A method for constructing a mouse social novelty deficiency behavior model and application thereof

Abstract

The application discloses a kind of mouse social novelty defect behavior model construction method and application, belong to the technical field of disease animal model construction, the method with catechol as inducing agent induces mouse to produce social novelty behavior defect phenotype, the method simple and low in cost is operated, the mouse social novelty defect behavior model obtained has high specificity, i.e. only social novelty behavior defect can be induced without affecting mouse short-term memory, anxiety, depression, learning and memory and other behaviors, so as to accurately simulate the ASD social novelty defect induced by environmental factors, and provides reliable tool for the mechanism research and drug screening of ASD social novelty defect.

Description

Technical Field

[0001] This invention relates to the field of disease animal model construction technology, specifically to a method for constructing and applying a mouse social novelty deficiency behavior model. Background Technology

[0002] Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder that has become one of the most prevalent neurodevelopmental disorders worldwide. Its core symptoms include difficulties in social interaction and impaired language communication. Due to its complex etiology, there are currently no effective medications for treatment; symptom relief is achieved through rehabilitation therapies.

[0003] Social Novelty Deficit (SND) is a highly specific core social phenotype of ASD, distinct from ordinary "social avoidance / reduced social interest." SND refers to an individual's lack of normal exploratory motivation, preference, and approach behavior towards unfamiliar social objects or novel social interaction patterns in social situations. Simultaneously, the individual is unable to adjust their social behavior based on novel social cues, manifesting as a preference for familiar social objects, avoidance of unfamiliar social interactions, rigid and inflexible social behavior, and inability to adapt to new social scenarios. This phenotype is the core subtype of "persistent social interaction deficit" in DSM-5 and is one of the most typical social characteristics of human ASD patients. Social novelty deficiency is a hallmark social phenotype that distinguishes ASD from other neurodevelopmental disorders (ADHD, intellectual disability) and mental illnesses. However, current research on social novelty deficiency is extremely insufficient. Existing ASD models can only generalize and simulate abnormal social behavior, failing to accurately replicate the specific phenotype and pathological mechanisms of social novelty deficiency. Furthermore, the occurrence and development of this phenotype are directly linked to the gene-environment interaction etiology of ASD and damage to core neural circuits. Furthermore, besides autism, other mental illnesses such as schizophrenia, depression, and anxiety disorders are often accompanied by social dysfunction. For example, patients with depression may exhibit social withdrawal and a lack of interest in social activities, while patients with anxiety disorders are prone to fear and avoidance behaviors in social situations. These diseases also lack targeted treatment methods. Therefore, establishing reliable animal models of social deficits can provide experimental platforms for research on social impairments related to diseases such as ASD.

[0004] Currently, genetic modification and chemical drug induction methods are commonly used to construct models related to social deficits. Genetic modification, such as through... SHANK3 , MECP2Knocking out autism-sensitive genes to construct gene defect models is time-consuming and costly. Chemical induction methods, such as valproic acid (VPA) induction, require intraperitoneal injection of the drug into pregnant mice at specific times, which is difficult to control and may result in miscarriage of pregnant mice and death of newborn pups, affecting the number of viable individuals and also prolonging the modeling process. Therefore, establishing a rapid, simple, low-cost, and stable animal model of social deficits can significantly shorten the model construction cycle and improve the efficiency of scientific research, which is of great significance.

[0005] Catechol, chemically known as catechol or pyroquinone, with the molecular formula C6H4(OH)2, is widely distributed in various plants and their derivatives in nature. It is a colorless crystal that readily oxidizes and changes color upon exposure to light and air. It is soluble in approximately 2.3 times its volume of water and also soluble in organic solvents such as ethanol, ether, benzene, and chloroform. It is readily soluble in pyridine and strong alkaline solutions. As an important fine chemical raw material, catechin has wide applications in pesticides, pharmaceuticals, fragrances, dyes, photosensitive materials, and the rubber industry. It is commonly used as a rubber hardener, electroplating additive, skin disinfectant, hair dye, photographic developer, and antioxidant. Catechol exhibits strong biotoxicity; long-term or high-concentration exposure may affect the central nervous system, causing symptoms such as depression, convulsions, and respiratory failure. Currently, there are no reports of using catechin in establishing models related to social deficits. Summary of the Invention

[0006] In view of the technical problems existing in the background art, this invention proposes a new method for constructing a social novelty deficit model in mice by inducing it with drugs. The aim is to solve the problem that existing ASD models can only generalize and simulate social behavioral abnormalities and cannot accurately replicate the specific phenotype of social novelty deficit. This invention enables the rapid, simple and stable construction of a social novelty deficit behavioral model, providing a reliable tool for ASD-related mechanism research and drug screening.

