抚仙金线鲃kisspeptin基因的克隆及组织表达特性
Molecular Cloning and Tissue Distribution of Kisspeptin from Sinocyclocheilus tingi

中文摘要：抚仙金线鲃 Sinocyclocheilus tingi是云南抚仙湖的特有种，虽已突破其人工繁殖技术，但在塘养环境下不能自然繁衍。 kiss1基因编码的神经多肽kisspeptin被认为是下丘脑-垂体-性腺轴的重要调节因子，通过调控促性腺激素释放激素、促性腺激素等激素的分泌参与到生殖调控中。本研究采用反转录PCR和cDNA末端快速扩增（RACE）技术在抚仙金线鲃中克隆出编码kisspeptin的2个基因 kiss1和 kiss2的cDNA，其中 kiss1的开放阅读框长度为351 bp，编码116个氨基酸； kiss2的开放阅读框长度为369 bp，编码122个氨基酸。反转录PCR结果显示， kiss1在抚仙金线鲃雌雄个体中均是肠道的表达量最高； kiss2表达量最高的组织在雄性和雌性中不同，雄性为肠道，雌性则是下丘脑和全脑。为进一步探究 kiss1和 kiss2在中枢神经系统及垂体中的分布，本研究中将抚仙金线鲃的脑部分为8个部分进行反转录PCR，结果显示，雄性脊髓和垂体中的 kiss1表达量较高，雌性垂体中的 kiss1表达量最高，其次是视顶盖； kiss2在雌性和雄性中均在下丘脑的表达量最高。本研究的结果为抚仙金线鲃人工繁殖提供了更多探索的方向。
英文摘要： Sinocyclocheilus tingi is an endemic species of Fuxian Lake in Yunnan province. Even though its artificial reproduction is available right now, they can't reproduce in fishponds. Kisspeptin encoded by kiss1 was considered as an important regulator of the hypothalamic-pituitary-gonadal axis and took part in reproduction by controlling the releasing of gonadotropin-releasing hormone, gonadotropin, and etc. In the present research we cloned the cDNA of kiss1 and kiss2 of S. tingi with reverse transcription PCR and rapid amplification of cDNA ends (RACE). The open reading frame of kiss1 was 351 bp and encoded a polypeptide of 116 amino acid and that of kiss2 was 369 bp and encoded a 122-amino-acid polypeptide. Reverse transcription PCR analysis showed that the mRNA expression quantity of kiss1 was the highest in intestine. Likely, the mRNA expression of kiss2 was the highest in intestine in male and hypothalamus and brain in female. To explore the distribution of kiss1 and kiss2 in brain, the brain was divided into eight parts for reverse transcription PCR analysis. The result showed that the highest expression of kiss1 in male was observed in both the spinal cord and the pituitary, which in female was only in the pituitary and followed by the optic tectum. Meanwhile, the kiss2 mRNA got its highest expression in the hypothalamus. 2016, 35(4): 496-502 收稿日期：2015-09-08 DOI：10.11984/j.issn.1000-7083.20150273 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：廖宗甄(1993-),硕士研究生,研究方向:水生生物学,E-mail:237688797@qq.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. Akazome Y, Kanda S, Okubo K, et al. 2010. Functional and evolutionary insights into vertebrate kisspeptin systems from studies of fish brain[J].