过表达cyp19a对青鳉(Oryzias latipes)性腺发育的影响  

Effect of Overexpression of cyp19a on Gonadal Development in Medaka(Oryzias latipes)

作　　者：王梦洋 黄岩[1,2,3] 陈磊 戴荣贵 黄名锡[1,2,3] 蒋月雯 陈天圣 WANG Mengyang;HUANC Yan;CHEN Lei;DAI Ronggui;HUANG Mingxi;JIANG Yuewen;CHEN Tiansheng(State Key Laboratory of Mariculture Breeding,College of Fisheries,Jimei University,Xiamen,361021;Engineering Research Center of the Modern Technology for Eel Industry,Ministry of Education,Xiamen,361021;Key Laboratory of Healthy Mariculture for the East China Sea,Ministry of Agriculture and Rural Affairs,Xiamen,361021)

机构地区：[1]集美大学水产学院,海水养殖生物育种全国重点实验室,厦门361021 [2]鳗鲡现代产业技术教育部工程研究中心,厦门361021 [3]农业农村部东海海水健康养殖重点实验室,厦门361021

出　　处：《基因组学与应用生物学》2025年第1期53-63,共11页Genomics and Applied Biology

基　　金：国家自然科学基金项目(32273127);集美大学科研基金项目(ZQ2020003)共同资助。

摘　　要：cyp19a基因编码的芳香化酶是一个重要的类固醇合成酶,能够促进雄激素向雌激素转化,影响动物卵巢的正常生理功能。在许多鱼类中,敲除cyp19a基因会导致雌鱼性逆转为雄鱼,但过表达cyp19a基因能否促进雄鱼逆转为雌鱼仍不清楚。本研究通过构建cyp19a转基因青鳉(Oryzias latipes)Tg(EFa1:olcyp19a),从DNA、RNA、蛋白水平的表达及组织形态学等层面探讨过表达cyp19a基因对青鳉性腺发育的影响。本研究构建了pBOS-cyp19a真核表达载体并将其显微注射入青鳉胚胎,通过PCR筛选对插入拷贝数和蛋白水平进行分析,成功获得了两个具有不同拷贝数的cyp19a转基因品系。RT-qPCR结果显示,转基因雌鱼卵巢中cyp19a mRNA的表达量显著增加,雄鱼精巢中的表达量也显著升高,超过了野生型雌鱼卵巢中的水平。然而,转基因雄鱼精巢中与精巢发育相关的基因dmrt1、gsdf、tex9、myb和sycp2的表达无显著变化。此外,转基因的XY雄鱼未出现性逆转,其繁殖能力与对照组相比无显著差异。综上所述,单独过表达cyp19a不足以促进青鳉发生由雄性向雌性的性逆转,表明cyp19a是雌性卵巢发育的必要因素,但并非决定性因素。The cyp19a gene encodes aromatase,which is an important steroid synthase that promotes the conversion of androgens to estrogens,affecting the normal physiological function of animal ovaries.In many fish species,knocking out the cyp19a gene can lead to sex reversal from females into males,but it remains unclear whether overexpression of the cyp19a gene can induce the reversal of males to females.This study constructed transgenic medaka fish(Oryzias latipes)Tg(EFa1:olcyp19a)to explore the effects of cyp19a gene overexpression on gonadal development at multiple levels,including DNA,RNA,protein level expression,and tissue morphology.This study successfully constructed the pBOS-cyp19a eukaryotic expression vector,and performed microinjections of it into the embryos of medaka.The insertion copy number and protein level were analyzed through PCR screening,and two transgenic lines with different copy numbers of cyp19a were successfully obtained.RT-qPCR results showed that the expression level of cyp19a mRNA was significantly increased in the ovaries of transgenic female fish,and the expression level in the testes of transgenic males was also significantly elevated,exceeding that of wild-type ovaries.However,there were no significant differences in the expression of testis development-related genes dmrt1,gsdf,tex9,myb,and sycp2 in the testes of transgenic males.Furthermore,XY transgenic males did not exhibit sex reversal,and their reproductive capacity did not differ significantly from that of the control group.In summary,the overexpression of cyp19a alone does not promote sex reversal from male to female in medaka,indicating that cyp19a is necessary but not a decisive factor for ovarian development in females.

关 键 词：青鳉(Oryzias latipes) cyp19a 转基因 性腺发育 

分 类 号：S917.4[农业科学—水产科学]