Application of mouse model in evaluating intelligence level change
阅读说明：本技术 小鼠模型在评估智力水平变化中的应用 (Application of mouse model in evaluating intelligence level change ) 是由 琚存祥 赵静 王晶晶 陈中 于 2020-08-03 设计创作，主要内容包括：本申请涉及一种小鼠模型在评估智力水平变化中的应用,所述小鼠模型为高智力水平小鼠模型,所述高智力水平小鼠模型中的小鼠的染色体至少包含野生型小鼠来源的1号染色体的全部或部分,所述评估包含实施选自以下组的试验：开场试验、糖水偏好试验、水迷宫试验、空间探查Y迷宫试验、主动回避Y迷宫试验和疲劳悬转棒试验。(The present application relates to the use of a mouse model for assessing a change in mental level, said mouse model being a high mental level mouse model in which the chromosomes of a mouse comprise at least all or part of a wild type mouse-derived chromosome 1, said assessment comprising performing an assay selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.)
1. Use of a mouse model for assessing a change in mental level, the mouse model being a high mental level mouse model in which the mouse chromosome comprises at least all or part of chromosome 1 from a wild type mouse, the assessment comprising performing an assay selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.
2. The use of claim 1, wherein the mouse is derived from strain C57 BL/6J.
3. Use according to claim 1, wherein said mental level is determined by evaluating one or more conditions of the group consisting of: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
4. Use according to claim 3, wherein the motor ability is assessed by the open field test.
5. Use according to claim 3, wherein the emotional state is assessed by the sugar water preference test.
6. Use according to claim 3, wherein the learning ability and/or the memory ability is assessed by the water maze test.
7. Use according to claim 3, wherein said memory capacity and/or said spatial exploration capacity is evaluated by said spatial exploration Y maze test.
8. The use of claim 3, wherein the memory capacity is assessed by the active avoidance Y maze test.
9. Use according to claim 3, wherein the motor ability is assessed by the fatigue swing rod test.
10. A system comprising a high intelligent level mouse model in which the chromosomes of a mouse comprise at least all or part of wild type mouse derived chromosome 1, the system further comprising a discrimination module that assesses changes in the intelligent level of a mouse in the high intelligent level mouse model, the discrimination module comprising reagents and/or instrumentation required to carry out a test selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.
Technical Field
The application relates to the field of biomedicine, in particular to application of a mouse model in assessing intelligence level change.
Background
The Alzheimer's Disease (AD) experimental animal model is used as an important tool for researching pathogenesis, early diagnosis and drug treatment of the disease, and the scientificity, accuracy and diversity of the model are important factors for reducing clinical failure of the drug in the III stage. The introduction of variant genes related to Alzheimer's disease into animal gene system is used to simulate the pathological features and behavior change of Alzheimer's disease, and is the starting point of the research of Alzheimer's disease experimental animal model.
However, the above models all have late onset, and false positive or false negative results are likely to occur in the detection of the effect of the drug in the clinic. This is in direct contrast to the characteristic of Alzheimer's disease that its phenotypic characters are complex and affected by polygene, environment and age. At present, no suitable animal resource is found as a research tool, which is the pain point of the research and development of new Alzheimer disease medicines.
Disclosure of Invention
The application provides an application of a mouse model in evaluating the change of intelligence level. The high mental level mouse model described herein may have at least one beneficial effect selected from the group consisting of: 1) the learning ability is high; 2) has higher memory capacity; 3) the genetic diversity is possessed; 4) facilitates screening and/or manufacture of a medicament for treating a disease associated with mental levels (e.g., Alzheimer's disease); 5) advantageously improve the safety and/or effectiveness of a medicament and/or treatment regimen for treating a condition associated with mental performance (e.g., Alzheimer's disease); and 6) molecular mechanisms that facilitate comprehensive analysis of mental-level related disorders.
In one aspect, the present application provides the use of a mouse model for assessing a change in mental level, the mouse model being a high mental level mouse model in which the mouse chromosome comprises at least all or part of a wild type mouse-derived
In certain embodiments, the mouse is derived from strain C57 BL/6J.
In certain embodiments, the mental level is determined by evaluating one or more conditions of the group consisting of: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
In certain embodiments, the exercise capacity is evaluated by the open field test.
In certain embodiments, the emotional state is assessed by the sugar water preference test.
In certain embodiments, the learning ability and/or the memory ability is assessed by the water maze test.
In certain embodiments, the memory capacity and/or the spatial exploration capacity is assessed by the spatial exploration Y maze test.
In certain embodiments, the memory capacity is evaluated by the active avoidance Y maze test.
