The Research Of Slit3/Robo2Gene Expression Patterns In Prehierarchical Follicles And The Impact Of ActivinA Of Chicken

Slit3/Robo2was first found in nerve cells, belong to Nerve guide factors. Recent studieshave found the expression of Slit3/Robo2gene in mammalian reproductive organs such asovarian and endometrial, indicating that Slit3/Robo2may have a close relationship withfollicular development in animals. ActivinA is a member of transforming growth factor β(TGF-β) superfamily, is widely distributed in the ovaries and testes, play a regulatory functionmainly through the pituitary-gonadal axis in a variety of tissue cells. Studies have shown thatActivinA can stimulate the proliferation of granulosa cells and promote the development offollicles. In this study, immunohistochemistry, semi-quantitative RT-PCR and follicle culturemethods on the ovaries Slit3/Robo2gene expression patterns and Activin A study its impact.Thisexperiment by immunohistochemical, semi-quantitative RT-PCR and follicle culture methods tostudy the expression patterns of time and space of Slit3/Robo2gene in the ovary and the impactof ActivinA.In this study, the protein localization analysis of ActivinβA, Silt3, Robo2and Foxl2gene atdifferent levels of development follicles in the ovaries Hy-Line Brown hens are detected byimmunohistochemical methods. The results showed that in oocytes and granulosa cells candetecte ActivinβA, Silt3, Robo2and Foxl2gene positive signal, there are no positive signals ofSilt3, Robo2gene in follicle cell membrane layer, while ActivinβAgene positive signal wasdetected in primary follicle cell membrane layer, but positive signals were not detected inPrehierarchical follicles cell membrane layer.The expression levels of ActivinβA, Silt3, Robo2and Foxl2gene mRNA in150days and210days of age at different developmental levels of ovarian follicles Hy-Line Brown hens weredetected by semi-quantitative RT-PCR method.The results showed that: ActivinβA, Silt3, Robo2and Foxl2genes are expressed and have a high level of expression in each grade follicles, theexpression of ActivinβAwas gradually increased as the follicle development,with a highexpression level in F1and compared with other grade follicles was difference significantly(P<0.05); interstitial,1-4mm,4-5mm and other follicles were no significant differences (P>0.05).Silt3/Robo2gene exist in the form of match receptors; the expression quantity in each gradefollicles was almost the same change trend. Foxl2gene has a higher expression, the expressionquantities of Foxl2gene in150days of age at different developmental levels of ovarian folliclesHy-Line Brown hens were not significant (P>0.05), the expression quantities of Foxl2gene in 210days of age at different developmental levels of ovarian follicles Hy-Line Brown hensbetween F6and other grade follicles were different significantly (P<0.05).Using different concentrations of Activin A and InhibinA cultured follicles of the same size.The results showed that when the concentration of the hormone10ng/ml, cultured for24hours,with the best results. On this basis, we use the concentration of hormones on different sizedfollicles are processed, the results showed that follicles with ActivinA and InhibinA cultured,which Silt3, Robo2and Foxl2gene expression levels compared with the control group theincrease and had a significant difference (P<0.05), in the same size of follicles, each gene wasdifferent hormones have the same trend. Thus, we can determine Silt3, Robo2and Foxl2genefollicles may play a catalytic role while speculation ActivinA has a certain influence on thisprocess.