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Application of mitochondrion protein translation factor Guf1 in male sterile research

A male sterility and male technology, which is applied in the direction of introducing foreign genetic material using carriers, veterinary instruments, and microbial measurement/inspection, etc. It can solve problems such as membrane potential decline, affecting tissue and organ functions, and reducing ATP production.

Active Publication Date: 2014-03-05
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, research on mitochondrial respiratory function and respiratory chain energy metabolism at home and abroad is mainly focused on some tissues and organs with high oxygen consumption and high energy demand, such as liver tissue, brain tissue, myocardial tissue, etc. Mitochondrial energy metabolism disorders in these high oxygen consumption tissues It will cause a decrease in membrane potential, greatly reduce the generation of ATP, and affect the function of tissues and organs

Method used

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  • Application of mitochondrion protein translation factor Guf1 in male sterile research
  • Application of mitochondrion protein translation factor Guf1 in male sterile research
  • Application of mitochondrion protein translation factor Guf1 in male sterile research

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1, Guf1 gene knockout strategy and PCR, Western blot detection.

[0027] In order to study the function of Guf1 protein in higher organisms and what impact its mutation will have on the body. We chose mice as a model organism, used gene targeting technology to knock out Guf1 protein, and used PCR and Western blot methods to evaluate the knockout efficiency. figure 1 The strategy for Guf1 protein knockdown and the knockout efficiency were tested by PCR and western blotting. figure 1 A shows the knockdown strategy of Guf1 figure 1 B represents the genotype of Guf1 knockout mice identified by PCR. figure 1 B represents the knockout efficiency of Guf1 in mouse tissues detected by western blot. The results showed that the amount of Guf1 protein was significantly reduced in the testis, further evidence that the protein was knocked out in mice.

[0028] specific method:

[0029] Obtaining Guf1 knockout mice:

[0030] Construction of the Guf1 targeting vector: the...

Embodiment 2

[0048] Example 2. Pathological analysis of male sterility caused by Guf1 gene knockout.

[0049] In order to further study the dysfunction caused by the loss of Guf1, we focused on the pathological study of the reproduction of Guf1 knockout mice, including the number of offspring, sperm motility, and pathological sections and transmission electron microscope analysis of testes. figure 2 A represents the analysis of the number of offspring, which shows that the male knockout mice and normal female mice have mating behavior but no offspring. figure 2 B Statistics of the sperm counts of wild-type and Guf1-knockout mice showed that the number of sperm in Guf1-knockout mice was significantly reduced. figure 2 C is the observation of testis morphology, and it was found that the testis morphology of Guf1 knockout mice became smaller. figure 2 D is the observation of sperm morphology by confocal fluorescence microscope, and it is found that the mitochondrial sheath of the knockou...

Embodiment 3

[0054] Example 3. Guf1 knockout increases sperm cell apoptosis and affects spermatogenesis.

[0055] In order to study the effect of Guf1 on the process of mouse spermatogenesis, we stained the reproductive organs of wild-type and Guf1-knockout male mice, including testes and epididymis, and analyzed sperm motility. image 3 A and C are the H&E staining of the knockout epididymis and testis, which shows that there are a large number of immature sperm cells in the knockout mouse epididymis. image 3 B is TUNEL detection of apoptosis in testis and epididymis, indicating that Guf1 knockout will produce obvious apoptosis. image 3 D and E are the analysis of sperm motility by computer assisted sperm analyzer (CASA), and the results show that the sperm motility of knockout mice is significantly reduced. Through the above pathological analysis of Guf1 knockout, it was found that Guf1 knockout would cause spermatogenesis disorder and lead to male sterility.

[0056] specific method...

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Abstract

The invention provides application of a model organism mouse with the Guf1 gene knocked out in the male sterile mechanism, and particularly provides application of the mouse model with the Guf1 gene knocked out in human male reproduction and spermatogenesis process research and male sterile mechanism research and treatment, and the application can become a diagnostic method for diagnosing male sterility.

Description

technical field [0001] The invention relates to the application of the mitochondrial protein translation factor Guf1 in the research of male reproduction. Specifically, the present invention provides the use of a model organism mouse that knocks out the Guf1 gene in the study of male sterility. More specifically, the present invention provides a mouse model for the knockout of the Guf1 gene in the study of human male reproduction and spermatogenesis. and uses in the mechanisms and treatment of male infertility. Background technique [0002] At present, about 15% of married couples in the world are troubled by infertility, and asthenospermia accounts for 19% of male infertility, which has become a common cause of male fertility. The energy required for sperm motility mainly comes from the oxidative phosphorylation of the mitochondrial respiratory chain. If the function of oxidative phosphorylation is abnormal, the change of enzyme activity or expression level and the variat...

Claims

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Application Information

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IPC IPC(8): A01K67/027C12Q1/68C12N15/85A61D19/04
Inventor 秦燕高岩岩
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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