In certain embodiments, the exercise capacity is evaluated by the fatigue swing rod test.
In another aspect, the present application provides a system comprising a high intelligent level mouse model in which the chromosomes of a mouse comprise at least all or part of wild type mouse-derived
Other aspects and advantages of the present application will be readily apparent to those skilled in the art from the following detailed description. Only exemplary embodiments of the present application have been shown and described in the following detailed description. As those skilled in the art will recognize, the disclosure of the present application enables those skilled in the art to make changes to the specific embodiments disclosed without departing from the spirit and scope of the invention as it is directed to the present application. Accordingly, the descriptions in the drawings and the specification of the present application are illustrative only and not limiting.
Drawings
The specific features of the invention to which this application relates are set forth in the appended claims. The features and advantages of the invention to which this application relates will be better understood by reference to the exemplary embodiments described in detail below and the accompanying drawings. The brief description of the drawings is as follows:
FIGS. 1-4 show the results of an open field test of high mental level mice as described herein.
FIG. 5 shows the results of a sugar water preference test of high intellectual level mice as described herein.
FIGS. 6-8 show the results of a water maze test in high mental level mice as described herein.
FIGS. 9-10 show the results of a spatially exploratory Y maze test for high mental level mice as described herein.
FIGS. 11-12 show the results of an active avoidance Y maze test in high mental level mice as described herein.
FIG. 13 shows the results of a fatigue hanging rod test of high mental level mice as described herein.
FIG. 14 shows the results of body weight measurements of mental level mice as described herein.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification.
Definition of terms
In the present application, the term "mouse model" generally refers to an animal model that may be used, which may include mice (cMus musculus) But may also include cells, tissues or other materials derived therefrom.
In the present application, the term "wild-type mouse" generally refers to a wild-type mice (WT), which possesses a phenotype of a genetic characteristic in the natural population, considered to be of the "normal" (normal) type. Similarly, alleles of a particular gene conferring this phenotype are considered to be a "normal" type in the natural population. For example, the wild-type mice may include mice grown in a natural state captured from various places.
In this application, the term "high intellectual level" generally means that the intellectual level is significantly increased compared to the usual situation. The mental level may be measured by the wecker adult mental scale, for example, a score greater than 120 may be considered a high mental level. The mental level of a mouse may also be measured experimentally, for example, by an open field experiment, a sugar water preference experiment, a water maze assessment experiment, a spatially exploratory Y maze experiment, and/or an active avoidance Y maze experiment. Other methods of measuring the level of mental capacity in mice are also known to those skilled in the art, for example reference may be made to Matzel, L.M. et al, Indvidual differences in the expression of a 'general' learning ability in the microorganism, journal of Neuroscience, 23, 6423-.
In the present application, the term "all or part of a chromosome" generally refers to the complete chromosome or a fragment thereof. The chromosome may refer to a structure unique to eukaryotes, which is composed of double-stranded helical deoxyribonucleic acid and histone. The chromosome may be an aggregate formed by deep compaction of genetic material with genetic properties within the cell. At interphase, the chromosomes may exist in the form of chromatin. The mouse may have 20 pairs of chromosomes.
In the present application, the term "replacement" generally refers to the replacement of a chromosome in a mouse with the complete chromosome of wild-type mouse origin or a fragment thereof. For example, the replacement may be such that the mouse comprises the entire chromosome or fragment thereof of wild type mouse origin. On this basis, the replacement may also be such that the corresponding proto-chromosome in the mouse is replaced, and/or lost function. Such alternative methods are known to those skilled in the art and may be accomplished, for example, by hybridization and/or gene editing.
In the present application, the term "gene editing" generally refers to a technique of DNA insertion, deletion, modification or substitution in a genome. The gene editing may utilize Zinc Finger Nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), or CRISPR-Cas9 systems. For example, the gene editing may include a knock-in method. The method of knock-in can be a method commonly used by those skilled in the art, for example, see "gene targeting: one practical method ", Joyner editors, Oxford university Press Inc., 2000.
In the present application, the term "specific SNP site" generally refers to a SNP site specific for a certain genotype. The SNP site may be a position of a nucleic acid exhibiting a Single Nucleotide Polymorphism (Single Nucleotide Polymorphism). In this application, a SNP may refer to a change in the DNA sequence caused by a change in a single nucleotide-A, T, C or G. A typical SNP site may be two alleles. The SNP sites may be located in non-coding regions, and/or coding regions. The detection method of the SNP site may be selected from the group consisting of: DNA sequencing, single-strand conformation polymorphism (SSP), electrochemical analysis, denaturing HPLC and gel electrophoresis, restriction fragment length polymorphism and hybridization analysis.
