双棘黄姑鱼IGF3基因克隆及表达模式分析

IGF3 gene cloning and expression pattern of Protonibea diacanthus

  • 摘要: 以双棘黄姑鱼(Protonibea diacanthus)为实验对象,通过RACE方法克隆得到IGFs家族成员IGF3。双棘黄姑鱼IGF3 cDNA全长1 184 bp。氨基酸一致性分析显示,双棘黄姑鱼IGF3与尼罗罗非鱼(Oreochromis niloticus)一致性最高,仅为58.0%;与自身IGF1的2个亚型及IGF2相比,一致性分别低至26.2%、25.2%和21.9%。系统进化树显示鱼类IGF3基因先与IGF2聚类,再与IGF1汇聚构成IGF家族,最后IGFs与胰岛素聚类。组织表达模式显示IGF3在双棘黄姑鱼心脏、性腺、脑区均有表达,在性腺中表达最强。实时定量PCR结果表明,IGF3在精巢发育期表达量显著高于其他时期,呈显著下降趋势;在卵巢成熟期的表达量显著高于其他时期。此外,雌鱼在成熟期前、中、后脑IGF3的表达量显著高于其他时期,而雄鱼各脑区IGF3表达趋势呈多样性。上述结果表明,IGF3可能参与了双棘黄姑鱼精巢发育和卵巢成熟期生殖功能的调节,且前脑可能主要参与了IGF3在性腺表达的反馈调节。

     

    Abstract: We cloned IGF-Ⅲ gene from Protonibea diacanthus by RACE method. The length of cDNA was 1 184 bp. The homologous analysis of deduced protein shows that the identity of IGF-Ⅲ gene and Oreochromis niloticus was 58.0%. Compared with IGF-Ⅱ and two subtypes of IGF-Ⅰof P.diacantus, the identities were 21.9%, 26.2% and 25.2%, respectively. Phylogenetic tree analysis shows that IGF-Ⅲ tended to cluster with IGF-Ⅱ firstly and then cluster with IGF-Ⅰcomposing IGF family, and finally IGF family clustered in the same branch with Insulin. Tissue distribution shows that IGF-Ⅲ was widely expressed in heart, gonad and brain of P.diacanthus, especially highest in gonad. Quantitative real-time PCR analysis reveals that the expression of IGF-Ⅲ in testis was significantly higher in developing phase than that in the others, showing a significant decline. The expression of IGF-Ⅲ in ovary of maturation phase was also remarkably higher than that in other periods. Besides, the expression of IGF-Ⅲ in forebrain, midbrain and hindbrain in female fish in mature phase was higher than that in other periods. However, IGF-Ⅲ expression pattern was different in male fish. It is suggested that IGF-Ⅲ might be important for the development of testis and maturation of ovary, and forebrain might involve in the feedback regulation of IGF-Ⅲ expression in gonad.

     

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