| 本文已被：浏览 730次 下载 97次 |
码上扫一扫！ |
| 暗纹东方鲀伪雄鱼制备及其性逆转相关基因FOXH1 的克隆和组织表达分析 |
|
臧雪纯1,2, 唐晓东1,2, 刘雨曦1,2, 赵诚1,2, 谢骏1,2, 刁统治1,2, 褚鹏1,2, 尹绍武1,2, 王涛1,2
|
|
1.南京师范大学 海洋科学与工程学院，江苏 南京 210023;2.江苏省特色水产育种与绿色高效养殖技术工程研究中心，江苏 南京 210023
|
|
| 摘要: |
| 【 目的】使用外源雄激素17α-甲基睾酮（17α-MT）诱导性别发育关键期5~150日龄的暗纹东方鲀，制 备伪雄鱼，并探究其性逆转相关基因FOXH1的组织表达特征。【 方法】设置无激素处理的对照组（雌鱼，雄鱼）和30，60 mg/kg 17α-MT处理的试验组（伪雄鱼）。采用性别特异分子标记鉴定试验鱼遗传性别，通过性腺组织学判定其生 理性别。统计增重率、特定生长率和性腺成熟系数，测定性成熟个体精子活力。利用RACE技术克隆暗纹东方鲀性逆 转相关基因FOXH1全长，对其在不同幼鱼性腺和成鱼性腺、脑、肝脏、鳃、心脏和肾脏中的表达水平进行检测。【 结 果】30~150日龄间，试验组鱼增重率和特定生长率显著低于对照组，可见17α-MT抑制了鱼体生长。90和150日龄时， 试验组鱼性腺成熟系数显著低于对照组，表明试验组鱼性腺发育受到抑制；试验组鱼体性腺均呈精巢形态，可观察到精 原细胞和初级精母细胞。2龄和3龄伪雄鱼精子活力均大于对照组（雄鱼），MT30和MT60组伪雄鱼精子活力无显著差 异。暗纹东方鲀FOXH1基因全长2 823 bp，包含开放阅读框1 533 bp，5′ UTR 519 bp，3′ UTR 771 bp，编码510个氨基 酸。暗纹东方鲀FOXH1氨基酸序列与红鳍东方鲀同源性最高（99.41%）。进化分析表明，暗纹东方鲀FOXH1氨基酸 序列与红鳍东方鲀进化关系最近，与菊黄东方鲀、金钱鱼进化关系较近。FOXH1在暗纹东方鲀7种组织（精巢、卵巢、 脑、肝脏、鳃、心脏和肾脏）中均有表达，在卵巢中的表达量显著高于其他组织，伪雄鱼性腺中FOXH1表达显著低于对照 组（P<0.05）。【 结论】17α-MT能够诱导暗纹东方鲀雄性化，使其性腺精巢化发育并具有生理功能，但幼鱼生长受到一 定的抑制；FOXH1基因mRNA在暗纹东方鲀伪雄鱼性腺中表达水平显著降低（P<0.05），推测FOXH1在暗纹东方鲀 性逆转方面发挥着重要作用。 |
| 关键词: 暗纹东方鲀 17α-甲基睾酮 伪雄鱼 性腺发育 FOXH1基因 |
| DOI：10. 13207/j. jnwafu. 2026. 07. 001 |
| 分类号: |
| 基金项目:江苏省农业科技自主创新资金项目（CX（22）2029）；江苏省种业振兴“揭榜挂帅”项目（JBGS［ 2021］ 034）；江苏省第六期“333人才”培养支持项目 |
|
| Induction of pseudo⁃males and cloning and tissue expression analysis of sex reversal related gene FOXH1 in pufferfish （Takifugu fasciatus） |
|
ZANG Xuechun1,2, TANG Xiaodong1,2, LIU Yuxi1,2, ZHAO Cheng1,2, XIE Jun1,2, DIAO Tongzhi1,2, CHU Peng1,2, YIN Shaowu1,2, WANG Tao1,2
|
|
1.College of Marine Science and Engineering，Nanjing Normal University，Nanjing，Jiangsu 210023，China;2.Engineering Research Center for Aquatic Animals Breeding and Green Efficient Aquacultural Technology，Nanjing，Jiangsu 210023，China
|
| Abstract: |
| 【Objective】This study aimed to induce sex differentiation during the critical developmental period（5?150 days of age） of Takifugu fasciatus using exogenous androgen 17α-methyltestosterone（17α-MT） to produce pseudo-male fish，and investigate the expression of the sex-reversal-related gene FOXH1.【Method】A control group（females and males without hormone treatment） and experimental groups treated with 17α-MT at 30 and 60 mg/kg（pseudo-males） were established. The genetic sex of experimental fish was identified using sex-specific molecular markers，and their physiological sex was determined by gonadal histology. Weight gain rate，specific growth rate，and gonadosomatic index（GSI） were calculated，and sperm motility was measured in sexually mature individuals. The full-length FOXH1 gene of T. fasciatus was cloned using RACE technology，and its expression levels were examined in the gonad of juvenile fish and the gonad，brain，liver，gill，heart and kidney of adult fish.【Result】From 30 to 150 days of age，the experimental groups exhibited significantly lower weight gain and specific growth rates compared with the control，indicating growth inhibition by 17α-MT. At 90 and 150 days of age，GSIs in the experimental groups were significantly lower than those of the control，suggesting suppressed gonadal development. Histological analysis showed that gonads of the treated fish developed into testes with observable spermatogonia and primary spermatocytes. In two-and three-year-old pseudo-males，sperm motility was higher than that of control males，with no significant difference between MT30 and MT60 groups. The full-length FOXH1 cDNA of T. fasciatus was 2 823 bp，including a 1 533 bp open reading frame，a 519 bp 5′UTR，and a 771 bp 3′UTR，encoding 510 amino acids. The FOXH1 amino acid sequence of T. fasciatus shared the highest homology with T. rubripes（99.41%）. Phylogenetic analysis revealed that the FOXH1 was most closely related to that of T. rubripes，and showed closer relationships to T.flavidus and Monodactylus argenteus. FOXH1 was expressed in all seven tested tissues（testis，ovary，brain，liver，gill，heart，and kidney），with significantly higher expression in the ovary. Its expression in the gonads of pseudo-males was significantly lower than that in the control group（P<0. 05）.【Conclusion】17α-MT can induce masculinization in T. fasciatus，leading to testicular development with functional spermatogenesis，although juvenile growth is suppressed.The significant downregulation of FOXH1 mRNA in the gonads of pseudo-males（P<0. 05） suggests that FOXH1 plays an important role in sex reversal in T. fasciatus. |
| Key words: Takifugu fasciatus 17α-Methyltestosterone pseudo-male gonadogenesis FOXH1 gene |