In the present application, the term "primary cell culture" generally refers to the product obtained by culturing primary cells. The primary cells may be derived from excised animal tissue. Isolated primary cells may include adherent cells that require attachment for growth, as well as suspension cells that can grow without attachment. The primary cell culture may be obtained by culturing under in vitro conditions after dissociation of the primary cells, e.g. by enzymatic treatment. The primary cell culture can be converted to an unlimited number of subcultures.
In this application, the term "athletic ability" generally refers to the ability to participate in sports and training. The exercise capacity may include aerobic exercise capacity, muscular strength, body flexibility, balance capacity, and reaction capacity. The motor ability can be a comprehensive expression of a plurality of factors such as physical form, quality, skill and mental ability.
In the present application, the term "emotional state" generally refers to a psycho-physiological structure (psycho-physiological structures) that includes cognition, mood, and emotion. Emotional states can vary along three main aspects: titer (value), arousal (arousal) and Motivational intensity (kinetic science) (see Current directives in Psychological science, 2013-8, 22 (4): 301-. In the present application, the emotional state may include features of depression, caution, and the like.
In this application, the term "learning ability" generally refers to the psychological characteristics that an individual needs to possess in order to engage in learning activities. The learning capabilities may include imagination, attention, perceptual observation capabilities, reading capabilities, analysis capabilities, operational capabilities, adaptation capabilities, inductive summary capabilities, problem solving capabilities, or a combination thereof.
In this application, the term "memory ability" generally refers to the ability to remember, maintain, recognizably and reproduce the content and experience reflected by objective things. The memory capacity may include sensory memory capacity, short-term memory capacity, and long-term memory capacity.
In this application, the term "spatial exploration capability" generally refers to the ability to explore the shape and/or location of an object. The spatial exploration capability includes observing, thinking, imagining, recognizing, and/or fumbling about the shape and/or position of an object.
In the present application, the term "Alzheimer's disease" generally refers to presenile dementia, senile dementia, a neurodegenerative disease with a slow progression and a progressive deterioration over time. The most common early symptoms are loss of short-term memory (difficulty in remembering what happened recently), and as the disease progresses, at least one of the following symptoms may develop: language disorders, disorientation (e.g., easy getting lost), emotional instability, loss of motivation, inability to self-care, and behavioral problems. The true cause of alzheimer's disease is still unknown to date, and its progression is associated with fibrous amyloid plaque deposition and Tau protein in the brain. There is currently no treatment that can prevent or reverse the course of the disease, and only a few methods may temporarily alleviate or ameliorate the symptoms.
In the present application, the term "chronic encephalopathy syndrome" generally refers to an organic syndrome transformed from a slowly progressing encephalopathy or acute encephalopathy syndrome. The clinical manifestations are mainly dementia, personality changes and amnestic syndrome without accompanying disturbance of consciousness. Can be accompanied by chronic psychotic symptoms such as depression, mania-like or schizophrenia-like manifestations. The chronic encephalopathy syndrome may be caused by chronic organic disease, or may be seen as sequelae caused by the persistence of acute encephalopathy syndrome.
In the present application, the term "Attention Deficit Disorder" refers generally to Attention Deficit Disorder, ADD or Attention Deficit hyperactivity Disorder, ADHD, which may include Attention Deficit Disorder in adults and/or Attention Deficit Disorder in children. The characteristics of the attention deficit disorder may be inattentive, overactive, doing without regard to consequences, and the like. In addition to this, there is an unaged behavior and individuals with attention deficit may also have problems with mood that is difficult to regulate. The attention deficit disorder may be caused by the interaction of factors such as genes, environment, and society.
In the present application, the term "childhood autism" generally refers to childhood-autism, which originates before the age of 3 years and is the most representative of pervasive developmental disorders, with psychodevelopmental disorders characterized primarily by social interaction disorders, communication disorders and restricted, stereotypical, repetitive behaviors. In the present application, the childhood autism may include childhood autism, Asperge's syndrome, Rett's syndrome, childhood disorganized, atypical autism, and/or other unspecified pervasive developmental disorder. Childhood autism is a biologically-based mental developmental disorder caused by a variety of factors, and is a disease that occurs in individuals with genetic susceptibility under the influence of specific environmental factors. The infant patients have obstacles in social interaction, communication and the like, and have narrow interests, and tend to repeat stereotypical behavior patterns.
In the present application, the term "intellectual impairment (MR)" generally refers to a persistent impairment of cognitive activities and/or an impairment of overall psychological activities. The intellectual impairment may be due to organic damage to the brain caused by influencing factors (e.g. genetic variation, infection, poisoning, head injury, craniocerebral deformity or endocrine abnormalities) or due to incomplete brain development.
