减数分裂期雄性大菱鲆孕激素及受体基因的表达分析  

Progestin actions and expression analysis of progestin receptors in the meiotic in male turbot(Scophthalmus maximus)

作　　者：丰程程 柳意樊[1,3] 安皓 徐世宏[1,3] 王彦丰 宋宗诚[4] 刘清华 李军[1,3] 

机构地区：[1]中国科学院海洋研究所中国科学院实验海洋生物学重点实验室,山东青岛266071 [2]中国科学院大学,北京100049 [3]青岛海洋科学与技术国家实验室海洋生物学与生物技术功能实验室,山东青岛266237 [4]山东威海圣航水产科技有限公司,山东威海264200

出　　处：《海洋科学》2017年第8期91-98,共8页Marine Sciences

基　　金：国家自然科学基金资助项目(31472264;31572602);鳌山科技创新计划资助项目(2015ASKJ02;2015ASKJ02-03-03)资助;鲆鲽类产业技术体系项目(nycytx-50);中国科学院青促会项目(KFJ-EW-STS-060);国家科技基础条件平台建设运行项目(2016DKA30470)~~

摘　　要：为研究孕激素在大菱鲆(Scophthalmus maximus)精原细胞增殖到减数分裂过程中的作用,以6~22月龄的大菱鲆精巢为材料,通过组织学方法和定量amh、sycp3基因确定精原细胞增殖期和减数分裂期的大菱鲆精巢发育过程。通过酶联免疫吸附技术(ELISA)检测6~22月龄雄性大菱鲆血清中的孕酮(P4)和17α,20β,双羟孕酮(DHP)含量变化,利用q RT-PCR技术和原位杂交技术检测孕酮核受体(pgr)和膜受体(m PRα)在不同组织和不同发育时期的表达情况。结果表明孕激素在精原细胞增殖期高表达,开始出现初级精母细胞时降低,在精子细胞数量增加时表达量再次升高,在精子细胞变态形成精子时降低。pgr在减数分裂初期定位于精巢中的sertoli细胞,在精原细胞增殖至减数分裂期表达量逐渐增加,出现精子后显著降低;m PRα在精原细胞增殖和初级精母细胞增加时表达量都很低,在精子细胞不断增加时显著增加。推测在精原细胞增殖和减数分裂阶段孕激素可能主要通过调节pgr的表达量来促进精巢发育,而在减数分裂II期pgr和m PRα都发挥作用,在精子细胞变态成熟时,主要是m PRα发挥作用。To explore the function of progestin during spermatogonial proliferation and meiosis,we used histological methods to investigate the gonadal development process of the male turbot（Scophthalmus maximus） 6 to 22 months of age.We detected the expression of amh and sycp3 via the real-time quantitative polymerase chain reaction（q RT-PCR）.Using enzyme-linked immunosorbent（ELISA） technology,q RT-PCR,and in-situ hybridization,we detected the serum P4 and DHP levels and the expression patterns of the nuclear progesterone receptor（pgr） and membrane progestin receptor alpha（m PRa） during different development periods.The results reveal that the P4 and DHP levels were high during spermatogonial proliferation and meiosis Ⅱ.The pgr expression was high from spermatogonial proliferation to meiosis I,whereas the mPRα expression was high during meiosis Ⅱ and spermiogenesis.Our in-situ hybridization results indicate pgr m RNA to be predominantly located in the Sertoli cells that were in contact with the proliferating spermatogonia and primary spermatocyte and that the mPRα m RNA was undetectable.Taken together,our data indicate the pgr expression to be involved in mediating the progestin stimulation of spermatogonia proliferation and meiosis,whereas the m PRa plays an important role in meiosis Ⅱ and spermiogenesis.

关 键 词：大菱鲆(Scophthalmus maximus) 减数分裂 孕激素 pgr m PRα 

分 类 号：Q256[生物学—细胞生物学]