抚仙金线鲃促性腺激素亚基基因克隆及组织表达分析
Gonadotropin Subunits cDNA Cloning and Tissue Expression in Sinocyclocheilus tingi

中文摘要：抚仙金线鲃 Sinocyclocheilus tingi是云南抚仙湖特有种，虽然实现了人工繁殖，但在塘养环境下仍无法自然繁衍。鱼类的生殖活动受下丘脑-垂体-性腺轴的调控，其中促性腺激素在该过程中起重要作用。本实验利用实时荧光定量PCR和cDNA末端快速扩增技术从抚仙金线鲃垂体中克隆了GTHα、FSHβ和LHβ 3种基因的cDNA序列，其中GTHα的开放阅读框（ORF）长度为357 bp，编码118个氨基酸，有10个半胱氨酸残基和2个N-糖基化位点；FSHβ cDNA全长为856 bp，ORF长度为393 bp，编码130个氨基酸，有11个半胱氨酸残基和1个N-糖基化位点；LHβ cDNA全长为930 bp，ORF长度为441 bp，编码146个氨基酸，有12个半胱氨酸残基和1个N-糖基化位点。实时荧光定量PCR结果显示，GTHα和LHβ都只在雄鱼和雌鱼的垂体中表达。FSHβ在垂体中表达量最高，在雌鱼和雄鱼的脂肪、肌肉和雄鱼的精巢和肝脏中有少量表达。本研究为抚仙金线鲃的人工繁育提供了重要的基础数据。
英文摘要： Sinocyclocheilus tingi is an endemic species of the Fuxian Lake in Yunnan province. Although the artificial breeding has been successfully developed, the natural reproduction of S. tingiis still failed in pond culture. It is well known that fish reproduction is regulated by the hypothalamus-pituitary-gonadal axis, and the gonadotropins (GTHs) play important role in this process. In this study, the sequences of GTHα, FSHβ and LHβ were obtained from the pituitary of S. tingi using real-time quantitative PCR (qPCR) and rapid amplification of cDNA ends technology. The results showed that the open reading frame of GTHα was 357 bp, encoding 118 amino acids, containing 10 cysteine knot motifs and 2 N-linked glycosylation sites. The full length of FSHβ cDNA was 856 bp, with 393 bp open reading frame in length encoding the proteins of 130 amino acids, containing 11 cysteine knot motifs and a N-linked glycosylation site. The full length of LHβ cDNA was 930 bp, with 441 bp open reading frame in length encoding proteins of 146 amino acids, containing 12 cysteine knot motifs and a N-linked glycosylation site. The result of qPCR indicated that GTHα and LHβ were only expressed in the pituitary of both male and female fish. The highest expression level of FSHβ was observed in the pituitary, and the even lower levels were detected in the fat tissue, muscle of male and female fish, and testis and liver of male fish. These findings provided basic data for further research in the reproduction of S. tingi. 2016,35(5): 686-690 收稿日期：2015-09-08 DOI：10.11984/j.issn.1000-7083.20160271 分类号：Q959.4;Q78 基金项目：中山大学高校基本科研业务费项目；深圳市战略性新兴产业发展专项资金项目（NYSW20140401010064）；云南省应用基础研究面上项目（2012FB183）；中国科学院昆明动物研究所“一三五”重大专项（Y206B51181）；云南省生物多样性保护专项资金项目 作者简介：杨国坤(1988-),男,博士研究生,研究方向:鱼类内分泌功能基因,E-mail:ygk5210207@126.com *通讯作者：杨君兴,E-mail:yangjx@mail.kiz.ac.cn;李文笙,E-mail:lsslws@mail.sysu.edu.cn 参考文献： 潘晓赋, 刘淑伟, 李再云, 等. 2009. 抚仙金线鲃人工繁殖与鱼苗培育技术[J]. 动物学研究, 30(4): 463-467. 王伟营, 杨君兴, 陈小勇. 2011. 云南境内南盘江水系鱼类种质资源现状及保护对策[J]. 水生态学杂志, 32(5): 19-29. 杨君兴, 陈银瑞. 1995. 抚仙湖鱼类生物学和资源利用[M]. 昆明: 云南科技出版社. 赵亚鹏, 潘晓赋, 杨君兴, 等. 2013. 滇池金线鲃(Sinocyclocheilus