| 摘要: |
| 密斑刺鲀(Diodon hystrix)是南海名特优鱼类, 同时具备食用和中药保健价值, 但目前关于密斑刺鲀生殖生理的研究较少, 影响了该鱼繁育理论研究的发展。众所周知, 促性腺激素(Gonadotropic hormone, GtH)是生殖调控的关键激素之一, 在生殖调控轴中发挥着承上启下的作用。利用转录组分析及分子克隆技术从密斑刺鲀中克隆并鉴定出fshβ和lhβ基因的cDNA序列, 通过生物信息学分析, 发现密斑刺鲀fshβ cDNA开放阅读框共363 bp, 编码120个氨基酸; lhβ cDNA开放阅读框共420 bp, 编码139个氨基酸。通过序列比对和系统进化树分析, 表明密斑刺鲀Fshβ和Lhβ均与鲀科类的鱼的亲缘关系较近。利用三级结构分析, 发现Fshβ的三级结构相对保守, 而Lhβ的三级结构差异较大。利用实时荧光定量的方法检测了密斑刺鲀fshβ和lhβ基因的组织分布情况, 结果表明密斑刺鲀fshβ和lhβ在垂体的表达量最高, 其次是性腺。此外, 进一步检测了fshβ和lhβ基因在雌雄性腺发育过程中的表达模式, 发现垂体中fshβ基因伴随性腺的发育, 表达量逐渐下降, 而lhβ基因的表达量呈逐渐增加模式。研究结果将为构建密斑刺鲀生殖调控网络提供研究基础。 |
| 关键词: 密斑刺鲀 fshβ lhβ 分子克隆 表达模式 |
| DOI：10.11693/hyhz20230400098 |
| 分类号:S965.3 |
| 基金项目:海南省重点研发计划,ZDYF2018225号;湛江市科技计划项目,2022A01214号;阳江市科技计划项目,2022011号,SDZX2023027号 |
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| CLONING AND EXPRESSION ANALYSIS OF fshβ AND lhβ GENE IN DIODON HYSTRIX |
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GUO Yu-Wen, LI Yi-Kai, DAI Ming-Shu, LIANG Jun-Hao, HUANG Yan-Lin, LI Guang-Li, CHEN Hua-Pu
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Guangdong Research Center on Reproductive Control and Breeding Technology of Indigenous Valuable Fish Species, Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding, Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China
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| Abstract: |
| Diodon hystrix is a well-known economic fish in the South China Sea with high edible and medicinal value. However, studies on the reproduction of D. hystrix are few, which hinder the development of its artificial breeding. Gonadotropic hormone (GtH) is one of the key hormones for reproductive regulation, and plays a connecting role in the reproductive regulation axis. We cloned the cDNA sequences of fshβ and lhβ genes from D. hystrix by molecular cloning techniques. Through bioinformatics analysis, we found that the open reading frame (ORF) of fshβ cDNA contains 363 bp, encoding 120 amino acids, while the ORF of lhβ cDNA contains 420 bp, encoding 139 amino acids. Sequence alignment and phylogenetic tree analyses showed that both Fshβ and Lhβ of D. hystrix are similar to those of fishes of the family Tetraodontidae. According to the tertiary structure analysis, we found that the tertiary structure of Fshβ is highly conservative, while the tertiary structure of Lhβ is moderately conservative. The tissue distribution of fshβ and lhβ genes in D. hystrix was detected by qRT-PCR. Results show that the expression of both fshβ and lhβ was the highest in the pituitary, followed by the gonads. In addition, we detected the expression patterns of fshβ and lhβ genes during the development of male and female gonads, and found that the expression of fshβ gene in the pituitary gradually decreased with the development of gonads, while the expression of lhβ gene gradually increased. This study provided a theoretic basis for constructing the reproductive regulatory network of D. hystrix. |
| Key words: Diodon hystrix fshβ lhβ molecular cloning expression pattern |