Detailed Description
High-intelligence horizontal mouse model and construction method and application thereof
In one aspect, the present application provides a method of constructing a high intellectual level mouse model, the method comprising replacing a chromosome in a mouse with all or part of a chromosome of wild type mouse origin, thereby making the mouse a high intellectual level mouse.
In the present application, the wild-type mouse may be derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
In the present application, the specific code numbers and strain designations of the wild-type mouse origin can be as shown in table 1.
For example, C57BL/6J represents a subline of C57BL of the Jackson laboratory Scale, USA, which may be abbreviated as B6/J.
In the present application, the mouse may be derived from strain C57 BL/6J.
In the present application, the replacement may comprise crossing with the wild type mouse.
In the present application, the replacement may include the steps of:
a) hybridizing a mouse with the wild-type mouse to obtain F1, backcrossing F1 with the mouse to obtain F2, and screening the mouse with at least one chromosome in F2 in a heterozygous state;
b) carrying out continuous backcross on the mouse with the chromosome in the heterozygous state obtained by screening in the step a) and the mouse, and screening to obtain a mouse population with the chromosome in the heterozygous state and the other chromosomes in the genotype of the homozygous mouse;
c) taking the mouse population obtained by screening in the step b) as an inbred line, and obtaining the mouse model through inbreeding.
The method of substitution may be known to those skilled in the art, and for example, a mouse having a substitution of
For example, the replacement may include the steps of:
(1) crossing ZZ2 mouse with mouse C57BL/6J to obtain F1 generation, backcrossing F1 generation mouse with C57BL/6J to obtain F2 generation, and screening out mouse with heterozygous state of whole No. 1 chromosome DNA sequence through genotyping;
(2) backcrossing the F2 generation mouse with C57BL/6J, backcrossing in the same way, and screening 10 generations to form a mouse population with a No. 1 chromosome DNA sequence in a heterozygous state and all the rest chromosomes being homozygous C57BL/6J genotypes;
(3) identifying the gene DNA of the backcrossed mice;
(4) after 10 generations of backcross, the backcross mouse with No. 1 chromosome ZZ2 and B6 in heterozygous state is bred into a homologous gene carrying No. 1 ZZ2 mouse, and the homologous gene is introduced into an inbred line, and through brother selfing and gene identification and screening, the mouse with high intelligence level (such as D000765 mouse) can be obtained.
For another example, the replacing may include the steps of:
(1) hybridizing a BLD mouse with a mouse C57BL/6J to obtain an F1 generation, backcrossing the F1 generation mouse with the C57BL/6J to obtain an F2 generation, and screening out the mouse with the whole No. 1 chromosome DNA sequence in a heterozygous state through genotyping;
(2) backcrossing the F2 generation mouse with C57BL/6J, backcrossing in the same way, and screening 10 generations to form a mouse population with a No. 1 chromosome DNA sequence in a heterozygous state and all the rest chromosomes being homozygous C57BL/6J genotypes;
(3) identifying the gene DNA of the backcrossed mice;
(4) after 10 generations of backcross, the backcross mouse with the BLD and B6 heterozygous for
In the present application, the screening may comprise genotyping. The genotyping may be a process of examining the DNA sequence of an individual by using a biological assay. For example, the genetic analysis may compare a sequence of interest to a sequence of another individual or to a reference sequence to determine differences in the genetic makeup (genotype) of the individuals. The genotyping method may be a method conventional in the art, and may include, for example, identifying Single Nucleotide polymorphisms (Single Nucleotide polymorphisms), Restriction Fragment Length Polymorphisms (RFLP), Terminal Restriction Length polymorphisms (t-RFLP), Amplified Fragment Length polymorphisms (Amplified Fragment h polymorphisms, AFLP), and/or Multiplex Ligation Probe Amplification (MLPA). The technical means for gene analysis may be any means conventional in the art, and may include, for example, Polymerase Chain Reaction (PCR), DNA fragment analysis, oligonucleotide probes (ASO probes), gene sequencing, nucleic acid hybridization, and/or gene chip technology.
In the present application, the screening may include identifying specific SNP sites of the chromosomes of the wild-type mouse. For example, whether
In the present application, the chromosome may include
In the present application, the replacement may include a means of utilizing gene editing. For example,
In the present application, said high intellectual level may be determined by evaluating one or more conditions of the group: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
In the present application, the emotional state may include an evaluation factor selected from the group consisting of: whether anxiety, whether depression, and whether caution is warranted. The high intelligence level mice described herein may be prudent, may be anxious, and/or may be non-depressive.
