拟穴青蟹GnRH-R的免疫识别  

Immunological Recognition of Gonadotropin-releasing Hormone Receptor(GnRH-R) in the Mud Crab,Scylla paramamosain

作　　者：陶永[1] 刘元婧[1] 叶海辉[1] 王桂忠[1] 李少菁[1] 

机构地区：[1]厦门大学海洋与环境学院,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2011年第5期799-802,共4页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金项目(41076081)

摘　　要：促性腺激素释放激素（GnRH）及其受体（GnRH-R）对生殖具有重要的调控作用．迄今无脊椎动物GnRH及GnRH-R的研究尚少．本研究采用兔抗人GnRH-R、兔抗果蝇GnRH-R和兔抗海鞘GnRH-R的抗体，应用免疫印迹和免疫共沉淀技术对拟穴青蟹（Scylla paramamosain）GnRH-R进行了免疫识别，所得结果如下：1）经免疫印迹检测，拟穴青蟹脑、胸神经团、视神经节和精巢中共有的免疫阳性条带分子质量为45-55ku．脑和胸神经团中另有一条36ku条带；在精巢中，兔抗人GnRH-R和兔抗果蝇GnRH-R抗体只有一条28ku条带，而兔抗海鞘GnRH-R抗体显示有28，36和30ku的条带．2）利用免疫共沉淀技术，分离出与GnRH-R抗体相结合的两种物质，分子质量分别为38．1和54．0ku，分别与哺乳动物和非哺乳动物GnRH-R的分子质量接近．该发现提示拟穴青蟹体内存在GnRH-R，为深入了解甲壳动物生殖调控机理提供了理论依据．Gonadotropin-releasing hormone （GnRH） and its receptor （GnRH-R）play an important role in the regulation of reproduction. To date,there has been little study on the GnRH and GnRH-R in invertebrates. In this study,with three GnRH-Rs of rabbit antibody against different species, including human, fruit fly and sea squirt, Western blotting and co-immunoprecipitation were applied to test the GnRH-R in mud crab, Scylla paramamosain. The common immunoreactive bands, with the molecular mass between 45 and 55 ku, appeared in the brain, thoracic ganglion,eyetalk and testes. Another band with 36 ku was found in the brain and thoracic ganglion. And in the testes, there was only a band with 28 ku using the antibody against human and fruit fly, while using rabbit antibody against sea squirt, there were bands with 28,36 and 30 ku. Two substance combined to GnRH-R were separated by eo-immunoprecipitation,with the molecular mass of 38. 1 and 54.0 ku, which were close to the mammal and non-mammal, respectively. This study shows that GnRH-R occurs in mud crab, which is helpful to further understanding of the regulation of reproduction in crustaceans.

关 键 词：GnRH-R 甲壳动物 免疫印迹 免疫共沉淀 

分 类 号：R446.5[医药卫生—诊断学]