[0007] To achieve the above objectives, the technical solution of the present invention is as follows: A method for constructing a mouse social novelty deficit behavior model, which uses catechol as an inducer to induce mice to specifically produce a social novelty deficit behavior phenotype.

[0008] Preferably, in the above construction method, the catechol is administered via subcutaneous injection. For example, in some embodiments of the present invention, a mouse model is prepared by intraperitoneal injection. Alternatively, the catechol can be prepared into a solution using physiological saline as a solvent and then administered subcutaneously.

[0009] Preferably, in the above construction method, the mice are acclimatized for ≥3 days before inducing the use of catechol.

[0010] Among the above construction methods, there are two construction strategies: Strategy 1: Administer an effective dose of catechol to mice once to establish an acute social novelty behavior deficit model; Strategy 2: Administer an effective dose of catechol to mice for n consecutive days, once a day, to construct a stable model of social novelty behavior deficit, where n is preferably 14-25.

[0011] In Strategy 1, mice acquire a specific phenotype of social novelty behavior deficit approximately 20 minutes after catechol administration, which can then be used for about 3 hours. In Strategy 2, extending the injection time within a certain period is beneficial for the retention time of the social novelty behavior deficit-specific phenotype; for example, in some embodiments of the present invention, after continuous administration to mice for 21 days, the specific phenotype of social novelty behavior deficit can be retained for at least 14 days. Furthermore, in Strategy 2, to avoid interruption of modeling due to infection, a rotation of injection sites can be used to address common complications of subcutaneous injection (such as abscesses, skin ulceration, etc.), reducing the incidence of complications and ensuring the continuity of the modeling cycle and animal welfare.

[0012] Preferably, in the above construction method, the effective dose of catechol is 25-50 mg / kg (catechol dosage / mouse body weight).

[0013] Preferably, the above construction method further includes the following steps: The three-box social experiment was used to verify the social novelty defect phenotype. The judgment criteria were: if the catechol-treated mice showed no significant difference in exploration time between familiar mice and unfamiliar mice, while the control mice took significantly longer to explore unfamiliar mice, the model was considered successfully constructed; or / and, At least one of the following tests was used for exclusion: novel object recognition test, open field test, elevated cross maze test, tail suspension test, forced swimming test, marble burial test, T-maze test, and conditioned fear test, to verify that the mice had no abnormalities in short-term memory, anxiety, or depression, and to ensure that the phenotype was a social novelty-specific defect.

[0014] The mouse social novelty deficiency behavior model constructed in this invention can be used for at least the following: 1) Screening or evaluating drugs that intervene in social deficits of ASD; 2) Investigate the pathogenesis of ASD, especially the neural circuits and molecular mechanisms related to social deficits; 3) Screening for diagnostic biomarkers associated with ASD’s social novelty deficit.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention provides a novel method for establishing a mouse social novelty deficiency behavior model induced by catechol. This method does not require gene editing equipment and uses inexpensive catechol raw materials, which significantly reduces the cost of model preparation. Moreover, it has high construction efficiency, and can quickly establish acute social novelty deficiency behavior models as well as stable social novelty deficiency behavior models (e.g., a model whose behavioral phenotype can be maintained for more than 14 days can be obtained in only 21 days, which is much shorter than the time required to establish a genetic mutation model).

[0016] The mouse social novelty deficiency behavioral model obtained by this invention is highly specific, inducing only social novelty deficiency without affecting other behaviors such as short-term memory, anxiety, depression, and learning memory in mice. This avoids interference from non-target phenotypes in experimental results, and thus accurately simulates ASD social novelty deficiency induced by environmental factors, providing a reliable tool for related mechanism research and drug screening. Furthermore, the mouse social novelty deficiency behavioral model provided by this invention can simulate the pathological processes mediated by environmental factors (such as metabolic abnormalities and exogenous toxin exposure) in human ASD, effectively compensating for the inability of genetic models to reproduce environmental pathogenesis.

