褪黑激素RIA方法的建立及其在探索湖羊四季发情机理上的应用  

Development of a RIA for Melatonin and Its Application to the Research on the Oestrus Mechanism of Out-of-season in Hu Sheep

作　　者：汪兴生[1] 夏银 尚平[3] 陈伟华[4] 钱建共 程瑞禾[4] 何良军[4] 张勇[4] 沈子龙[1] 

机构地区：[1]中国药科大学生物技术中心,江苏南京210009 [2]UniversityofNewEngland(Australia [3]南京军区总医院 [4]南京农业大学 [5]苏州畜牧兽医站

出　　处：《中国兽医学报》2001年第3期287-290,共4页Chinese Journal of Veterinary Science

基　　金：江苏省自然科学基金!资助项目 ( BK970 95 )

摘　　要：运用 Mannich反应 ,将褪黑激素 ( MLT)与牛血清白蛋白 ( BSA)连结 ,制备 MLT完全抗原。 MLT与BSA连结比为 1 2∶ 1。将此完全抗原免疫 5只雄兔 ,制备特异性抗体 ;将 MLT用 12 5I标记 ,薄层层析筛选标记物 ,制备高纯度的放射性标记物 ,建立竞争性 MLT-RIA法。本法可测范围为 1 5.3 1～ 1 50 0 pg,灵敏度为每管 1 5.3 1 pg,批内变异系数 6.7% ,批间变异系数 1 4 .8%。用此法测定夏至时考力代羊昼夜血样 ,测定结果与国外基本一致 ;测定夏至时湖羊昼夜血样 ,发现湖羊 MLT分泌有其特殊性 ,即血液 MLT水平呈明显的昼夜变化 :白天为 1 0 0 ng/L左右 ,而黑暗开始时 ,MLT水平迅速上升 ,在 2 0 :0 0～次日 8:0 0处于高水平 ,最大值为 1 1 0 0 ng/L左右。A RIA for determination of melatonin(MLT) in animal blood was described.The rabbit antibody employed was raised to the conjugate formed by coupling melatonin to bovine serum albumin with formaldehyde reaction(MLT∶BSA 12∶1).MLT was successfully conjugated with 125 I and MLT I 125 was purifed by TLC.Finally the standard curve was created.The sensitivity of the method was 15.31 pg/tube.The variation coefficiences of intra and inter plates were 6.7% and 14.8%,respectively.The MLT concentration of 24 h blood sample of corridale sheep at summer solstice was tested by the method to be similar to that in litteratures and different from the other sheep.The results showed that the serum MLT level of Hu Sheep at summer solstice revealed a nyctohemeral fluctuation with the peak of 1 100 ng/L at 20:00~8:00.

关 键 词：褪黑激素 RIA 湖羊 回季发情机制 神经激素 分泌特性 

分 类 号：Q576[生物学—生物化学] S827.8[农业科学—畜牧学]