In the present application, the emotional state may be evaluated by a sugar water preference test. If the emotional state of the mouse presents problems and/or disorders, the bias towards sugar water is reduced; conversely, high mental level mice should maintain a normal level of carbohydrate bias. The preference degree of the mouse for sugar water can be evaluated by calculating a sugar water preference index, and then the emotional state of the mouse is evaluated: sugar water preference index% = sugar water consumption/(sugar water consumption + pure water consumption) × 100%.
In the present application, the learning capacity, and/or the memory capacity may be evaluated by a water maze test. The water maze test (such as Morris water maze) can force the mouse to swim, so as to learn and search for a platform hidden in water, and further test the learning ability and/or the memory ability of the mouse to the spatial position sense and the direction sense. The water maze test may also include an acquisition training, a scout training, an alignment training, or an alignment scout training. If the shorter the time required for a mouse to find a platform from entering the water, the shorter the distance traveled during that period, the mental level of the mouse will be correspondingly about high. The water maze assessment test may be an important test for assessing the mental level of a mouse.
In the present application, the spatial exploratory power, and/or the memory power, may be evaluated by a spatial exploratory Y maze test. The spatially probed Y maze may consist of three identical arms, each with a food supply at the end. When a mouse searches for food in the maze, the mouse can remember the arms of the maze which the mouse has searched according to the visual marks around the maze so as to avoid repeatedly entering the same arm, thereby effectively obtaining the food. The spatially probed Y maze test can evaluate the mouse's ability to recognize and remember new environments. Meanwhile, if the phenotype of the mouse in the space exploration Y maze test is more cautious, the higher intelligence level of the mouse can be reflected.
In the present application, the memory capacity can be evaluated by the active avoidance Y maze test. The active avoidance Y maze test may consist of three arms, some of which are relatively safe (i.e., safe zones) and others of which may be energized to generate sufficient current to escape the mouse. During the experiment, the safety zone can be changed, the mouse is electrically shocked, the safety zone that the mouse escapes immediately is taken as a correct reaction, and the mouse can continuously react for a plurality of times in the training process to obtain the 'scholarly'. The lower the number of exercises required to "learn" the higher the level of intelligence of the mouse. The active avoidance Y maze test can embody the mouse memory capacity to fear.
In the present application, the motor ability may be evaluated by an open field test and/or a fatigue swing rod test.
In the present application, the open field test is a method for evaluating the autonomous behavior of the experimental animal in a strange environment, and exploring behavior and tensity. If a mouse does not actively go to a central area (i.e., the mouse does not tend to be explored at will and/or be overstrained or overstrained in an unfamiliar environment), the mouse tends to be cautious, or the level of mental capacity of the mouse can be considered high. Meanwhile, the open field test can evaluate the motor capacity of the mouse, namely, the normal activity capacity and reaction speed of the mouse can be evaluated.
In the present application, the fatigue suspension bar test can evaluate the motor ability of the mouse, and particularly can evaluate the action coordination and the anti-fatigue property of the mouse. The high mental level mice are reflected by the results of the fatigue swing rod test, and the motor ability of the high mental level mice is not significantly different from that of the control group mice (such as C57BL/6J mice).
In another aspect, the present application provides a high mental level mouse model constructed by the methods described herein.
In another aspect, the present application provides a high mental-level mouse model in which the mouse chromosomes comprise at least all or part of the chromosomes of wild-type mouse origin.
In the present application, the wild-type mouse may be derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
In the present application, the mice may be derived from strains of the following group: c57 BL/6J.
In the present application, the mouse may comprise all or part of
In the present application, said high intellectual level may be determined by evaluating one or more conditions of the group: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
In the present application, the emotional state may be evaluated by a sugar water preference test.
In the present application, the learning capacity, and/or the memory capacity may be evaluated by a water maze test.
In the present application, the spatial exploratory power, and/or the memory power, may be evaluated by a spatial exploratory Y maze test.
In the present application, the memory capacity can be evaluated by the active avoidance Y maze test.
In the present application, the motor ability may be evaluated by an open field test and/or a fatigue swing rod test.
In the present application, the mouse in the high intelligence level mouse model (i.e. the high intelligence level mouse) can have higher intelligence level than the control group mouse (e.g. C57BL/6J mouse). For example, wherein the high mental level mouse may be more cautious, anxious, not depressed, have better emotional ability; the learning ability, especially the learning ability aiming at the spatial position sense and the direction sense, can be better; the memory capacity can be better, and the memory capacity can comprise the memory capacity aiming at the spatial position sense and the direction sense and can also comprise the memory capacity about fear; and/or may have better space exploration capabilities. For another example, the motor capacity of the high intellectual level mouse may not be significantly different from that of a control group of mice (e.g., C57BL/6J mice). Therefore, the mouse in the high intelligence level mouse model can have higher intelligence level under the condition of having stable and good emotional state.
