CN114304064A - Method for establishing animal model with head deformity accompanied with nerve-dyskinesia - Google Patents
Method for establishing animal model with head deformity accompanied with nerve-dyskinesia Download PDFInfo
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- CN114304064A CN114304064A CN202111536611.4A CN202111536611A CN114304064A CN 114304064 A CN114304064 A CN 114304064A CN 202111536611 A CN202111536611 A CN 202111536611A CN 114304064 A CN114304064 A CN 114304064A
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Abstract
A method for establishing a head deformity with nerve-dyskinesia model is disclosed, which belongs to the application in the field of medicine, in particular to a head deformity with nerve-dyskinesia model for screening nerve-dyskinesia medicines, a construction method and application. A spontaneous head and body abnormality accompanying nerve-dyskinesia mouse model is established by adopting a gene knockout technology and screening 3-week-old head and body proportion abnormality and 8-week-old ethological screening respectively, and the head and body proportion of the animal offspring is abnormal, a large amount of liquid is accumulated in the brain, the animal cannot accurately move straight and turn round in situ, and the brain section is abnormal. Has obvious nerve-movement disorder. The head deformity with nerve-dyskinesia injury model established by the invention is generated spontaneously, can be used for animal models of young mouse born nerve system injury and related head deformity with nerve-dyskinesia, can be used for testing various medicaments and combined medicaments, and can effectively screen the medicaments.
Description
Technical Field
The invention relates to the field of establishment of an animal model with head deformity and nerve-dyskinesia, in particular to an establishment method of a standardized animal model with head deformity and nerve-dyskinesia.
Background
Birth defects refer to the presence of dysplasia and deformities of certain parts of the body of the mother's uterus that occur in the mother prior to birth. The head deformity and the cerebral edema are located in the first 5 places of the birth defects in China. Each year, 80-120 million birth defects are born, accounting for 4-6% of all the population born.
However, the unclear mechanism of head deformity and dyskinesia of offspring and the absence of animal models are the difficulties of the current drug regulation.
The scavenger receptor type B I (SR-BI), a high density lipoprotein receptor, selectively takes up high density lipoproteins in the liver and carries excess cholesterol in the body to the liver for excretion. Studies have shown that SR-BI is expressed in nerve cells and has beta-amyloid binding activity, possibly affecting nervous system diseases.
At present, there is a significant animal model deletion of head deformity with neuro-dyskinesia.
Disclosure of Invention
The invention mainly aims to provide a method for establishing a standardized animal model with head deformity and nerve-dyskinesia, which can effectively solve the problems in the background art.
The invention obtains homozygote or heterozygote parents by carrying out scavenger receptor B type I gene knockout technology; selecting homozygote or heterozygote parents for propagation; and (5) screening the filial generation for abnormal head-body ratio, wherein the head-body ratio is higher than 0.2.
The invention obtains homozygote or heterozygote parents by carrying out scavenger receptor B type I gene knockout technology, selects homozygote or heterozygote parents to breed, carries out ethological abnormality screening on the offspring on the basis of head-body proportion abnormality screening, and has obvious behaviors of incapability of going straight and circling.
The invention obtains homozygote or heterozygote parents by carrying out scavenger receptor B type I gene knockout technology, selects homozygote or heterozygote parents to breed, and a progeny section observes that the number of hippocampal dentate gyrus neuron cells is obviously reduced compared with the wild type group, a large number of vacuoles, cytoplasm shrinkage, cell nucleus shrinkage, thalamus blood vessel number is obviously increased, neuron cell cytoplasm shrinkage, cell nucleus shrinkage, cerebellum purkinje cell nucleus shrinkage, granular cell gap enlargement and molecular layer blood vessel number increase, and the obvious hydrocephalus of the brain can be seen by MRI scanning.
The invention obtains homozygote or heterozygote parents by carrying out scavenger receptor B type I gene knockout technology, selects homozygote or heterozygote parents to breed, and obtains obvious hydrocephalus of brain by MRI scanning of filial generation.
The head deformity with nerve-dyskinesia injury model established by the invention is generated spontaneously, has obvious nerve-dyskinesia, and can be used for animal models of the nerve system injury of young mice born and the related head deformity with nerve-dyskinesia.
The animal model with the characteristics of head deformity with nerve-dyskinesia can be used for testing various medicaments and combined medicaments and effectively screening the medicaments.