[0017] The model construction method provided by this invention is highly operable: it uses intraperitoneal injection for drug administration, is simple to operate, does not require complex surgery or microinjection techniques, and can be quickly mastered by those skilled in the art. Attached Figure Description

[0018] To more clearly illustrate the technical solution of the present invention, the accompanying drawings used in the present invention will be briefly described below. Obviously, the drawings described below are merely some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0019] Figure 1 This is a flowchart illustrating the construction of a mouse model of social novelty deficiency behavior induced by catechols in this invention. Figure 2 This is a graph showing the results of mouse mortality under acute exposure to different doses of catechol in Example 1; Figure 3 The diagram shows the results of the open field experiment in mice under acute exposure to different doses of catechol in Example 1. A is a schematic diagram of the open field experiment, B is a comparison of the total movement distance of the mice, and C is a comparison of the movement distance of the central region of the mice. Figure 4 The diagram shows the results of the three-box social experiment in mice under acute exposure to different doses of catechol in Example 2. A is a diagram of the three-box social experiment, B is a comparison of the exploration time in the habituation stage, C is a comparison of the exploration time in the social ability test stage, and D is a comparison of the exploration time in the social novelty test stage. Figure 5The figures show the results of the new object recognition experiment in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the new object recognition experiment, B is the comparison result of the exploration time in the first stage, and C is the comparison result of the exploration time in the second stage. Figure 6 The figures show the results of the elevated cross maze experiment in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the elevated cross maze experiment, B is the comparison result of open-arm dwell time, and C is the comparison result of closed-arm dwell time. Figure 7 Figure A shows the results of the marble burial experiment in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the marble burial experiment, and B is a comparison of the number of buried marble balls. Figure 8 Figure A shows the results of the T-maze test in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the T-maze test, and B is the result of the accuracy comparison. Figure 9 The diagram shows the results of the forced swimming experiment in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the forced swimming experiment, and B is a comparison of the proportion of immobility time. Figure 10 The following figures show the results of the tail suspension test in mice under acute exposure to different doses of catechol in Example 2. A is a schematic diagram of the tail suspension test, and B is a comparison of the proportion of immobility time. Figure 11 The figure shows the results of the three-box social experiment in mice under acute catechol exposure in Example 3. A is the comparison of exploration time in the habituation stage, B is the comparison of exploration time in the social ability test stage, and C is the comparison of exploration time in the social novelty test stage. Figure 12 The figures show the results of the new object recognition experiment in mice under acute catechol exposure in Example 3. A is a comparison of the exploration time of the two objects during the habituation stage, and B is a comparison of the exploration time of the two objects during the new object recognition stage. Figure 13 The diagram shows the results of the open field experiment on mice under acute exposure to catechol in Example 3. A represents the total distance traveled by the mice, and B represents the distance traveled by the mice in the central region. Figure 14 The image shows the results of the elevated cross maze experiment in mice under acute catechol exposure in Example 3. A represents the distance traveled with the arms open, and B represents the time spent with the arms open. Figure 15 This is a diagram showing the results of the marble burial experiment on mice under acute exposure to catechols in Example 3; Figure 16 This is a diagram showing the results of the forced swimming experiment in mice under acute exposure to catechols in Example 3; Figure 17This is a diagram showing the results of the tail suspension experiment in mice under acute exposure to catechols in Example 3; Figure 18 The graph shows the results of detecting the maintenance of social novelty behavior deficit in mice after acute exposure to catechol in Example 3, one week and two weeks after modeling. A is the detection result one week after modeling, and B is the detection result two weeks after modeling. Explanation of reference numerals in the attached figures: P ≤0.0001, P ≤0.001, P ≤0.01, P ≤0.05, ns indicates no significance. Detailed Implementation

[0020] The technical solution of the present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0021] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains; the terminology used herein is for the purpose of describing particular implementations only and is not intended to limit the invention.

