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广西沼泽型水牛垂体中促性腺激素释放激素的免疫组化定位及其受体基因表达
引用本文:李瑞明,凌泽继,宋小白,秦津,谢莹雪,史玉静.广西沼泽型水牛垂体中促性腺激素释放激素的免疫组化定位及其受体基因表达[J].安徽农业科学,2010,38(24):13164-13166,13174.
作者姓名:李瑞明  凌泽继  宋小白  秦津  谢莹雪  史玉静
作者单位:1. 广西大学动物科学技术学院,广西南宁,540003
2. 扬州大学兽医学院,江苏扬州,225009
基金项目:硕士研究生科研创新项目,广西自然科学基金,广西教育厅科研基金,广西大型仪器协作共用网资助 
摘    要:目的]探讨广西沼泽型水牛垂体中促性腺激素释放激素(GnRH)的细胞定位以及是否存在促性腺激素释放激素受体(GnRH—R)。为GnRH-R的克隆及其序列分析提供理论依据。方法]采用免疫组织化学ABC法,确定促性腺激素释放激素在垂体中的定位,结合图像分析系统检测GnRH在垂体中的含量,并利用RT—PCR技术扩增其受体基因片段。结果]腺垂体远侧部中呈现较强的阳性GnRH免疫反应,阳性物质主要分布在胞质,胞核呈阴性。神经垂体中无阳性信号。同时,在垂体中扩增出大小为1008bp的GnRH—R基因片段。结论]腺垂体远侧部边缘区中有大量GnRH及其受体存在,证实垂体前叶是下丘脑-垂体-性腺轴以及多个内分泌轴相互作用的重要部位。

关 键 词:促性腺激素释放激素  促性腺激素释放激素受体  免疫组化  RT-PCR  广西沼泽型水牛  垂体

Immunohistochemical Localization and Receptor Gene Expression of Gonadotropin Releasing Hormone in Hypophysis of Guangxi Swamp Buffaloes
Institution:LI Rui-ming et al ( College of Animal Science and Technology, GuangXi University, Nanning, Guangxi 540003 )
Abstract: Objective ] To explore the cellular location of gonadotropin-releasing hormone (GnRH), detect the gonadotropin-releasing hor- mone receptor (GnRH-R) in hypophysis of Guangxi swamp buffaloes and provide a theoretical basis for cloning and sequence analysis of GnRH-R gene. Method] The location and expression of GnRH in hypophysis of Guangxi swamp buffaloes were determined by using immunohistoehemical ABC method and image analysis. The GnRH-R gene fragment was amplified by real-time PCR. Result] Many positive GnRH cells were detected in adenohypophysis, the positive substances were mainly distributed in cytoplasm and it was negative in nuclei. There was no positive signal in neurohyppohsis. Meanwhile, the GnRH-R gene fragment was amplified with 1 008 bp in size. Conclusion] A large number of GnRH and GnRH-R exist in the pars distalis of adenohypophysis, which indicates that the anterior hypophysis plays an important role in the hypothalamus-pituitary-gonadal (HPG) axis and the interaction of multiple endocrine axes.
Keywords:RT-PCR
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