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Mouse model of spermatogenesis disorder with ddx5 gene deletion and its construction method

A mouse model, construction method technology, applied in the biological field to avoid damage and interference

Active Publication Date: 2022-03-22
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, no studies have reported the relationship between the Ddx5 gene and spermatogenesis or reproductive dysfunction

Method used

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  • Mouse model of spermatogenesis disorder with ddx5 gene deletion and its construction method
  • Mouse model of spermatogenesis disorder with ddx5 gene deletion and its construction method
  • Mouse model of spermatogenesis disorder with ddx5 gene deletion and its construction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Obtaining germline-specific knockout Ddx5 mice

[0040] 1. Experimental animals

[0041] Vasa-Cre transgenic mice expressing exogenous Cre driven by germ cell-specific genes: the strain is C57BL / 6, donated by Wu Ji of Shanghai Jiao Tong University and Li Wei of Institute of Zoology, Chinese Academy of Sciences, 4 females. floxylated Ddx5 transgenic mice: Ddx5 flox / flox , the strain is C57BL / 6, donated by Fan Ye, Xinqiao Hospital, Third Military Medical University, with 2 males. The mice were fed according to the SPF animal feeding standards.

[0042] 2. Obtain germline-specific knockout Ddx5 mice

[0043] Schematic diagram of reproductive strategy for germline-specific knockout Ddx5 mice figure 1 As shown, the specific steps are as follows:

[0044] (1) Vasa-Cre, a transgenic mouse expressing exogenous Cre driven by a germ cell-specific gene, and a floxed Ddx5 transgenic mouse (Ddx5 flox / flox ) mating to obtain the F1 generation;

[0045] (2) Get the to...

Embodiment 2

[0060] Example 2 Reproductive organ pathological morphology detection

[0061] Observe the morphology of the testes of the reproductive organs of male mice with germline-specific Ddx5 knockout in the F2 generation. The result is as image 3 As shown, the results show that the germline-specific knockout of Ddx5 has no significant difference in body shape and appearance between mice and wild-type male mice ( image 3 A), but compared with the wild type, the testis size of the germline-specific knockout Ddx5 mice (Figure 3B) is significantly reduced. Take 5 wild-type and germline-specific knockout Ddx5 male mice, and weigh the testes , the testis weight of knockout mice was significantly reduced ( image 3 C), where P is less than 0.001, with a very significant difference.

[0062] Histopathological sections (testes and epididymis) and H&E staining (hematoxylin-eosin staining) of reproductive organs of male mice with germline-specific Ddx5 knockout, the results are as follows ...

Embodiment 3

[0063] Example 3 Fertility Detection

[0064] Littermate wild-type male mice and germline-specific Ddx5-knockout male mice were mated with normal wild-type female mice after sexual maturity; the plugged female mice were placed alone, and the birth rate was observed. The results are shown in Table 5:

[0065]

[0066]

[0067] It can be seen from the above table that wild-type male mice were mated with 17 female mice, and a total of 138 pups were obtained, with an average of 8.12 pups per litter, while germline-specific knockout Ddx5 male mice were mated with 15 female mice, and no pups were obtained. offspring. The result statistics P is less than 0.001, which has a very significant difference. It can be seen that the germline-specific Ddx5 knockout male mice constructed in the present invention are indeed infertile, and can be used as a mouse model of spermatogenesis disorder for research.

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Abstract

The present invention provides a spermatogenesis disorder mouse model with Ddx5 gene deletion and its construction method. The inventors found in research that there is a relationship between Ddx5 gene and spermatogenesis disorder or reproductive dysfunction, and the male mice with Ddx5 gene deletion The volume of the testes was significantly smaller than that of wild-type mice, and there were no germ cells in the testes of the mice, no sperm in the epididymis, and no offspring could be obtained after mating with female mice. Therefore, Ddx5 gene-deficient male mice have the advantages of stability and complete suppression of spermatogenic function, and are a good research model for male fertility disorders.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Ddx5 gene-deleted spermatogenesis disorder mouse model and a construction method thereof. Background technique [0002] With the development of modern society and economy, the change of people's life style and the deterioration of natural environment, the incidence of infertility is gradually increasing. At present, about 15% of married couples in the world are infertile. Among them, male infertility has become a common sexual medical disease in clinic. Researching and establishing animal models for male infertility research is of great significance for ascertaining the etiology, pathogenesis and drug development of the disease. [0003] At present, methods for establishing spermatogenesis disorder models include physical methods, such as thermal effects and microwave radiation methods; chemical methods, such as tripterygium glycosides, busulfan, estrogen, etc.; immune...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/90A01K67/027
CPCA01K67/0275C12N15/907C12N9/22A01K2217/075A01K2227/105A01K2267/03
Inventor 姚红杰夏晴王丽莎王秀芹
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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