[0022] Currently, most widely used animal models of autism spectrum disorder rely on gene editing technology, which is time-consuming, costly, and difficult to simulate social behavioral deficits induced by environmental factors. Although some studies have attempted to induce behavioral abnormalities using chemicals such as valproic acid, the induction period is long, phenotypic stability is poor, and non-target behavioral interference is often present, limiting their application in mechanism research and drug screening. Addressing the problems of complex ASD model construction and the inability to accurately replicate the specific phenotype of social novelty deficit in existing technologies, this invention utilizes catechols to establish a novel method for preparing a mouse model of social novelty deficit behavior. This model preparation method is simple and highly specific, inducing only social novelty deficit behavior without affecting other behaviors such as short-term memory, anxiety, depression, and learning memory in mice. Its scientific, academic, and translational value far exceeds that of generalized social deficit models, and it represents a breakthrough in solving the current problems of heterogeneity in ASD social phenotypes, ambiguity in mechanisms, and inefficient translation.

[0023] like Figure 1 As shown, the method for constructing a mouse social novelty deficiency behavior model provided in this embodiment of the invention includes the following steps: S1. The mice were acclimatized for no less than 3 days. S2. Prepare a catechol solution using physiological saline as a solvent; S3. The catechol solution obtained in step S2 is administered subcutaneously to the mice obtained in step S1, wherein the injection method is as follows: A single effective dose of catechol was administered to mice to establish an acute social novelty behavior deficit model; or, Mice were given an effective dose of catechol once a day for n consecutive days to construct a stable model of social novelty behavior deficit, where n was selected from 14-25.

[0024] S4. Verify the social novelty defect phenotype using a three-box social experiment. If the catechol-treated mice do not show a significant difference in exploration time between familiar mice and unfamiliar mice, while the control mice (injected with the same volume of saline) take significantly longer to explore unfamiliar mice, then the model is considered to have been successfully constructed.

[0025] Furthermore, in some embodiments of the present invention, the mice are male C57BL / 6J mice, and the acclimatization conditions include: 12 hours of light... Dark cycle, temperature 20 22℃, humidity 45% 60%.

[0026] Furthermore, in some embodiments of the present invention, the effective dose of catechin is 25-75 mg / kg.

[0027] Furthermore, in some embodiments of the present invention, The three-box social experiment, as a unique behavioral testing method, has become an important tool in animal behavior and neuroscience research; especially in animal models of ASD and other social disorders, this experiment provides a platform for in-depth understanding of social behavior and cognitive processes. The three-box social experiment consists of the following three phases: Habituation phase: Place the mice in the central area and allow them to explore freely for a period of time, and record the results; Social skills test: A strange mouse (restrained in a cylindrical cage, denoted as S1) was placed in one area, and an empty restrained cage (denoted as S2) was placed in the other area. The mouse's exploratory behavior was recorded. Social novelty test: Keep the familiar S1, replace the empty cage on the other side with another new unfamiliar mouse (denoted as new S2), and record the mouse's exploration behavior for 10 minutes again; The behavioral analysis software (such as VisuTrack) was used to automatically record the exploration time of mice for each target within the range of the restraint cage. The judgment criteria were: if there was no statistically significant difference in the exploration time of S1 and new S2 in the drug administration group during the social novelty test (p>0.05), while the exploration time of new S2 in the control group was significantly longer than that of S1 (p<0.05), then the social novelty behavior deficit model was judged to have been successfully constructed.

[0028] Specifically, in an embodiment of the present invention, the device for the three-box social experiment is a 90×30×30 cm acrylic three-box, divided into three areas: left, middle, and right. Cylindrical restraint cages are placed in the two side areas, and the exploration range is defined as 7 cm around the restraint cages.

[0029] Furthermore, in some embodiments of the present invention, the construction method further includes the following steps: S5. Use at least one of the following experiments to eliminate interfering factors: novel object recognition experiment, open field experiment, elevated cross maze experiment, tail suspension experiment, forced swimming experiment, marble burial experiment, T-maze experiment, and conditioned fear experiment; if there is no significant difference between catechol-treated mice and control mice in these experimental tests, it can be further proved that the induced defect is a social novelty-specific defect.

