Expression Characteristics Analysis of Gonadal Aromatase Gene cyp19a1a in Tissues and During Gonad Development Stage of Hemibagrus guttatus
LI Guang-Lue1,2, CHEN Yan-Yun1, FANG Jun-Chao2, HU Dan2, LUO Wen-Yin2, ZHU Jia-Jie1,*, HU Qiao-Mu2,*
1 Guangxi Key Laboratory of Polysaccharide Materials and Modification, Guangxi Minzu University, Nanning 530006, China; 2 Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
摘要斑鳠(Hemibagrus guttatus)是珍贵的淡水鱼类,性别差异是其生殖繁育的基础,但目前斑鳠性别分化机制尚不清楚。细胞色素P450家族19亚家族A成员1A(cytochrome P450 family 19 subfamily A member 1A, cyp19a1a)基因是影响雌激素生物合成的关键基因,对鱼类性别分化和卵巢发育至关重要。为探究cyp19a1a基因在斑鳠性别分化中的作用机制,本研究采用qPCR技术分析斑鳠不同组织、不同发育时期和性逆转个体性腺中cyp19a1a表达量的变化;通过荧光原位杂交技术(fluorescence in situ hybridization, FISH)确定cyp19a1a在斑鳠性腺细胞中的表达。结果显示,斑鳠cyp19a1a ORF序列长1 587 bp,编码528个氨基酸,包含芳香化酶氨基酸序列的保守功能区。qPCR结果表明,cyp19a1a在斑鳠各个部位均有表达,在性腺中的表达量最高,其次为垂体和肝脏;伪雌鱼cyp19a1a的表达特征与同龄正常雌鱼一致,而且卵巢中该基因的表达量显著高于同期精巢(P<0.05),并且随性腺发育逐渐升高。FISH结果显示,cyp19a1a在精子和成熟卵母细胞中的信号较弱,在卵原细胞和早期卵母细胞中信号较强。以上结果表明,cyp19a1a基因在斑鳠性腺发育和生殖细胞生长发育及性逆转过程中发挥重要作用。本研究为解析斑鳠cyp19a1a基因的功能及其性别分化机制的研究提供参考。
Abstract：Hemibagrus guttatus is a rare freshwater fish species, sexual dimorphism is the basis of its reproduction, but the mechanism of sex differentiation in H. guttatus remains unclear. The cytochrome P450 family 19 subfamily A member 1A(cyp19a1a) is a critical gene that influences estrogen biosynthesis and is essential for both sex differentiation and ovarian development. To investigate the mechanism of the cyp19a1a gene in sex differentiation of H. guttatus, qPCR was used to analyze the expression levels of cyp19a1a in different tissues, at different developmental stages, and in the gonads of sex-reversed individuals. Fluorescence in situ hybridization (FISH) was employed to determine the expression of cyp19a1a in gonadal cells of H. guttatus. The results revealed that the ORF sequence of cyp19a1a in H. guttatus was 1 587 bp in length, encoding 528 amino acids, and contained conserved functional domains of the aromatase amino acid sequence. The qPCR results showed that cyp19a1a was expressed in all examined tissues of H. guttatus, with the highest expression levels in the gonads, followed by the brain and liver. The expression profile of cyp19a1a in pseudofemales was consistent with that of normal females at the same age. Moreover, the expression of cyp19a1a in ovaries was significantly higher than that in testes at corresponding stages (P<0.05) and gradually increased with gonadal development. FISH results showed that the signal for cyp19a1a was weaker in sperm and mature oocytes but stronger in oogonia and early oocytes. The above findings indicated that the cyp19a1a gene played an important role in gonadal development, germ cell growth, and sex reversal in H. guttatus. This study provides a reference for understanding the function of the cyp19a1a gene and the sex determination mechanisms in H. guttatus.
李光略, 陈燕芸, 方俊超, 胡丹, 罗雯尹, 朱佳杰, 胡乔木. 性腺型芳香化酶cyp19a1a基因在斑鳠组织及性腺发育时期的表达特征分析[J]. 农业生物技术学报, 2026, 34(7): 1517-1529.
LI Guang-Lue, CHEN Yan-Yun, FANG Jun-Chao, HU Dan, LUO Wen-Yin, ZHU Jia-Jie, HU Qiao-Mu. Expression Characteristics Analysis of Gonadal Aromatase Gene cyp19a1a in Tissues and During Gonad Development Stage of Hemibagrus guttatus. 农业生物技术学报, 2026, 34(7): 1517-1529.
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