大菱鲆figla基因的克隆及其在卵巢分化过程中的表达分析
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1.海水养殖生物育种与可持续产出全国重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛 266071 ;2.上海海洋大学水产与生命学院 上海 201306 ;3.浙江海洋大学水产学院 浙江 舟山 316022 ;4.青岛通用水产养殖有限公司 山东 青岛 266404 ;5.烟台开发区天源水产有限公司 山东 烟台 264001

作者简介:

孙晓璇,E-mail:xiyi7941@foxmail.com

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中图分类号:

S917.4

基金项目:

国家科技创新 2030 (2023ZD0405505)、国家重点研发计划 (2022YFD2400402)、国家海水鱼产业技术体系 (CARS-047-G31)和山东省重点研发计划(2019GHY112023)共同资助


Cloning and Expression Analysis of figla During Ovarian Differentiation in Turbot (Scophthalmus maximus)
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1.State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences, Qingdao 266071 , China ;2.College of Fisheries and Life Sciences, Shanghai Ocean University, Shanghai 201306 , China ;3.College of Fisheries, Zhejiang Ocean University, Zhoushan 316022 , China ;4.Qingdao General Aquaculture Co., Ltd., Qingdao 266404 , China ;5.Yantai Development Zone Tianyuan Aquatic Products Co., Ltd., Yantai 264001 , China

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    摘要:

    figla (factor in the germ cell line, alpha)是生殖细胞中特异性表达的转录因子之一,对于调控鱼类原始卵泡形成和卵巢发育起着关键作用。为了明确大菱鲆(Scophthalmus maximus) figla 基因序列及其在卵巢早期发育过程中的功能,本研究以大菱鲆全雌苗种为实验对象,采用 RT-PCR、cDNA 末端快速扩增(RACE)克隆获得大菱鲆 figla 基因的全长 cDNA 序列,并通过半定量 PCR、实时荧光定量 PCR (qRT-PCR)、原位杂交(ISH)对其组织表达分布和卵巢早期发育过程中的时空表达模式进行了研究。结果显示,大菱鲆 figla 基因 cDNA 全长为 1 006 bp,开放阅读框为 609 bp (150~758 bp), 编码 202 个氨基酸,具有碱性螺旋–环–螺旋(basic Helix-Loop-Helix, bHLH)序列特征;figla 在大菱鲆性腺中特异表达,卵巢中的表达水平高于精巢;在不同发育时期的卵巢发育过程中,figla mRNA 于孵化后 25 日龄(dph)性腺中开始表达,35 dph 显著增加,此后直至 90 dph 表达水平持续增加;原位杂交结果显示,figla mRNA 主要定位于卵母细胞胞质内。研究结果初步提示,figla 基因与大菱鲆卵巢和卵母细胞分化密切相关,在促进卵泡形成和性腺分化方面起到关键作用。本研究结果为深入探索大菱鲆卵巢分化和发育的分子调控机制提供了参考依据。

    Abstract:

    The transcription factor gene figla (factor in the germ line, alpha), a member of the basic helix-loop-helix (bHLH) family, has been extensively documented for its pivotal role in mammalian ovarian development and primordial follicle formation. However, studies of figla in teleosts are less detailed than those in mammals. Turbot (Scophthalmus maximus), an important aquaculture species in Europe and China, exhibits sexual dimorphism in growth and body size and possesses a female heterogametic sex determination system (ZW female and ZZ male). Therefore, it is crucial to understand the mechanisms underlying sex determination and differentiation. This study exclusively used female individuals as experimental subjects to elucidate the complete cDNA sequence of figla in turbot and its role during early ovarian development. The full-length cDNA sequence of figla was cloned using RT-PCR and rapid amplification of cDNA ends techniques. Tissue expression distribution and spatiotemporal expression patterns during early ovarian development were investigated using semi-quantitative RT-PCR, real-time fluorescence quantitative PCR, and in situ hybridization. The full length of the figla gene cDNA had 1,006 base pairs (bp), with an open reading frame ranging from 150 bp to 758 bp, encoding 202 amino acids with a conserved bHLH domain. In the turbot gonads, figla exhibited specific expression, showing higher levels in the ovaries than in the testes. The expression pattern of figla mRNA was detected throughout various stages of ovarian development, commencing at 25 dph and progressively increasing until reaching its peak at 90 dph. In situ hybridization experiments revealed the predominant localization of figla mRNA within the cytoplasmic region of oocytes. In conclusion, the distinct expression pattern of figla in male and female turbot, its localization within primary oocytes, and its expression patterns during early ovarian development suggest a crucial role for this gene in the normal development of oocytes and ovaries. Further research is required to investigate the regulatory mechanisms underlying oocyte development and gonadal differentiation. The study of figla function not only enhances our understanding of sex determination and differentiation mechanisms in turbot but also provides novel insights and methodologies for sex control and breeding applications in this species.

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孙晓璇, 成路遥, 孟振, 徐文腾, 刘新富, 张和森, 徐荣静. 大菱鲆 figla 基因的克隆及其在卵巢分化过程中的表达分析. 渔业科学进展, 2025, 46(6): 167–176

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  • 收稿日期:2024-11-28
  • 最后修改日期:2025-02-08
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  • 在线发布日期: 2025-11-10
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