In another aspect, the present application provides the use of a high mental level mouse model described herein for the preparation of a mouse model with a deficit in mental level.
In the present application, the system may be used to identify substances capable of affecting mental levels.
In another aspect, the present application provides a system for identifying an agent capable of affecting intellectual levels, the system comprising a high intellectual level mouse model in which the chromosomes of a mouse comprise at least all or part of a wild type mouse derived chromosome.
In the present application, the wild-type mouse may be derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
In the present application, the mouse may be derived from a strain selected from the group consisting of: c57 BL/6J.
In the present application, the mouse in the high mental level mouse model may comprise all or part of
In this application, the system may include a discrimination module that discriminates the effect of the substance on the mental level of the high mental level mouse model.
In the present application, the identification module may comprise reagents and/or instruments for identifying the effect of the substance on the mental level of the high mental level mouse model.
In the present application, the identification module may comprise reagents and/or instruments necessary to carry out an assay selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test. For example, the identification module may comprise a reagent and/or an instrument selected from the group consisting of: a sight box, sucrose solution, water (including dyed water), a water maze (e.g., including an underwater concealed platform), a Y maze (e.g., trisected radial labyrinth boxes), a trisected arm reflex box (e.g., including a pointing device), and a fatigue rotarod.
In the present application, the identification module may determine the effect of the substance on the activity and/or expression of the mental-level associated protein.
For example, the identification module may comprise reagents and/or apparatus capable of determining the activity and/or expression of the mental level associated protein. In the present application, the identification module may comprise reagents and/or instruments for determining the effect of the substance on the activity and/or expression level of the mental-level associated protein.
In the present application, the identification module may comprise an agent capable of specifically recognizing a mental level associated protein (e.g. the agent may comprise an antibody and/or ligand which specifically binds to the mental level associated protein), and/or a substance capable of determining the activity of a mental level associated protein (e.g. the substance may be an enzyme whose activity is associated with the activity of the mental level associated protein, and/or a protein and/or transcription factor whose activity is associated with the pathway of reactions upstream and downstream of the mental level associated protein).
In the present application, the identification module may comprise reagents and/or apparatus for determining the effect of the substance on the activity and/or expression level of a nucleic acid encoding a mental-level associated protein.
For example, the identification module may comprise reagents and/or apparatus capable of determining the activity and/or expression of a nucleic acid encoding the mental level associated protein. In the present application, the identification module may comprise a primer capable of specifically amplifying a nucleic acid encoding a mental level associated protein, and/or a probe capable of specifically recognizing a nucleic acid encoding a mental level associated protein (e.g., the probe may be a DNA double-stranded probe, a DNA single-stranded probe, an aptamer, or RNAi).
In another aspect, the present application provides a method of assessing the effect of a substance on mental levels, which method may comprise administering a candidate substance to a high mental level mouse model in which the chromosomes of the mouse comprise at least all or part of a wild-type mouse-derived chromosome.
In the present application, the mouse in the high mental level mouse model may comprise all or part of
In the present application, the wild-type mouse may be derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
In the present application, the mouse may be derived from a strain selected from the group consisting of: c57 BL/6J.
In the present application, the administration may include injection, oral administration, and/or external application.
In the present application, the substance may be a candidate drug for the prophylaxis and/or treatment of a disease associated with mental levels. For example, the mental level related disorder may include: alzheimer's disease, chronic brain disease syndrome, attention deficit disorder, childhood autism, and intellectual impairment (MR).
In the present application, the method may comprise the steps of: assessing the effect of the substance on the mental level of the high mental level mouse model.
In the present application, the intellectual level may be determined by evaluating one or more conditions of the group: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability. For example, the mental level may be determined by conducting a test selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.
In the present application, the assessing may comprise assessing the effect of the substance on the mental level of the high mental level mouse model. For example, a substance may be considered to reduce and/or harm the mental level of a mouse in the high mental level mouse model if, after administration of the substance, the mental level of the mouse in the high mental level mouse model decreases significantly. For another example, a substance may be considered not to reduce and/or harm the mental level of a mouse in the high mental level mouse model if, after administration of the substance, the mental level of the mouse in the high mental level mouse model is not significantly altered and/or reduced. The substance may also be considered safe for mice in the intellectual level field. For another example, a substance may be considered to increase the mental level of a mouse in a high mental level mouse model if, after administration of the substance, the mental level of the mouse in the high mental level mouse model is significantly increased.
