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Establishment method of animal model for phobia or phobia-relevant diseases of non-human mammals and application of the animal model

A technology of non-human mammals and animal models, which is applied in the field of establishment of animal models of non-human mammal phobias or related diseases, and can solve problems such as difficult practical operations, inability to fully simulate primary phobias, and complicated surgery

Active Publication Date: 2017-01-04
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

[0005] Most of the existing mouse phobia models currently have to be treated with exogenous drugs or other physical and chemical methods, or even surgical means, which cannot completely simulate human primary phobias, and there are such problems as surgery. Complicated, difficult actual operation, long modeling time, unstable effect, large individual differences, etc.

Method used

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  • Establishment method of animal model for phobia or phobia-relevant diseases of non-human mammals and application of the animal model
  • Establishment method of animal model for phobia or phobia-relevant diseases of non-human mammals and application of the animal model
  • Establishment method of animal model for phobia or phobia-relevant diseases of non-human mammals and application of the animal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Embodiment 1 obtains the homozygous mouse of Glce gene mutation carrying Cre recombinase

[0133] First construct the Glce mutant gene sequence ( figure 1 ). Such as figure 1 The Glce gene deletion targeting vector sequence is designed as shown, the Loxp / Loxp allele is inserted into both sides of exon 3 of the Glce gene, and the neo gene is inserted at the 3' end, the 5' end arm is 3125bp, and the 3' end arm is 3718bp. 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 the residual target gene; 4. Linearizing ...

Embodiment 2

[0167] Fear Behavioral Analysis of Example 2 Glce Mutant Homozygous Mice

[0168] 2.1 Autonomous activities

[0169] 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.

[0170] 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 8 ).

[0171] The results showed that the innate behavior of adolescent Glce mutant homozygous mice was not significantly different from that of C57BL / 6 normal mice.

[0172] 2.2 Open field experiment

[0173] The...

Embodiment 3

[0180] Example 3 Anxiety Behavioral Analysis of Glce Mutant Homozygous Mice

[0181] 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).

[0182] 3.1 Autonomous activities

[0183] 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.

[0184] The study found that compared with the young C57BL / 6 normal mice, there was no sig...

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Abstract

The invention provides an establishment method of an animal model for phobia or phobia-relevant diseases of non-human mammals and application of the animal model and particularly provides a preparation method of the animal model. The preparation method comprises the following steps: (a) providing non-human mammal cells, and inactivating Glce genes in the cells, so as to obtain the non-human mammal cells with the inactivated Glce genes; and (b) preparing the animal model with the inactivated Glce genes for the phobia or the phobia-relevant diseases by virtue of the cells with the inactivated Glce genes prepared in the step (a). The animal model is an effective animal model for the phobia or the phobia-relevant diseases, can be used for researching diseases of the phobia, social phobia, topophobia and the like and further can be applied to the screening and tests of 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 mammal phobia or related diseases and its use. Background technique [0002] Phobia is a kind of neurosis with fear symptom as the main clinical manifestation. Patients have strong and unnecessary fear of certain specific objects or situations, accompanied by obvious anxiety and autonomic symptoms, and actively adopt avoidance methods to relieve this anxiety. The core symptom of phobias is fear and tension, which can cause severe anxiety and even panic. [0003] At present, the pathogenic mechanism of phobia has not been studied clearly, and there is a lack of safe and effective treatment methods. Therefore, an in-depth study of its pathogenesis is an important basis for the treatment of phobia. Mouse disease models play a very important role in the study of the pathogenic mechanism of human diseases and drug screening. Mouse ...

Claims

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Application Information

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IPC IPC(8): C12N5/10A01K67/027A01K49/00
CPCA01K67/0271A01K67/0276A01K2207/12A01K2207/15A01K2217/075A01K2227/105A01K2227/106A01K2227/107A01K2267/03A61K49/00A61K49/0008C12N9/90C12N15/8509C12N2800/107C12Y501/03A01K67/027C12N5/10
Inventor 丁侃李艳玲
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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