研究报告

斜带石斑鱼(Epinephelus coioides)GnRHⅡ基因的克隆及启动子分析

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  • 中国科学院水生生物研究所, 淡水生态与生物技术国家重点实验室, 武汉 430072

收稿日期: 2012-12-28

  修回日期: 2013-03-21

  网络出版日期: 2013-07-25

基金资助

国家自然科学基金项目(编号:30930069), 中国科学院知识创新重要方向性项目(编号:KSCX2-EW-N-004)和国家重点基础研究发展计划(973计划)项目(编号:2010CB126302)资助

Cloning and promoter analysis of gonadotrophin-releasing hormone Ⅱ in the orange-spotted grouper (Epinephelus coioides)

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  • State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China

Received date: 2012-12-28

  Revised date: 2013-03-21

  Online published: 2013-07-25

摘要

促性腺激素释放激素(Gonadotrophin-releasing hormone, GnRH)是调控脊椎动物生殖的关键因子之一。文章克隆了经济鱼类斜带石斑鱼(Epinephelus coioides)GnRHⅡ基因的cDNA、基因组序列及其启动子, 发现GnRHⅡ 基因启动子中与组织特异性表达相关的区域位于距转录起始点上游-2005 bp~-956 bp之间。以GnRHⅡ启动子驱动绿色荧光蛋白在斑马鱼胚胎中表达的结果显示, GnRHⅡ神经元主要分布在中脑附近和眼部。文章为深入了解GnRHⅡ基因表达的调控机制及GnRHⅡ基因的功能提供了基础信息。

本文引用格式

陈戟 罗余山 朱作言 胡炜 . 斜带石斑鱼(Epinephelus coioides)GnRHⅡ基因的克隆及启动子分析[J]. 遗传, 2013 , 35(7) : 896 -902 . DOI: 10.3724/SP.J.1005.2013.00896

Abstract

Gonadotrophin-releasing hormone (GnRH) is a key regulator of reproduction in all vertebrates. We first cloned the cDNA and genomic DNA sequences coding for GnRHⅡ gene in the orange-spotted grouper (Epinephelis coioides), an economically important marine fish, and then cloned its promoter sequence. The region responsible for the cell-specific expression of GnRHⅡ was located between -2005 bp to -956 bp from the translation start site. GnRHⅡ promoter driven EGFP expression in transgenic zebrafish showed that GnRHⅡ-positive neurons were primarily located in the midbrain and in the eyes. Our results provide an improved understanding of the regulatory mechanism and function of GnRHⅡ of E. coioides.

参考文献

[1] Lee VHY, Lee LTO, Chow BKC. Gonadotropin-releasing hormone: regulation of the GnRH gene. FEBS J, 2008, 275(22): 5458-5478.
[2] Lethimonier C, Madigou T, Muñoz-Cueto JA, Lareyre JJ, Kah O. Evolutionary aspects of GnRHs, GnRH neuronal systems and GnRH receptors in teleost fish. Gen Comp Endocrinol, 2004, 135(1): 1-16.
[3] Li SF, Hu W, Wang YP, Zhu ZY. Cloning and expression analysis in mature individuals of two chicken type-II GnRH (cGnRH-II) genes in common carp (Cyprinus car-pio). Sci China C Life Sci, 2004, 47(4): 349-358.
[4] 黎双飞, 胡炜, 汪亚平, 孙永华, 陈尚萍, 朱作言. 鲤鱼sGnRH基因克隆及其在成熟个体的表达分析. 遗传学报, 2004, 31(10): 1072-1081.
[5] Meyer A, Van de Peer Y. From 2R to 3R: evidence for a fish-specific genome duplication (FSGD). Bioessays, 2005, 27(9): 937-945.
[6] Hu W, Li SF, Tang B, Wang YP, Lin HR, Liu XC, Zou J, Zhu ZY. Antisense for gonadotropin-releasing hormone reduces gonadotropin synthesis and gonadal development in transgenic common carp (Cyprinus carpio). Aquacul-ture, 2007, 271(1-4): 498-506.
[7] Xu J, Huang W, Zhong CR, Luo DJ, Li SF, Zhu ZY, Hu W. Defining global gene expression changes of the hypotha-lamic-pituitary-gonadal axis in female sGnRH-antisense transgenic common carp (Cyprinus carpio). Plos One, 2011, 6(6): e21057.
[8] Zandbergen MA, Kah O, Bogerd J, Peute J, Goos HJ. Expression and distribution of two gonadotropin-releasing hormones in the catfish brain. Neuroendocrinol, 1995, 62(6): 571-578.
[9] Gothilf Y, Muñoz-Cueto JA, Sagrillo CA, Selmanoff M, Chen TT, Kah O, Elizur A, Zohar Y. Three forms of gonadotropin-releasing hormone in a perciform fish (Sparus aurata): complementary deoxyribonucleic acid characterization and brain localization. Biol Reprod, 1996, 55(3): 636-645.
[10] González-Martínez D, Zmora N, Mananos E, Saligaut D, Zanuy S, Zohar Y, Elizur A, Kah O, Muñoz-Cueto JA. Immunohistochemical localization of three different prepro-GnRHs in the brain and pituitary of the European sea bass (Dicentrarchus labrax) using antibodies to the corresponding GnRH-associated peptides. J Comp Neurol, 2002, 446(2): 95-113.
[11] Palevitch O, Kight K, Abraham E, Wray S, Zohar Y, Gothilf Y. Ontogeny of the GnRH systems in zebrafish brain: in situ hybridization and promoter-reporter expression analyses in intact animals. Cell Tissue Res, 2007, 327(2): 313-322.
[12] Abraham E, Palevitch O, Ijiri S, Du SJ, Gothilf Y, Zohar Y. Early development of forebrain gonadotrophin-releasing hormone (GnRH) neurones and the role of GnRH as an autocrine migration factor. J Neuroendocrinol, 2008, 20(3): 394-405.
[13] Hu SY, Chen MH, Lin YC, Lin GH, Gong HY, Yang TH, Wu JL. Cloning and functional analysis of the proximal promoter region of the three GnRH genes from the silver sea bream (Sparus sarba). Comp Biochem Physiol B Biochem Mol Biol, 2008, 151(4): 373-380.
[14] Kah O, Lethimonier C, Somoza G, Guilgur LG, Vaillant C, Lareyre JJ. GnRH and GnRH receptors in metazoa: a historical, comparative, and evolutive perspective. Gen Comp Endocrinol, 2007, 153(1-3): 346-364.
[15] Kitahashi T, Sato H, Sakuma Y, Parhar IS. Cloning and functional analysis of promoters of three GnRH genes in a cichlid. Biochem Biophy Res Commun, 2005, 336(2): 536-543.
[16] Wang Y, Zhou L, Yao B, Li CJ, Gui JF. Differential expression of thyroid-stimulating hormone β subunit in gonads during sex reversal of orange-spotted and red-spotted groupers. Mol Cell Endocrinol, 2004, 220(1-2): 77-88.
[17] Chen J, Hu W, Zhu ZY. Progress in studies of fish repro-ductive development regulation. Chin Sci Bull, 2013, 58(1): 7-16.
[18] Shi Y, Zhang Y, Li SS, Liu Q, Lu DQ, Liu M, Meng ZN, Cheng CHK, Liu XC, Lin HR. Molecular identification of the kiss2/kiss1ra system and its potential function during 17alpha-methyltestosterone-induced sex reversal in the orange-spotted grouper, Epinephelus coioides. Biol Reprod, 2010, 83(1): 63-74.
[19] Zhang Y, Li SS, Liu Y, Lu DQ, Chen HP, Hua
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