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半滑舌鳎(Cynoglossus semilaevis)新型膜孕激素受体基因(mPRL)在卵母细胞成熟过程中的表达特征
引用本文:史 宝,柳学周,徐 涛,李晓妮,徐永江,张金勇. 半滑舌鳎(Cynoglossus semilaevis)新型膜孕激素受体基因(mPRL)在卵母细胞成熟过程中的表达特征[J]. 渔业科学进展, 2017, 38(1): 10-17. DOI: 10.11758/yykxjz.20151105003
作者姓名:史 宝  柳学周  徐 涛  李晓妮  徐永江  张金勇
作者单位:1. 农业部海洋渔业可持续发展重点实验室 青岛市海水鱼类种子工程与生物技术重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;青岛海洋科学与技术国家实验室 海洋渔业科学与食物产出过程功能实验室 青岛 266071;2. 山东省渔业技术推广站 济南250013;3. 农业部海洋渔业可持续发展重点实验室 青岛市海水鱼类种子工程与生物技术重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071
基金项目:国家自然科学基金项目(31201982)、国家鲆鲽类产业技术体系(CARS-50)和山东省优秀中青年科学家科研奖励基金项目(BS2013SW042)共同资助
摘    要:为进一步提高半滑舌鳎(Cynoglossus semilaevis)人工育苗技术的水平,对半滑舌鳎新型膜孕激素受体(mPR-like,mPRL)基因的表达特征和作用机制进行了研究.应用实时定量PCR方法分析mPRL mRNA在卵子形成过程中的时序表达,发现半滑舌鳎mPRL mRNA相对表达量的最高值出现在性成熟阶段卵巢的Ⅴ时相卵母细胞.原位杂交分析mPRL mRNA在繁殖相关组织的细胞学定位,发现mPRL mRNA分布在半滑舌鳎性成熟阶段卵巢的卵母细胞膜上;在脑的神经元和垂体内分散的细胞中,mPRL mRNA阳性信号较强.制备半滑舌鳎mPRL的多克隆抗体,采用Westem blotting方法检测半滑舌鳎mPRL蛋白在不同组织的表达特征,发现mPRL蛋白的表达量在卵巢、脑、垂体中相对较高,在肝脏、头肾、肾脏中表达量相对较少.免疫组化结果显示,半滑舌鳎mPRL蛋白在卵巢、脑和垂体的细胞学定位与mPRL mRNA定位一致.应用实时定量PCR和Westem blotting方法检测促性腺激素调控下半滑舌鳎不同时相卵母细胞mPRL基因mRNA和蛋白的表达变化,结果显示,促性腺激素对半滑舌鳎卵母细胞中mPRL基因mRNA和蛋白表达都有一定的上调作用,特别是对Ⅴ时相卵母细胞中mPRL mRNA和蛋白的表达量提升明显;发现表达量与促性腺激素调控作用具有剂量依存关系.半滑舌鳎mPRL在繁殖相关组织的表达特征表明其通过脑-垂体-卵巢轴参与繁殖调控,同时也揭示了mPRL介导卵母细胞成熟机制.

关 键 词:半滑舌鳎  新型膜孕激素受体  卵母细胞成熟  mRNA表达  蛋白表达
收稿时间:2015-11-05
修稿时间:2015-12-02

Expression Characterization of the Novel Membrane Progestin Receptor (mPR-Like) Gene During the Oocyte Maturation of Half-Smooth Tongue Sole (Cynoglossus semilaevis)
SHI Bao,LIU Xuezhou,XU Tao,LI Xiaoni,XU Yongjiang and ZHANG Jinyong. Expression Characterization of the Novel Membrane Progestin Receptor (mPR-Like) Gene During the Oocyte Maturation of Half-Smooth Tongue Sole (Cynoglossus semilaevis)[J]. Progress in Fishery Sciences, 2017, 38(1): 10-17. DOI: 10.11758/yykxjz.20151105003
Authors:SHI Bao  LIU Xuezhou  XU Tao  LI Xiaoni  XU Yongjiang  ZHANG Jinyong
Affiliation:Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071,Shandong Province Fisheries Technology Extension Station, Jinan 250013,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;,Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266071 and Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;
Abstract:
Keywords:Cynoglossus semilaevis   mPR-Like   Oocyte maturation   mRNA expression   Protein expression
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