红鳍东方鲀幼鱼中性别分化相关基因的表达分析  

Expression Analysis of Genes Related to Sex Differentiation in Tiger PufferTakifugu rubripes Juveniles

作　　者：姜洁明 沈旭芳 刘鹰[1,2,4] 张琦 周慧婷[1] 王佳 闫红伟[1,2] 王辰奇[1] 刘奇 JIANG Jieming;SHEN Xufang;LIU Ying;ZHANG Qi;ZHOU Huiting;WANG Jia;YAN Hongwei;WANG Chenqi;LIU Qi(Dalian Ocean University, Dalian 116023, China;Key Laboratory of Environment Controlled Aquaculture, Ministry of Education, Dalian 116023, China;Liaoning Normal University, Dalian 116029, China;Pilot National Laboratory for Marine Science and Technology, Qingdao 266237, China)

机构地区：[1]大连海洋大学,辽宁大连116023 [2]设施渔业教育部重点实验室,辽宁大连116023 [3]辽宁师范大学,辽宁大连116029 [4]青岛海洋科学与技术试点国家实验室,山东青岛266237

出　　处：《水产科学》2022年第3期362-369,共8页Fisheries Science

基　　金：国家重点研发计划项目(2019YFD0900503);辽宁省重点研发计划项目(2019JH2/10200015);大连市科技创新基金项目(2018J12SN069);大连市青年科技之星项目(2019RQ130);大连海洋大学大学生创新创业训练计划项目(B202010158004).

摘　　要：为查明性别分化相关基因在红鳍东方鲀性别分化关键时期的表达规律,以孵化后40、60、80 d的红鳍东方鲀幼鱼为试验对象,对性腺进行组织学观察,并对采样个体进行遗传性别鉴定,采用实时荧光定量PCR(qRT-PCR)的方法研究amhr2、dmrt1、gsdf、cyp19a1a、foxl2、dnmt1基因在遗传型为XX(雌性)和XY(雄性)幼鱼性腺中的表达规律。结果显示,上述基因的表达呈现出明显的性别二态性,其中amhr2基因在孵化后40、60 d XX和XY个体性腺中的表达量差异不显著(P>0.05),但在孵化后80 d的XX个体性腺中的表达量显著高于在XY个体性腺中的表达量(P<0.05)。Dmrt1和gsdf基因从孵化后20 d开始在XY个体性腺中的表达量显著上升(P<0.05),且在所有采样时间点的表达量均显著高于在XX个体性腺中的表达量(P<0.05)。Cyp19a1a和foxl2基因在孵化后40、60、80 d的XX个体性腺中的表达量显著高于在XY个体性腺中的表达量(P<0.05)。而dnmt1基因在孵化后40、60 d的XX和XY个体性腺中的表达量差异不显著(P>0.05),但在孵化后80 d时XY个体性腺中的表达量显著高于XX个体性腺中的表达量(P<0.05)。研究结果表明,dmrt1和gsdf基因可能在红鳍东方鲀精巢分化过程中具有重要的作用,cyp19a1a和foxl2可能在卵巢分化过程中具有重要的作用。以上试验结果可为深入研究性别分化相关基因在红鳍东方鲀性别分化过程中的功能奠定基础。In order to clarify the expression patterns of sex differentiation-related genes during the critical period of sex differentiation in tiger puffer Takifugu rubripes,the histological characteristics of the gonads were observed in tiger puffer juveniles at 40,60 and 80 d post hatching firstly.Then,after the genetic sex verification of individual,the expression patterns of amhr2,dmrt1,gsdf,cyp19a1a,foxl2 and dnmt1 genes were determined in the gonads of XX and XY tiger puffer juveniles by real-time quantitative PCR(qPCR).The results showed that the expression of the above genes showed sexual dimorphism obviously,without significant difference in the expression level of amhr2 gene in the gonads of XX and XY tiger puffer juveniles at 40 and 60 d post hatching(P>0.05),significantly higher expression level of amhr2 gene in the gonads of XX juveniles than that in XY juveniles at 80 d post hatching(P<0.05).The expression levels of dmrt1 and gsdf genes in the gonads of XY juveniles were increased significantly from 20 d,and the expression levels in the gonads of XY juveniles were significantly higher than that in XX juveniles at all the three sampling points(P<0.05).The expression levels of cyp19a1a and foxl2 genes in the gonads of XX juveniles were significantly higher than that in XY juveniles at the 40,60 and 80 d post hatching(P<0.05).There was no significant difference in the expression level of dnmt1 gene in the gonads of XX and XY tiger puffer juveniles at 40 and 60 d post hatching(P>0.05),but the expression level of dnmt1 gene in the gonads of XY tiger puffer juveniles was significantly higher than that in XX juveniles at 80 d post hatching(P<0.05).The findings demonstrated that dmrt1 and gsdf genes played critical roles in testicular differentiation,while cyp19a1a and foxl2 genes played critical roles in ovarian differentiation in tiger puffer juveniles.The findings laid a foundation for functional analysis of the sex differentiation-related genes during the development of sex differentiation of tiger puffer.

关 键 词：红鳍东方鲀 幼鱼 性别分化相关基因 基因表达 

分 类 号：Q786[生物学—分子生物学]