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A method for establishing an animal model of non-human mammalian anxiety or related diseases and its application

A technology for non-human mammals and animal models, which is applied in the field of establishing animal models of non-human mammalian anxiety disorders or related diseases, and can solve problems such as the increase in the proportion of physical stress.

Active Publication Date: 2020-01-17
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this type of model contains components of physical stress such as electric shock, making the increase in the proportion of physical stress in psychological stress is its main defect

Method used

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  • A method for establishing an animal model of non-human mammalian anxiety or related diseases and its application
  • A method for establishing an animal model of non-human mammalian anxiety or related diseases and its application
  • A method for establishing an animal model of non-human mammalian anxiety or related diseases and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0141] Example 1 Establishment of a mouse animal model in which the Glce gene is knocked out in neuron-specific conditions.

[0142] First construct the Glce mutant gene sequence ( figure 1 ). Such as figure 1 As shown, the Glce gene knockout targeting vector sequence was designed, the Loxp / Loxp allele was inserted on both sides of exon 3 of the Glce gene, and the neo gene was inserted at the 3' end, with a 5' end arm of 3125 bp and a 3' end arm of 3718 bp.

[0143] figure 2 Plasmid map of targeting vector for Glce gene knockout. 1. Obtain the homologous fragment of the target gene (Glce), and clone the DNA fragment into the plasmid vector; 2. Excise most of the homologous DNA sequences of the target gene from the recombinant plasmid, leaving only part of the sequence on both sides of the online plasmid vector. 3. Cloning the neo gene into a linearized plasmid with the homologous sequence of the target gene so that it is located in the middle of the homologous sequence of...

Embodiment 2

[0178] Behavioral Analysis of Embodiment 2 Glce Mutant Homozygous Mice

[0179] Glce gene mutant homozygous mice carrying Cre recombinase (hereinafter referred to as Glce mutant homozygous mice) were raised in a clean feeding environment. A series of animal behavior experiments including autonomous activity, open field test, elevated plus maze and other experiments were used to evaluate the anxiety-like symptoms of the mouse animal model in the adolescent period of mice (>3 months old).

[0180] 2.1 Autonomous activities

[0181] The mice were placed in a dark experiment box (110mm X 110mm X 330mm), and the spontaneous activity of the mice was tested for 5 minutes. The activity of mice was captured by infrared camera. Use the video tracking software and analysis software of Shanghai Jiliang Software Technology Co., Ltd. to analyze the mouse activity trajectory and activity time.

[0182] The study found that compared with the young C57BL / 6 normal mice, there was no signific...

Embodiment 3

[0191] Depression Behavioral Analysis of Example 3 Glce Mutant Homozygous Mice

[0192] 3.1 Autonomous activities

[0193] The Glce gene mutation homozygous mice carrying Cre recombinase were placed in a dark experimental box (110mm X 110mm X 330mm), and the spontaneous activity of the mice was tested for 5 minutes. The activity of mice was captured by infrared camera. Use the video tracking software and analysis software of Shanghai Jiliang Software Technology Co., Ltd. to analyze the mouse activity trajectory and activity time.

[0194] Studies have shown that compared with young C57BL / 6 normal mice, there is no significant difference in the total distance of spontaneous activities, activity time and activity speed of adolescent Glce mutant homozygous mice ( Figure 11 ).

[0195] The results showed that the innate behavior of adolescent Glce mutant homozygous mice was not significantly different from that of C57BL / 6 normal mice.

[0196] 3.2 Open field experiment

[01...

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Abstract

The invention provides an establishment method and application of an anxiety disorder or related disease animal model of non-human mammals and in particular provides a preparation method of the anxiety disorder or related disease animal model of the non-human mammals. The preparation method comprises the following steps: (a) providing cells of the non-human mammals and inactivating Glce genes in the cells to obtain non-human mammal cells with inactivated Glce genes; (b) preparing the anxiety disorder or related disease animal model with the inactivated Glce genes through utilizing the cells with the inactivated Glce genes, which are prepared in the step (a). The animal model provided by the invention is an effective anxiety disorder or related disease animal model and can be widely used for researching diseases including generalized anxiety, acute anxiety attack, phobic disorder, post-traumatic stress disorder, acute stress disorder, obsessive-compulsive disorder and the like, and can be used for screening and testing specific drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for establishing an animal model of non-human mammalian anxiety or related diseases and its use. Background technique [0002] Anxiety disorder, also known as anxiety neurosis, is the most common type of neurosis. It is characterized by anxiety and its core symptom is worry. Mainly include generalized anxiety, acute anxiety attacks, phobias, post-traumatic stress disorder, acute stress disorder, and obsessive-compulsive disorder. The main manifestations are nervousness and worry without clear objective objects, restlessness, and autonomic symptoms (palpitations, hand tremors, sweating, frequent urination, etc.). [0003] World Health Organization (WHO) statistics show that from 1990 to 2003, the number of people suffering from depression or anxiety increased from 416 million to 615 million, almost reaching 10% of the global population. Also, mental disorders account for 30...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/00C12N5/10A01K67/027
CPCA01K67/0278A01K2207/15A01K2217/075A01K2227/105A01K2227/106A01K2227/107A01K2267/0356A61K49/0008C12N5/0606C12N9/90C12N2510/00C12Y503/01017
Inventor 丁侃李艳玲
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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