【作者】 李珣； 雷治海； 苏娟； 赵洋阳； 靳蒙蒙； 房蕊； 郑路程； 焦洋；

【Author】 Xun Li,Juan Su,Zhihai Lei~*,Yangyang Zhao,Mengmeng Jin,Rui Fang,Lucheng Zheng,Yang Jiao (College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,P.R.China)

【机构】 南京农业大学动物医学院；

【摘要】 (目的)自从GnIH发现以来被认为是动物繁殖调控中关键的神经肽。GnIH通过其受体GPR147抑制促性腺激素的合成和释放。为了研究猪GnIH的生理功能,(方法)本研究克隆GnIH前体肽和GPR147并测序。然后通过荧光定量PCR研究它们在各组织中及发情周期中在下丘脑-垂体-卵巢轴的表达情况。利用免疫组化也研究了GnIH及GPR147在发情周期卵巢中的分布定位。(结果)结果显示克隆的猪GnIH前体肽能编码三个LPXRF,受体具有典型的跨膜结构。组织表达结果显示GnIH主要在脑中表达而受体的表达与之相似。发情周期中在生殖轴的表达结果显示GnIH及GPR147均在发情期表达最低,分别在发情前期和间情期表达最高。在垂体中两者均在发情后期表达量最高然而在卵巢中GnIH与GPR147表达模式似乎相反。免疫组化结果显示发情前期、发情期中GnIH、GPR147免疫阳性反应出现在囊状卵泡的卵巢颗粒细胞及卵巢膜细胞而在发情后期、间情期则出现在黄体细胞。(结论)因此,本研究为深入探讨猪GnIH的生理功能提供分子生物学及形态学理论依据。更多还原

【Abstract】 (Objective) Since its discovery,gonadotropin-inhibitory hormone(GnIH) has appeared to act as a key neuropeptide in the control of vertebrate reproduction.GnIH acts via the novel G protein-coupled receptor 147(GPR147) to inhibit gonadotropin release and synthesis.To determine the physiological functions of GnIH in the pig,(Method) a study was conducted to clone and sequence the cDNA of the GnIH precursor and GPR147.Subsequently,tissue expression studies and the expression patterns in the reproductive axis of the female pig across the estrous cycle were also systemically investigated by real-time PCR.Distribution of GnIH and GPR147 in the ovary during the estrous cycle was studied by immunohistochemistry.(Result) Our results demonstrated that the cloned pig GnIH precursor cDNA encoded three LPXRF and that its receptor possessed typical transmembrane features.Tissue expression studies revealed that GnIH was mainly expressed in the brain,corresponding largely with the tissue expression patterns of GPR147 in the pig.The expression patterns in the reproductive axis of the female pig across the estrous cycle revealed that the hypothalamic levels of both GnIH and its receptor mRNA were lowest in estrus and peaked in the proestrus and diestrus phases.The highest pituitary GnIH mRNA level was detected in the metestrus,and its receptor displayed a somewhat similar pattern of expression to that of the ligand.However,the expression patterns of GnIH and GPR147 were negatively correlated in the ovary.Immunolocalization in the ovary during the estrous cycle revealed that the immunoreactivities of GnIH and GPR147 were mainly localized in the granulosa and theca cells of the antral follicles during proestrus and estrus and in the luteal cells during metestrus and diestrus.(Conclusion) Taken together,this research provided molecular and morphological data for further study of GnIH in the pig.更多还原