Drawings
FIG. 1: example 1 of the invention compares the result chart with the abnormal head-to-body ratio
FIG. 2: example 2 behavioral anomaly result graph of the invention
FIG. 3: example 3 Hippocampus dentate gyrus staining result of brain section of the invention
FIG. 4: example 3 thalamic staining result chart of brain section of the present invention
FIG. 5: example 3 cerebellar Purkinje staining result chart of brain section
FIG. 6: example 4 of the present invention is a brain MRI scan result chart.
Detailed Description
Example 1 screening for abnormal head-to-body ratio in model animals
Selecting a C57 mouse, and carrying out scavenger receptor B type I gene knockout technology under the culture conditions: the laboratory temperature is 22-25 ℃, and the relative humidity is as follows: 40-70%, and light circulation: 12 hours of illumination (8:00-20:00), 12 hours of darkness (20: 00-8: 00), animal feeding: 2 mice/cage, central air-conditioning and air-filtering mechanical equipment, sterilizing animal cages, drinking water, padding and the like by high-pressure steam, feeding special SPF-level granulated feed, freely drinking water, selecting male homozygote to be matched with female homozygote, or male heterozygote to be matched with female homozygote, breeding, screening the filial generation with abnormal head-body proportion in 3-week-old filial generation, and screening animals with obvious short body state and abnormal head-body proportion which is more than or equal to 0.2 compared with wild congeneric mice (figure 1).
Example 2 screening for head-to-body scale abnormalities in model animals
Selecting a C57 mouse, and carrying out scavenger receptor B type I gene knockout technology under the culture conditions: the laboratory temperature is 22-25 ℃, and the relative humidity is as follows: 40-70%, and light circulation: 12 hours of illumination (8:00-20:00), 12 hours of darkness (20: 00-8: 00), animal feeding: 2 mice/cage, central air-conditioning and air-filtering mechanical equipment, sterilizing animal cages, drinking water, padding and the like by high-pressure steam, feeding special SPF-level granulated feed, freely drinking water, selecting male homozygote to be matched with female homozygote, or male heterozygote to be matched with female homozygote, breeding, screening the filial generation with abnormal head-body proportion at the age of 3 weeks, screening the animals with obvious short body state and abnormal head-body proportion compared with wild same-age mice, screening the animals with abnormal behavior at the age of 8 weeks, screening the animals with obvious abnormal behavior, which can not go straight and turn round in place (figure 2).
Example 3 screening for abnormal head-to-body ratio in model animals
Selecting a C57 mouse, and carrying out scavenger receptor B type I gene knockout technology under the culture conditions: the laboratory temperature is 22-25 ℃, and the relative humidity is as follows: 40-70%, and light circulation: 12 hours of illumination (8:00-20:00), 12 hours of darkness (20: 00-8: 00), animal feeding: 2 mice/cage, central air-conditioning and air-filtering mechanical equipment, sterilizing animal cages, drinking water, padding and the like by high-pressure steam, feeding special SPF-level granulated feed, freely drinking water, selecting male homozygote to be matched with female homozygote, or male homozygote to be matched with female heterozygote, or male heterozygote to be matched with female homozygote, breeding, screening off filial generation with abnormal head-body proportion at 3 weeks of age of the filial generation, screening animals with obvious short body state and abnormal head-body proportion compared with wild congeneric mice, and screening off abnormal behavior at 8 weeks of the filial generation. The screened animals are stained with brain slices, and the HE staining result shows that the number of hippocampal dentate gyrus neuron cells is obviously reduced compared with that of wild type groups, a large number of vacuoles are generated near the neurons, cytoplasm is shrunk, cell nucleus is shrunk (figure 3), the number of thalamus blood vessels is obviously increased, the number of neuron cells is shrunk, cell nucleus is shrunk (figure 4), cell nucleus of cerebellum Purkinje is shrunk, the particle intercellular space is enlarged, the number of molecular layer blood vessels is increased (figure 5), and the obvious hydrocephalus of the brain can be seen by MRI scanning.
Example 4 model animals are screened for abnormal head to body ratios.
Selecting a C57 mouse, and carrying out scavenger receptor B type I gene knockout technology under the culture conditions: the laboratory temperature is 22-25 ℃, and the relative humidity is as follows: 40-70%, and light circulation: 12 hours of illumination (8:00-20:00), 12 hours of darkness (20: 00-8: 00), animal feeding: 2 mice/cage, central air-conditioning and air-filtering mechanical equipment, sterilizing animal cages, drinking water, padding and the like by high-pressure steam, feeding special SPF-level granulated feed, freely drinking water, selecting male homozygote to be matched with female homozygote, or male homozygote to be matched with female heterozygote, or male heterozygote to be matched with female homozygote, breeding, screening the filial generation with abnormal head-body proportion at the age of 3 weeks of the filial generation, screening the animals with obvious short body state and abnormal head-body proportion compared with wild congeneric mice, screening the animals with obvious abnormal behavior at the age of 8 weeks of the filial generation, screening the animals which have obvious abnormal behavior and can not go straight, and circling the animals in situ. The screened animals were stained with brain sections, HE staining showed abnormal section display, and MRI scan showed significant hydrocephalus in the brain (fig. 6).