[0030] The novel object recognition experiment can assess the short-term memory and cognitive level of mice. This experiment consists of two phases: a training phase to train mice to remember objects, and a testing phase to observe and record the time mice spend exploring familiar and new objects. Mice with normal short-term memory will spend more time exploring new objects. Objects the size of a mouse or slightly larger can be used to stimulate the mice's exploration desire. To reduce bias, mice should be able to climb two objects simultaneously, although climbing ability may increase interest in exploration. Therefore, the time spent sitting on one object is not counted in the exploration time. The two objects are made of durable materials and have distinctly different shapes to avoid damage during the experiment and to prevent potential harm or stimulation to the animals. Overly simple objects are not used to stimulate the mice's exploration desire. The experiment can be conducted in a 45×45×45 cm open field apparatus, with the two objects placed side-by-side 15 cm from the wall, 15 cm apart. Low light intensity is used. After each phase, the apparatus is wiped with 75% alcohol to remove odor. The exploration time and movement trajectory of the mice can be recorded using VisuTrack software.

[0031] The open field test and the elevated cross maze test can be used to assess anxiety levels, while the tail suspension test and the forced swimming test can be used to assess depressive states. The T-maze self-alternation test and the conditioned fear test can be used to verify spatial working memory and learning memory.

[0032] The following are some specific embodiments. It should be noted that the embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in this field or according to the product instructions. Reagents or instruments used, unless otherwise specified, are all conventional products that can be obtained commercially.

[0033] Example 1 This example determined the effect of different concentrations of catechol solutions on the survival status of mice, specifically including the following procedures: Forty healthy male C57BL / 6J mice (weighing 20±2 g) aged 8 weeks were selected and housed in an SPF-grade animal facility under the following environmental conditions: 12 h light... Dark Loop (07:00) (19:00 light), temperature 22±1℃, humidity 50±5%, free access to food and water. After 7 days of acclimatization, the animals were used in experiments.

[0034] Catechol powder (purity ≥ 99%) was prepared into catechin solutions of different concentrations with sterile physiological saline and used immediately after preparation.

[0035] Mice were randomly divided into 5 groups (n=8): a normal saline control group (NS) and groups treated with 25 mg / kg, 50 mg / kg, 75 mg / kg, and 100 mg / kg catechol. Mice in each group were induced to mate at 9:00 AM. At 10:00, mice were given an intraperitoneal injection of different doses of catechol, with the injection volume calculated as 0.0075 ml / g body weight. The mice were immediately returned to their original cages after the injection and observed for 72 hours. The survival status, behavioral status, and changes in food and water intake were recorded.

[0036] The results are as follows Figure 2 As shown, mice in the 100 mg / kg catechol administration group showed improvement 2 days after administration. All mice died within 4 hours. The group treated with 75 mg / kg catechol showed acute toxic reactions such as convulsions and rapid breathing. Mice in the other groups survived and no obvious abnormal behavior was observed.

[0037] Then, open field experiments were conducted on mice in the 25 mg / kg, 50 mg / kg, and 75 mg / kg catechol administration groups and the control group. Specifically, a 45×45×45 cm open field box was used, and the time spent in the central area and the total distance traveled within 10 minutes were recorded. The entire experiment was automatically recorded using an animal behavior analysis system.

[0038] The results of the open field experiments for each group are as follows: Figure 3 As shown in BC, the 75 mg / kg catechol administration group showed impairment in total motor function and a significant reduction in exploration of the central region, indicating that high-dose catechol exposure caused significant damage to mouse behavior. Therefore, the high-dose group data were not considered in subsequent validation experiments.

[0039] Therefore, based on the above results and animal welfare considerations, 25 mg / kg and 50 mg / kg can be used as effective and safe concentrations for subsequent model construction, with 50 mg / kg being the preferred choice due to its phenotypic stability and lack of acute toxicity.

[0040] Example 2 This example established an acute model using catechol induction, specifically including the following procedures: Eight-week-old male C57BL / 6J mice (weighing 20±1 g) were acclimatized for 3 days. They were randomly divided into four groups: a control group (NS) receiving an equal volume of physiological saline intraperitoneally; a low-dose group (CAT) receiving a normal saline solution; and a control group receiving an equal volume of normal saline intraperitoneally. 25), intraperitoneal injection of 25 mg / kg catechol solution, preferred dose group (CAT). 50), intraperitoneal injection of 50 mg / kg catechol solution, high-dose group (CAT) 75), mice were injected intraperitoneally with 75 mg / kg catechol solution. The injection volume was 0.0075 mL / g, administered 20 min before the behavioral test. The housing environment for each group of mice was the same as in Example 1.

