse cubs to observe their perception ability, and use USVs to quantify the response of mouse cubs to cold stimulation at a temperature lower than the comfortable temperature, so as to evaluate the sensory perception ability of mouse models of neurodevelopmental disorders including autism in the early stage of the disease; 10. Use of the method for quantitatively evaluating sensory perception of a neonatal mouse model based on USVs according to any one of claims 1-8, characterized in that, Quantitatively compare the sensory perception abilities of mouse cubs of different disease models, and combine the molecular mechanism and circuit mechanism to assist in finding the intervention targets and treatment methods of the disease, providing methodological guidance for the research and development of therapeutic drugs and intervention methods for neurodevelopmental disorders including autism.

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