Temporal And Spatial Expression Of Slit2/Robo1Gene Which Effect On The Development Of Prehierarchical Follicles On The Chicken

The laying performance of hens is closely associated with the follicular development whichhas a strict priority characteristic, and visible follicles can be divided into prehierarchicalfollicles with the diameter of1-8mm and hierarchical follicles with the diameter above9mm.Slit protein which study in this thesis is a class of known secreted protein, combined with itsreceptor Roundabout (Robo) acting on the nervous system, ovarian tissue and white blood cellsand so on.This paper is divided into three experiments, the first experiment-to assay the expression ofRobo1gene and Slit2gene in different diameters ovarian of Hy-Line Brown hens between150days (early egg period) and210days (high-yield period) by semi-quantitative RT-PCR. Theresults showed that the level of relative expression of Slit2gene mRNA was higher than Robo1gene mRNA, and the level of relative expression of Slit2gene mRNA in high-yield period washigher than that in early egg period. Large differences in gene Robo1spectrum, in vivo Robo1may be combined with more ligands, having different functions together in the follicle, and thatwould affect the laying performance.The second experiment-by Enzyme immunohistochemical techniques, To assay theexpression of Slit2protein and Robo1protein in different diameter follicle of Hy-Line Brownhens including30d、60d、120d and150d and found that both Slit2and Robo1protein expressedin different age of Hy-Line Brown hens ovaries, and all expressed in follical oocytes andgranulosa cells, but did not express in the theca cells. Slit2protein and Robo1protein positivesignals showed different colored shades through immunohistochemical sections and found thatthere are no significant difference in protein expression in ovarian follicles at different ages forSlit2protein and Robo1protein(P>0.05), but the level of Slit2protein expression was relativelyhigher than that of Robo1protein. These results suggest that Slit2protein and Robo1protein allwere expressed in ovarian development of different ages, and these proteins might throughautocrine or paracrine involve in follicular oocytes growth and formation and migration ofgranulosa cell, thus affecting follicular development.The third experiment-the preliminary experiment used different concentrations of FSThormone and FSH hormone to treat prehierarchical follicles (diameter5-8mm), and found that the process-time-48hours and deal with the concentration-10ng/ml had a significant effect onthe expression level of Slit2genes and Robo1gene mRNA. Using10ng/ml of FST hormone,FSH hormone, FSH+FST mixed hormones and the control group respectively to treat fourprehierarchical follicles including diameter of4-5mm, diameter of5-6mm, diameter of6-7mmand diameter of7-8mm, three genes treated by FST+FSH, the diameter of7-8mm follicles havethe highest relative expression levels, Slit2/Robo1highly expressed in this developmental stage,may be involved in the choice of the follicle. In addition, FSH+FST treatment and control, thepurpose of changing the trend to the relative expression of the three genes is consistent withincreasing the diameter of the follicles, and FSH+FST treatment relative expression levelhigher than the control relative expression levels, indicating that in vitro FSH hormones andFST hormones may be some synergy, but the exact mechanism is unclear, remains to be furtherstudied.