[0041] The following behavioral tests were performed on the mice in each group: (1) Three-box social experiment.

[0042] A 90×30×30 cm acrylic three-box assembly (divided into left, middle, and right areas, such as...) is used. Figure 4 As shown in A), according to "habituation" Social skills test The "Social Novelty Test" was conducted in three phases: Habituation phase: Place the mouse in the central area and allow it to explore freely for 10 minutes; Social skills test: An unfamiliar mouse (restrained in a cylindrical cage, denoted as S1) was placed in one area, and an empty restraint cage (denoted as S2) was placed in the other area. The mouse was tested for 10 minutes. Social novelty test: The familiar S1 is retained, and the other empty cage is replaced with another new unfamiliar mouse (denoted as new S2). The test mice are tested for 10 minutes. After each stage, the device was wiped with 75% alcohol to remove the odor. The exploration time and movement trajectory of the mice within a 7cm range of the restraint cage were automatically recorded using a behavior analysis system.

[0043] The criteria for determining the social novelty of the three boxes mentioned above are: if there is no statistically significant difference in the exploration time of S1 and the new S2 among the mice in the drug-treated group during the social novelty test phase ( P >0.05), while the control group took significantly longer to explore the new S2 than S1 ( PIf the value is less than 0.05, the social novelty behavior defect model is considered to have been successfully constructed.

[0044] The results of the three-box socialization experiment for each group of mice in this example are as follows: Figure 4 As shown in BD: the exploration time for the new S2 in the NS group was significantly longer than that in S1. P <0.001), while CAT 25 groups, CAT 50 sets and CAT There was no significant difference in the exploration time between S1 and the new S2 among the 75 groups. P >0.05). This result indicates that a single intraperitoneal injection of catechol at concentrations of 25 mg / kg, 50 mg / kg, and 75 mg / kg can stably induce social novelty deficits in mice.

[0045] (2) New object recognition experiment.

[0046] Since cognitive deficits in mice can severely interfere with the assessment of social novelty deficits, we conducted a new object recognition experiment on mice after acute modeling to determine that their cognitive and short-term memory abilities had not changed.

[0047] The results are as follows Figure 5 As shown: The novel object recognition stage in mice ( Figure 5 B) No abnormalities were observed. This indicates that the catechol-induced social novelty deficit model is not due to short-term memory impairment or cognitive decline.

[0048] (3) Verification of behavioral phenotype specificity.

[0049] To verify the specificity of the social novelty behavior deficit model obtained by the method of this invention, this example tested whether the social novelty behavior deficit model would cause anxiety-like behavior, spatial memory deficit, and depression-like behavior in mice through the following experiments: A new batch of mice was selected, and the CAT assay constructed in Example 2 was followed. The modeling methods for 50 groups (n=10) and the NS group (n=10) were followed by the following behavioral tests 20 minutes after modeling (test interval ≥ 1 hour to reduce the impact of fatigue): ① Elevated cross maze experiment: Record the proportion of time spent with arms open within 5 minutes; ② Marble burying experiment: The number of marbles buried by mice within 30 minutes was recorded to assess anxiety levels; ③ Tail suspension test: Record the cumulative immobility time of the mouse within 6 minutes; ④ Forced swimming test: Record the cumulative floating time of mice in the pool within 6 minutes to assess depression level; ⑤ T-maze autonomous alternation experiment: The correct alternation rate in 10 tests was recorded to assess the spatial memory level of mice; The results are as follows Figures 6-10 As shown: CAT There were no statistically significant differences between the 50 group and the NS group in all the above tests (p>0.05), meaning that the model showed no difference in irrational behavior, spatial memory deficits, and depressive-like behavior between the two groups. This indicates that the catechol-induced behavioral deficits are highly specific and do not affect the mice's short-term memory, anxiety level, depressive-like behavior, or spatial working memory.

[0050] Example 3 This example demonstrates the establishment of a stable model for maintaining a social novelty deficit phenotype for a longer period using catechol induction. The specific steps include the following: Eight-week-old male C57BL / 6J mice (weighing 20±1 g) were used for acclimatization for 3 days. The rearing conditions included a 12-hour light-dark cycle (07:00-19:00), a temperature of 20-22℃, and a humidity of 45-60%, with 3-5 mice per cage.