The standardized animal model building method of head deformity with nerve-dyskinesia is realized by a scavenger receptor B type I gene knockout technology; selecting homozygote or heterozygote parents for propagation; screening head-body proportion abnormality and screening behavioral abnormality of the offspring; the number of hippocampal dentate gyrus neuron cells is obviously reduced compared with that of wild type groups through slice observation, a large number of vacuoles appear near the neurons, cytoplasm is shrunk, cell nucleus is shrunk, the number of thalamus blood vessels is obviously increased, the cytoplasm of the neuron cells is shrunk, the cell nucleus of cerebellum purkinje is shrunk, the particle intercellular space is enlarged, the number of molecular layer blood vessels is increased, and obvious hydrocephalus of the brain can be seen through MRI scanning. The nerve-dyskinesia injury model established by the invention is generated spontaneously, has obvious nerve-dyskinesia and can be used for the animal model of the nerve system injury of the newborn mice and the related nerve-dyskinesia. The obtained animal model with the characteristics of head deformity with nerve-dyskinesia can be used for testing various drugs and combined drugs and effectively screening the drugs.
The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and the embodiments and descriptions are only illustrative of the principles of the invention, and all conventional changes, substitutions, or combinations of technical details disclosed herein are within the scope of the claimed invention. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. A method for establishing an animal model with head deformity accompanied with nerve-dyskinesia is characterized in that:
adopting a C57 mouse as an animal body; performing gene knockout technology; selecting homozygote or heterozygote parents for breeding, screening the offspring for behavioral abnormality with the head-body proportion abnormality of more than or equal to 0.2, screening the offspring for behavioral abnormality, taking offspring of 8-week-old animal with behavioral abnormality, slicing and observing hippocampal dentate gyrus, thalamus dentate gyrus and cerebellar Purkinje cells, taking offspring of 8-week-old animal with behavioral abnormality, and observing obvious brain abnormality by MRI scanning.
2. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: the gene knockout technology is the knockout of class B I type genes of scavenger receptors.
3. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: selecting homozygote or heterozygote parents for propagation.
4. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: screening for abnormal proportion of head and body in 3 weeks of age of offspring.
5. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: the offspring were screened for behavioral abnormalities at 8 weeks of age.
6. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: offspring brain sections were observed for brain abnormalities.
7. The method for establishing an animal model with head deformity and neuro-dyskinesia according to claim 1, wherein: significant brain abnormalities were visible on MRI scans.
8. An animal model with head malformation accompanied with nerve-dyskinesia for screening nerve-dyskinesia drugs, which is obtained by the method for constructing an animal model with nerve-dyskinesia for screening nerve-dyskinesia drugs according to any one of claims 1 to 8.
9. The application of the nerve-dyskinesia animal model is characterized in that the nerve-dyskinesia animal model for screening the medicines for treating head deformity accompanied with nerve-dyskinesia, which is obtained by the construction method of any one of claims 1 to 8, is used for screening the medicines for regulating nerve-dyskinesia.
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2021
- 2021-12-16 CN CN202111536611.4A patent/CN114304064A/en active Pending
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| US20020108131A1 (en) * | 1999-06-28 | 2002-08-08 | Monty Krieger | SR-BI and apo E knockout animals and use thereof as models for atherosclerosis and heart attack |
| US20050223420A1 (en) * | 2004-04-05 | 2005-10-06 | Massachusetts Institute Of Technology Commonwealth Of Massachusetts | Inducible heart attack animal model |
| CN107849540A (en) * | 2015-01-28 | 2018-03-27 | 淡马锡生命科学研究院有限公司 | Enterovirus 71 animal model |
| US20190289835A1 (en) * | 2016-09-09 | 2019-09-26 | Beth Israel Deaconess Medical Center, Inc | Mouse Models Having a Knockin Scavenger Receptor Class B Type I |
| CN108324945A (en) * | 2017-01-19 | 2018-07-27 | 首都医科大学附属北京妇产医院 | For inhibiting Nano medication particle to penetrate the inhibitor of placental barrier |
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