合浦珠母贝章鱼胺受体(OAR)基因的克隆与表达分析

Molecular cloning and expression analysis of octopamine receptor gene in pearl oyster (Pinctada fucata)

  • 摘要: 章鱼胺受体(octopamine receptor,OAR)广泛存在于无脊椎动物中,具有调节神经肌源性节奏收缩、调节cAMP水平、参与磷酸化途径等多种重要功能,但其在贝类中的研究却很少。该研究克隆了合浦珠母贝(Pinctada fucata)OAR基因(pfOAR)的cDNA全长序列,比较分析了氨基酸序列的同源性,并采用荧光定量PCR技术检测了该基因在不同组织和幼体不同发育阶段的表达差异以及贝壳损伤后该基因在外套膜中表达水平的变化。结果显示:pfOAR的cDNA全长1 890 bp,开放阅读框1 659 bp,编码552个氨基酸,有7个跨膜结构域,具有G蛋白偶联受体的共性。pfOAR在6个被检测的组织中均有表达,且在外套膜中的表达水平显著高于其他组织;幼体发育过程中pfOAR表达水平逐渐升高,变态期的表达水平显著高于其他时期;人为损伤贝壳36 h后,pfOAR的表达水平显著升高。这些结果表明pfOAR在合浦珠母贝中可能具有多样化的功能,参与了贝壳的形成和修复,为进一步开展pfOAR的功能研究奠定了重要基础。

     

    Abstract: Octopamine receptor (OAR) is common in invertebrates and plays an important role in invertebrate life, including adjusting nerve muscle rhythm contraction and the cAMP level, participating in phosphorylation pathways, etc.. However, there is little research on the OAR gene in mollusks. In this study, we cloned an OAR gene from pearl oyster (Pinctada fucata) to investigate the characteristics of the cDNA and deduced amino acid sequences, relative expression levels in different tissues and development stages using qPCR, as well as the relative expression levels after shell damage. The results show that the full length cDNA of pfOAR was 1 890 bp, with 1 659 open reading frame (ORF), encoding 552 amino acids. The pfOAR had common features of the vast family of G protein-coupled receptors including seven hydrophobic transmembrane domains. The qPCR analysis reveals ubiquitous expression of pfOAR mRNA in all six tissues, with significantly high expression level in mantle tissue and at metamorphosis stage during larval development. Furthermore, the relative expression level reached the maximum in 36 h after shell notching. It is conluded that pfOAR might play various roles in P.fucata, including possible involvement in the formation and repairment of the shell, which provides references for further functional study of pfOAR.

     

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