The systems and/or methods described herein may be used to screen and/or assess whether the substance negatively affects mental performance; may also be used for security checks.
In the present application, the assessing may comprise assessing the effect of the substance on the activity and/or expression of the mental-level associated protein.
For example, the effect of an agent capable of specifically recognizing a mental level associated protein (e.g., the agent may comprise an antibody and/or ligand that specifically binds to the mental level associated protein), and/or a substance capable of determining the activity of a mental level associated protein (e.g., the substance may be an enzyme whose activity is associated with the activity of the mental level associated protein, and/or a protein and/or transcription factor whose activity is associated with the upstream and downstream pathways of the mental level associated protein) may be used to assess the activity and/or expression of the mental level associated protein.
In the present application, the assessing may comprise assessing the effect of the substance on the activity and/or expression of a nucleic acid encoding the mental level associated protein.
For example, the effect of the agent on the activity and/or expression of a nucleic acid encoding a mental level associated protein may be assessed by using an agent and/or apparatus capable of determining the activity and/or expression of a nucleic acid encoding the mental level associated protein. For example, the agent may include a primer capable of specifically amplifying a nucleic acid encoding a mental level associated protein, and/or a probe capable of specifically recognizing a nucleic acid encoding a mental level associated protein (e.g., the probe may be a DNA double-stranded probe, a DNA single-stranded probe, an aptamer, or RNAi).
The present application also relates to the following embodiments:
1. a method of constructing a high mental level mouse model, the method comprising replacing a chromosome in a mouse with all or part of a chromosome derived from a wild type mouse, thereby making the mouse a high mental level mouse.
2. The method of
3. The method of any one of embodiments 1-2, wherein the mouse is derived from a strain selected from the group consisting of: c57 BL/6J.
4. The method of any one of embodiments 1-3, wherein said replacing comprises crossing with said wild-type mouse.
5. The method according to any one of embodiments 1-4, wherein said replacing comprises using means of gene editing.
6. The method according to any of embodiments 1-5, wherein said replacing comprises the steps of:
a) hybridizing a mouse with the wild-type mouse to obtain F1, backcrossing F1 with the mouse to obtain F2, and screening the mouse with at least one chromosome in F2 in a heterozygous state;
b) carrying out continuous backcross on the mouse with the chromosome in the heterozygous state obtained by screening in the step a) and the mouse, and screening to obtain a mouse population with the chromosome in the heterozygous state and the other chromosomes in the genotype of the homozygous mouse;
c) taking the mouse group obtained by screening in the step b) as an inbred line, and obtaining the mouse model through inbreeding.
7. The method of embodiment 6, wherein said screening comprises genotyping.
8. The method of any one of embodiments 6-7, wherein the screening comprises identifying specific SNP sites of the chromosome of the wild-type mouse.
9. The method of any one of embodiments 1-8, wherein the chromosome comprises
10. The method according to any one of embodiments 1-9, wherein said high intellectual level is determined by evaluating one or more conditions of the group consisting of: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
11. The method according to
12. The method according to any one of embodiments 10-11, wherein the learning capacity, and/or the memory capacity is evaluated by a water maze test.
13. The method according to any one of embodiments 10-12, wherein said spatial exploratory capacity, and/or said memory capacity, is assessed by a spatially exploratory Y maze test.
14. The method according to any one of embodiments 10-13, wherein the memory capacity is assessed by the active avoidance Y maze test.
15. The method of any one of embodiments 10-14, wherein the athletic ability is evaluated by an open field test and/or a fatigue hover rod test.
16. A high mental level mouse model constructed by the method of any one of embodiments 1-15.
17. A high mental level mouse model in which the mouse chromosome comprises at least all or part of a wild type mouse derived chromosome.
18. The high intellectual level mouse model of embodiment 17, wherein the wild type mouse is derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
19. The high intellectual level mouse model according to any one of embodiments 17 to 18, wherein the mouse is derived from a strain selected from the group consisting of: c57 BL/6J.
20. A high intellectual level mouse model according to any one of embodiments 17 to 19 wherein the mouse comprises all or part of
21. The high intellectual level mouse model according to any one of embodiments 17-20, wherein the high intellectual level is determined by evaluating one or more conditions of the group consisting of: motor ability, emotional state, learning ability, memory ability, and spatial exploration ability.
22. The method of embodiment 21, wherein the emotional state is assessed by a sugar water preference test.
23. The high intelligence level mouse model of any one of embodiments 21-22, wherein the learning ability, and/or the memory ability is evaluated by a water maze test.
