Target protein for improving spermatogensis

A technology of target protein and spermatogenesis, applied in the field of target protein for improving spermatogenesis, can solve the problems of decreased fertility, decreased sperm count and decreased sperm motility in male mice

Inactive Publication Date: 2013-09-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In 2007, Duprey et al. studied the expression characteristics of βB2 crystallin in mouse testis: when the mouse was just born, βB2 crystallin was expressed in the testis, but the expression level was low. As the mouse matured, its expression The amount gradually increased, maintaining a high level of expression in the mature testis; and it was found that the Swiss Webster male mice with βB2 crystal protein gene mutations had reduced fertility, which manifested as a decrease in the number of sperm in the testis and a decrease in sperm motilit

Method used

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  • Target protein for improving spermatogensis
  • Target protein for improving spermatogensis
  • Target protein for improving spermatogensis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0532] Example 1, βB 2 Establishment and Identification of Crystallin Knockout Mice Materials and Methods

[0533] 1. Experimental animals

[0534] The animals used in the experiments were mice of C57BL / C strain.

[0535] 2. Experimental materials

[0536] (1) Main reagents and instrument consumables

[0537]

[0538]

[0539] (2) Preparation of main solutions and reagents

[0540] 1. Electrophoresis buffer TBE

[0541] Weigh 2.42g of Tris-Base, 2.75g of boric acid, 1ml of 0.5M EDTA, add double distilled water to make up to 500ml.

[0542] 2. Genome extraction kit related solutions

[0543] Prepare your own absolute ethanol. In the DNA extraction kit, buffer GD and rinse solution PW are prepared by adding absolute ethanol as required before use.

[0544] 3. 0.7~1% agarose gel solution

[0545] Weigh 0.5-0.6 g of agarose, put it into an Erlenmeyer flask, add 80-90 ml of TBE electrophoresis buffer, and dissolve to form an agarose gel solution with a concentration o...

Embodiment 2

[0600] Example 2, βB 2 Effects of Crystallin Gene Knockout on the Reproductive Function of Male MiceMaterials and Methods

[0601] 1. Experimental materials

[0602] (1) Main reagents

[0603]

[0604]

[0605] (2) Main instruments and consumables

[0606]

[0607] (3) Preparation of main solutions and reagents

[0608] 1. Preparation of related reagents for Western blotting

[0609] (1) 4× protein loading buffer: 5ml of 1M Tris-HCl (pH6.8), 0.8g of SDS, 2ml of glycerol, 0.0012g of bromophenol blue, 0.4ml of β-mercaptoethanol, and dilute to 10ml with deionized water.

[0610] (2) 1.5M Tris-HCl (pH8.8): Weigh 18.671g of Tris, dissolve in 100ml of distilled water, and adjust the pH to 8.8 with concentrated hydrochloric acid.

[0611] (3) 1M Tris-HCl (pH6.8): Weigh 12.114g of Tris, dissolve in 100ml of distilled water, and adjust the pH to 6.8 with concentrated hydrochloric acid.

[0612] (4) 10% ammonium persulfate (abbreviation: AP): 1g of ammonium persulfate pow...

Embodiment 3

[0749] Example 3, βB2 Crystallin Gene Knockout Causes Mechanism of Reduction of Male Mice Reproductive Function Materials and Methods

[0750] 1. Experimental materials

[0751] (1) Main reagents and instrument consumables

[0752]

[0753] (2) Preparation of main solutions and reagents

[0754] 1. Liquid preparation related to TUNEL detection

[0755] TUNEL detection blocking solution: 3% hydrogen peroxide dissolved in methanol, with 30% H 2 o 2 And 80% methanol solution preparation.

[0756] TUNEL detection permeation solution: 0.1% TritionX-100 dissolved in 0.1% sodium citrate.

[0757] TUNEL detection DNase reaction solution: 100~3000DNase, 40mM Tris-HCL PH7.9, 10mM NaCL, 6mM MgCL2, 10mM CaCL 2 .

[0758] TdT enzyme reaction solution: 45ul equilibration buffer (Equilibration Buffer) + 1ul Biotin-11-dUTP + 4ul TdT Enzyme mixed.

[0759] Streptavidin-HRP working solution: mix 0.5ul Streptavidin-HRP+99.5ul PBS.

[0760] 2. BrdU dissolution

[0761] Dissolve BrdU ...

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Abstract

The invention relates to a target protein which takes a betaB2 crystal protein to improve the spermatogensis and a novel application thereof. A betaB2 crystal protein knockout mouse model is used for analyzing the expression of the betaB2 crystal protein in mouse testis tissues to confirm the reduction of CaMKIV expression and activity, caused by betaB2 crystal protein knockout, so as to reduce a reproductive function; and a CaMKIV expression level is reduced so that the expression of Bcl-2 is reduced to enable testis germ cells to be excessively proliferated and apoptotic and damage sperms. Therefore, the target protein for improving the spermatogensis is the betaB2 crystal protein, and a novel medicine resource and a novel application preparation are provided; and the target protein has the very important value for improving the existing medical level. The target protein disclosed by the invention is safe and cheap, effective in treatment and stable in property, is suitable for large-scale production and business application of industries of medicines, reagents and the like, is easy to popularize and apply, has a very good application prospect, and has obvious social benefits and economic benefits.

Description

technical field [0001] The present invention relates to the technical fields of medicine, food, reagents, etc., specifically to a new application of crystal protein, more specifically to a target protein for improving spermatogenesis and its new application, and more specifically Said to be a crystal protein involving βB2 (or: βB 2 , betaB2, crybb 2 or crybb2) can be used as a target protein for improving spermatogenesis and its new application. Background technique [0002] βB 2 Advances in research on the functions of crystallins in and out of the lens [0003] Crystallins are usually expressed in the lens of mammalian eyes. They are the main structural proteins in the cytoplasm of the mammalian lens and play an important role in maintaining the lens' ability to refract and transmit light. According to its migration ability in the electric field, it can be divided into three categories: α, β, and γ (Graw J. Genetics of crystallins: cataract and beyond. Exp Eye Res, 200...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P15/08A23L1/305
Inventor 李闻捷邓安梅相芬芬高谦高莉张俊洁
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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