半滑舌鳎抗缪勒氏管激素(AMH)基因的克隆及组织表达分析  

Cloning and expression of anti-Müllerian hormone gene in halfsmooth tongue-sole,Cynoglossus semilaevis

作　　者：刘姗姗[1,2] 孙冰[1,3] 梁卓[1,2] 张静[1,2] 陈松林[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,山东青岛266071 [2]上海海洋大学水产与生命学院,上海201306 [3]大连海洋大学辽宁省水生生物学重点实验室,辽宁大连116023

出　　处：《中国水产科学》2013年第1期35-43,共9页Journal of Fishery Sciences of China

基　　金：国家863计划项目(2012AA092203);国家自然科学基金项目(31072202,41006107);山东省泰山学者专项

摘　　要：本研究克隆了半滑舌鳎(Cynoglossus semilaevis)AMH基因并对其进行表达分析。该基因cDNA序列开放阅读框长为1 563 bp,编码蛋白含520个氨基酸。该蛋白含有TGF-β超家族的特征序列,包含1个AMH-N区域和TGF-β结构域,并在C端生物活性区含有9个保守的半胱氨酸残基。系统进化分析表明,半滑舌鳎AMH和所有鱼类AMH聚为一簇,与鲽形目相似性最高。实时定量结果表明,AMH基因在半滑舌鳎不同组织中有差异表达,在正常雄鱼和伪雄鱼性腺中表达量最高。胚后不同时期性腺的实时定量结果表明,性腺分化之前AMH在精巢中相对表达量较低,70 dph(days post hatching)达到最高峰,而在卵巢中呈现先升高后下降的趋势,预示AMH基因可能在半滑舌鳎性腺发育中起重要作用。较正常雄鱼后代而言,AMH基因在伪雄鱼后代的雄鱼和伪雄鱼性腺中的表达都有升高的趋势,而在雌鱼中无明显差异,也预示其可能参与半滑舌鳎的性反转过程。Anti-Müllerian hormone（AMH）,also known as Müllerian inhibiting substance（MIS）,is a glycoprotein belonging to the transforming growth factor β superfamily,which plays a major role during reproductive development in vertebrates.We isolated and characterized AMH cDNA in the half-smooth tongue-sole,Cynoglossus semilaevis.The open reading frame（ORF） of this gene spans a region of 1 563 bp and codes 520 amino acids. Sequence alignment analysis revealed that the AMH of half-smooth tongue-sole and pleuronectiformes shared 61.7% homology.We evaluated the pattern of AMH mRNA expression in 14 tissues of the half-smooth tongue-sole.Expression level was highest in male and neo-male gonads,followed by the blood,skin,and brain. AMH expression was low prior to 70 dph（days post hatching） but was highest in the testis 70 dph.In the ovary, AMH expression increased then decreased,suggesting it plays a role during reproductive development.Furthermore, in contrast with normal offspring,the gonad of male and neo-male non-normal offspring exhibited an increased in expression,but there was no change in females.This suggests that the AMH gene is required for sex reversal.

关 键 词：半滑舌鳎 抗缪勒氏管激素(AMH) 克隆 表达 实时定量 

分 类 号：S917[农业科学—水产科学]