24. The high intelligence level mouse model of any one of embodiments 21-23, wherein the learning ability, and/or the memory ability is tested by a water maze assessment.
25. The high intellectual level mouse model according to any one of embodiments 21-24, wherein the spatial exploratory ability, and/or the memory ability, is evaluated by a spatial exploratory Y maze test.
26. The high intellectual level mouse model according to any one of embodiments 21-25, wherein the memory capacity is assessed by an active avoidance Y maze test.
27. The high mental level mouse model method according to any one of embodiments 21-26, wherein the motor ability is evaluated by an open field test and/or a fatigue spinning rod test.
28. Use of a high mental level mouse model according to any one of embodiments 17-27 for the preparation of a mouse model with a deficit in mental level.
29. Use of a high intelligent level mouse model according to any one of embodiments 17-27 for the preparation of a system for identifying a substance capable of affecting an intelligent level.
30. A system for identifying an agent capable of affecting intellectual levels, the system comprising a high intellectual level mouse model in which the chromosomes of a mouse comprise at least all or part of a wild type mouse derived chromosome.
31. The system of
32. The system of any one of embodiments 30-31, wherein the mouse is derived from a strain selected from the group consisting of: c57 BL/6J.
33. The system according to any one of embodiments 30-32, wherein the mouse in the high intellectual level mouse model comprises all or part of
34. A system according to any one of embodiments 30-33, wherein said system comprises an identification module that identifies an effect of said agent on mental levels of said high mental level mouse model.
35. The system of embodiment 34, wherein said identification module comprises reagents and/or instruments that identify the effect of said substance on the mental level of said high mental level mouse model.
36. The system of any one of embodiments 34-35, wherein the identification module comprises reagents and/or instrumentation required to perform an assay selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.
37. The system according to any one of embodiments 34-36, wherein the identification module determines the effect of the agent on the activity and/or expression of an intellectual-level associated protein.
38. The system according to any one of embodiments 34-37, wherein said identification module comprises reagents and/or instruments for determining the effect of said substance on the activity and/or expression level of an intellectual-level associated protein.
39. A system according to any one of embodiments 34-38, wherein the identification module comprises an agent capable of specifically recognizing a mental level associated protein, and/or a substance capable of determining the activity of a mental level associated protein.
40. The system according to any one of embodiments 34-39, wherein said identification module comprises reagents and/or instruments for determining the effect of said agent on the activity and/or expression level of a nucleic acid encoding a mental-level associated protein.
41. The system according to any one of embodiments 34-40, wherein the identification module comprises a primer capable of specifically amplifying a nucleic acid encoding a mental level associated protein, and/or a probe capable of specifically recognizing a nucleic acid encoding a mental level associated protein.
42. A method of assessing the effect of a substance on mental capacity, the method comprising administering a candidate substance to a high mental capacity mouse model in which the chromosomes of the mouse comprise at least all or part of a wild type mouse derived chromosome.
43. The method of embodiment 42, wherein the wild-type mouse is derived from a strain selected from the group consisting of: BLD, CM, DX, FX, HZ, JD, KM, PD, QP, SMX, SY, TW, TZ, YP, YX, ZC, ZZ1, and
44. The method of any one of embodiments 42-43, wherein the mouse is derived from a strain selected from the group consisting of: c57 BL/6J.
45. The method according to any one of embodiments 42-44, wherein the mouse in the high intellectual level mouse model comprises all or part of
46. The method according to any one of embodiments 42-45, wherein said administering comprises injection, oral administration, and/or topical administration.
47. The method of any one of embodiments 42-46, comprising the steps of: assessing the effect of the substance on the mental level of the high mental level mouse model.
48. The method of embodiment 47, wherein said assessing comprises assessing the effect of said substance on the mental level of said high mental level mouse model.
49. The method of any one of embodiments 47-48, wherein said assessing comprises conducting an assay selected from the group consisting of: the method comprises an open field test, a sweet water preference test, a water maze test, a space exploration Y maze test, an active avoidance Y maze test and a fatigue suspension rod test.
50. The method according to any one of embodiments 47-49, wherein said assessing comprises assessing the effect of said agent on the activity and/or expression of an intellectual-level associated protein.
51. The method according to any one of embodiments 47-50, wherein said assessing comprises assessing the effect of said agent on the activity and/or expression of a nucleic acid encoding a mental-level associated protein.
Without intending to be bound by any theory, the following examples are merely intended to illustrate the fusion proteins, preparation methods, uses, etc. of the present application, and are not intended to limit the scope of the invention of the present application.
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