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Application of histone methylation H3K4me3 in mouse ovarian development

A technology for histone methylation and ovarian development, which is used in medical preparations containing active ingredients, drug combinations, compound screening/testing, etc.

Active Publication Date: 2020-10-20
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, the effect of H3K4me3 on mouse ovarian follicle development has not been reported

Method used

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  • Application of histone methylation H3K4me3 in mouse ovarian development
  • Application of histone methylation H3K4me3 in mouse ovarian development
  • Application of histone methylation H3K4me3 in mouse ovarian development

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] To study the optimal concentration and effect of exogenous injection of H3K4me3 inhibitor (BCl-121, SIGMA, product number SML1817, hereinafter named Inhibitor) and H3K4me3 agonist (PBIT, SIGMA, product number SML1058, hereinafter named Agonist) in mice 36 C57BL / J6 female mice aged 21 to 28 days were divided into 3 groups (NC group, Inhibitor group and Agonist group). 24mM and 30mM concentration of drugs BCl-121 and PBIT neck subcutaneous injection (one injection volume is 0.1mL / kg), the control group NC group, is the solvent DMSO of BCl-121 and PBIT drugs (Guangzhou Dingguo Biotechnology Co., Ltd., Product number DH105-9, hereinafter named NC), 2 mice for each concentration were slaughtered 48 hours after exogenous injection of the drug, and the H3K4me3 content in the mouse ovary was detected by Western Blot.

[0046] The results showed that exogenous injection of Inhibitor or Agonist drugs can change the degree of H3K4me3 in mice, and the degree of methylation is relat...

Embodiment 2

[0054] To study the effects of H3K4me3 on the secretion of gonadal axis hormones in mouse blood and the morphology of mouse ovaries, 36 28-day-old C57BL / J6 female mice were divided into 3 groups (NC, Inhibitor and Agonist groups), Inhibitor group and Agonist group The drug BCl-121 and PBIT with a concentration of 6 mM were injected intraperitoneally for 21 consecutive days (one injection volume was 0.1 mL / kg), and 4 mice per group were slaughtered every 7 days after the injection, and the gonadal axis hormone in the mouse blood was detected by ELISA The contents of GnRH, E2, LH and FSH, and the morphology of ovarian follicles of mice after drug injection were observed by HE staining.

[0055] The results showed that 1) inhibiting the level of H3K4me3 in mice can reduce the content of GnRH in the blood of 42-day-old mice ( figure 2 a) Inhibition of the level of H3K4me3 in mice can reduce the content of LH in the blood of 42-day-old mice, and the 49-day-old mice have matured, a...

Embodiment 3

[0102] In order to study the effect of H3K4me3 level on the distribution of H3K4me3 protein in mouse ovary and the apoptosis of mouse ovarian follicles, 36 C57BL / J6 female mice aged 21-28 days were divided into 3 groups (NC group, Inhibitor group and Agonist group). ), the Inhibitor group and the Agonist group received exogenous intraperitoneal injection (0.1 mL / kg) of BCl-121 and PBIT at a concentration of 6 mM for 21 consecutive days. The distribution of H3K4me3 in ovarian follicles of 49-day-old mice was observed by immunohistochemistry, and the apoptosis of ovarian follicles of 49-day-old mice was detected by TUNEL immunofluorescence.

[0103] The results showed that: 1) the level of H3K4me3 in mice decreased, and the H3K4me3 protein distributed on mouse ovarian granulosa cells decreased; the level of H3K4me3 in mice increased, and the H3K4me3 protein distributed on mouse ovarian granulosa cells increased ( Figure 4 ). 2) The level of H3K4me3 in mice decreases, and the n...

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Abstract

The invention discloses an application of histone methylation H3K4me3 in the mouse ovarian development. The effect of H3K4me3 on the mouse ovarian follicle development is studied from the living bodylevel and protein level, and the result confirms that the H3K4me3 degree increase can promote the mouse ovarian follicle development, inhibit the mouse follicle apoptosis, and increase the litter sizeand litter weight of mice; and the H3K4me3 degree lowering before sexual maturity in mice can inhibit the secretion of GnRH and gonadotropin-releasing hormone LH and FSH in the blood of the mice. A foundation for further exploring the mechanism of H3K4me3 targeting and regulating key genes of the mouse ovarian follicle development at the molecular level, and then regulating the mechanism of the ovarian follicle development is laid.

Description

technical field [0001] The invention relates to the field of cell engineering and genetic engineering, in particular to the application of histone methylation H3K4me3 in mouse ovary development. Background technique [0002] The ovary is the basis of the female mouse's reproduction, and its main function is to produce and excrete eggs. The mouse follicle is round and located in the ovarian cortex, which is the constituent unit of the ovary, and its growth and differentiation is an important guarantee for ovarian function. According to the growth status of oocytes and granulosa cells in different stages, follicles can be divided into primordial follicles, primary follicles, secondary follicles, tertiary follicles and mature follicles. Because the granulosa cells in primordial follicles, primary follicles and secondary follicles divide rapidly, and the growth and development speed of these three stages of follicles is relatively fast, they are collectively called growing foll...

Claims

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Application Information

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IPC IPC(8): A61K49/00A61K31/428A61K31/445A61P15/08A61P15/00
CPCA61K31/428A61K31/445A61K49/0008A61P15/00A61P15/08
Inventor 袁晓龙何颖婷张哲李加琪张豪陈赞谋
Owner SOUTH CHINA AGRI UNIV
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