[0051] The catechin solution was prepared according to Example 1.

[0052] After acclimatization, mice were randomly divided into a control group (NS, n=9) and a catechol exposure group (CAT, n=10). For 21 consecutive days at 10:00 AM, the CAT group was injected intraperitoneally with catechol solution (daily dose of 50 mg / kg), while the NS group was injected intraperitoneally with an equal volume of physiological saline. After injection, the mice were returned to their cages and fed normally.

[0053] Following 21 days of continuous catechol exposure, the mice in each group underwent the following behavioral tests on day 22 (completed between days 22 and 28, with no prior catechol exposure in the experimental groups before each behavioral test): (1) Three-box social experiment.

[0054] The test procedure is as described in Example 2. The results are as follows: Figure 11 As shown, 21 days of continuous exposure to catechols induced a deficit in social novelty behavior in mice, indicating successful modeling.

[0055] (2) New object recognition experiment.

[0056] Referring to Example 2, the cognitive abilities and short-term memory levels of each group of mice were assessed through a novel object recognition experiment.

[0057] Figure 12 The results of the new object recognition experiment showed that there was no significant difference between the mice and the control group in terms of short-term memory and cognitive level regarding new objects.

[0058] (3) Verification of behavioral phenotype specificity.

[0059] Referring to Example 2, the motor ability, anxiety-like behavior, and depression-like behavior of the mice after modeling were evaluated.

[0060] Figures 13-17 The results of the open field test, the elevated cross maze test, the marble burial test, the forced swimming test, and the tail suspension test were respectively. These results showed that there were no significant differences between the experimental group mice and the control group mice.

[0061] To further test the stability of the model obtained in this invention, a three-box social experiment was conducted one week after the model was established to evaluate the social novelty behavior deficit in mice. The results are as follows: Figure 18 As shown in Figure A, the mice maintained the social novelty deficit phenotype one week after modeling. Then, two weeks after modeling, a three-box social experiment was conducted again, with the following results: Figure 18 As shown in B, the social novelty behavior deficit model remains stable. These results demonstrate that the model established in this invention not only possesses the advantages of low cost and high specificity, but also exhibits a high level of model stability.

[0062] In summary, this invention can rapidly establish both an acute and stable social novelty deficiency behavioral model using catechol induction. Moreover, the resulting mouse social novelty deficiency behavioral model is highly specific and can accurately simulate ASD social novelty deficiency induced by environmental factors, showing potential for application in ASD-related mechanism research and intervention drug screening.

[0063] It should be noted that the present invention is not limited to the above-described embodiments. The above embodiments are merely examples, and any embodiments that have the same structure and perform the same effects as the technical concept within the scope of the present invention are included within the scope of the present invention. Furthermore, various modifications that can be conceived by those skilled in the art to the embodiments, and other ways of constructing by combining some of the constituent elements of the embodiments, without departing from the spirit of the present invention, are also included within the scope of the present invention.

Claims

1. A method for constructing a mouse social novelty deficit behavior model, characterized in that, Catechol was used as an inducer to induce a social novelty deficit phenotype in mice, with an effective dose of 25-50 mg / kg.

2. The construction method according to claim 1, characterized in that, The catechol is administered via intraperitoneal injection.

3. The construction method according to claim 1, characterized in that, Mice were acclimatized for ≥3 days before inducing the use of catechols.

4. The construction method according to claim 1, characterized in that, Includes the following steps: A single effective dose of catechol was administered to mice to establish an acute social novelty behavior deficit model.

5. The construction method according to claim 1, characterized in that, Includes the following steps: A stable model of social novelty behavior deficit was constructed by administering an effective dose of catechol to mice once a day for n consecutive days, where n was selected from 14-25.

6. The construction method according to claim 4 or 5, characterized in that, It also includes the following steps: The three-box social experiment was used to verify the social novelty defect phenotype.

7. The construction method according to claim 6, characterized in that, It also includes the following steps: Elimination was carried out using at least one of the following: new object recognition experiment, open field experiment, elevated cross maze experiment, suspended tail experiment, forced swimming experiment, marble burial experiment, T-maze experiment, and conditioned fear experiment.

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