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当前位置: 首页 > 文章 > 鲟鱼GtH放射免疫测定方法的建立及催产前后史氏鲟血清GtH含量的变化 水产学报 2008,32 (6) 3-10
Position: Home > Articles > Setting up radioimmunoassay for sturgeon gonadotropin GtH and assay of Amur sturgeon serum GtH after inducing ovulation Journal of Fisheries of China 2008,32 (6) 3-10

鲟鱼GtH放射免疫测定方法的建立及催产前后史氏鲟血清GtH含量的变化

作  者：
胡红霞;刘晓春;朱华;张勇
单  位：
中山大学有害生物控制与资源利用国家重点实验室;国家淡水渔业工程技术研究中心(北京)暨北京市水产科学研究所
关键词：
史氏鲟;俄罗斯鲟;促性腺激素;放射免疫测定;催产
摘  要：
用蛋白质快速液相层析(FPLC)分离俄罗斯鲟鱼脑垂体促性腺激素(GtH),共获得GtH纯化蛋白4.6mg。用SDS-PAGE变性胶电泳图谱初步分析纯化蛋白,用反相高效液相色谱(rpHPLC)分离出两个亚基,质谱测定α亚基分子量:15 603ku,两个β亚基分子量分别为14 338ku和14 694ku。制备了鲟鱼GtH的兔抗血清多克隆抗体,采用氯胺T法用125I标记抗原,用羊抗兔γ球蛋白做二抗,建立鲟鱼GtH的放射免疫测定方法。测定了催产前后不同时间史氏鲟血清中GtH含量的变化,统计分析表明成功排卵的雌鱼比未排卵和阴性对照组在催产后12h和16h GtH分泌量升高显著,最高达到234ng.mL-1;催产效应时间与催产后血清中GtH的含量紧密相关。
译  名：
Setting up radioimmunoassay for sturgeon gonadotropin GtH and assay of Amur sturgeon serum GtH after inducing ovulation
作  者：
HU Hong-xia 1,LIU Xiao-chun2,ZHU Hua1,ZHANG Yong2(1.National Freshwater Fisheries Engineering Technology Research Center,Beijing Fisheries Research Institute,Beijing 100068,China;2.Institute of Aquatic Economic Animals,and Guangdong Province Key Laboratory for Aquatic Economic Animals,Sun Yat-sen University,Guangzhou 510275,China)
关键词：
Amur sturgeon;Russian sturgeon;gonadotropin(GtH);radioimmunoassay(RIA);induced ovulation
摘  要：
Gonadotropin(GtH) protein was purified from the pituitary of Russian sturgeon using fast protein liquid chromatograph(FPLC).A total of 4.6 mg of Russian sturgeon GtH in dry weight was obtained.The purified protein was analyzed by SDS-PAGE electrophoresis.Two subunits of GtH were separated by reversed-phase high-performance liquid chromatography(rpHPLC).The molecular weight was determined through mass spectrograph.The α subunit was 15 603 ku and the two β-subunits were 14 338 ku and 14 694 ku respectively.Rabbit anti-sturgeon GtH polyclone antibody was prepared;purified GtH was labeled by 125I using chloramines T as labeled antigen;then the sturgeon GtH radioimmunoassay(RIA) method was set up using the goat anti-rabbit γ-globulin as the second antibody.After inducing ovulation,changes of serum GtH levels were detected.The secretion peak of female fish which ovulated successfully was sustained from 12 to 16 hours after first inducing injection and the serum GtH levels were increased significantly compared with control group and un-ovulated fish.The effect time of LHRH-A was related closely to the serum GtH levels.

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