Cloning Of Goose GnIH And Its Receptor, The Gene Expression And Research In Granulosa Cells

gonadotropin-inhibiting hormone (GnIH) could decrease gonadotripin release of quail anterior pituitary in a dose-dependent manner. Meanwhile, it is also the only neuropeptide to inhibit the development and function of reproductive system. It could regulate the reproduction by hypothalamus, pituitary gland and gonad. In addition to breeding, it also plays an important role in stress response, food intake and seasonal breeding, etc. To study the effects of GnIH on goose breeding, we has detected the expression pattern of GnIH and its two receptors RFRPR and NPFFR in laying goose different tissues, and then choose the goose ovarian granulosa cells as the research platform, we study the molecular mechanisms by adding different concentrations of exogenous GnIH treatment.The main findings are as follows:(1) The partial CDS sequence of GnIH, RFRPR and NPFFR has been amplified. We found that the homology of nucleotide sequence between RFRPR and NPFFR is 63.35%, and both of they have 7 similar amino acid transmembrane domain.(2) We analyze the gene exprssion pattern in laying goose. The highest expression of GnIH were found in the hypothalamus and the adrenal gland, ovaries and fallopian tubes also detected expressed. This means the hypothalamus is the mainly sites of GnIH synthesis.(3) The RFRPR and NPFFR were mainly expressed in the brain, and the RFRPR is more in line with the distribution of immune nerve fibers of GnIH, that suggest RFRPR is the main receptor of GnIH, And RFRPR also expressed in the adrenal glands, eye and ovary, it means that GnIH may be play roles in stress response, seasonal breeding and directly regulate the ovarian functions. And in brain(4) We use the cultured goose ovarian granulosa cells as our template of study, and use different dose of GnIH (0,0.1,10,1000 ng/ml) to treat the cells. The results found that GnIH could directly affect the synthesis and secretion of steroid hormones in granulosa cells. The experimental results indicate that GnIH could decrease the exprssion of StAR,3/3-HSD and CYP19 in cultured granulosa cell, and decline the secretion of E2 and P4.(5) Exogenous GnIH could reduce the cell viability of granulosa cells, promote apoptosis and reduce proliferation. Furthermore, GnIH treatment can reduce the expression of cell CCND1, CCND2 and Bcl-2 gene.(6) After GnIH treated, GnIH could reduce the expression of LH-R and FSH-R, but not the GnRH and RFRPR. This indicating that GnIH may be regulate the granulosa cell through LH-R and FSH-R...