厚壳贻贝5-羟色胺2A受体(5-HT2AR)基因克隆和时空表达

5-HT2AR gene clone and temporal-spatial expression in the mussel Mytilus coruscus

  • 摘要: 为探究5-羟色胺2A受体(5-HT 2A receptor, 5-HT2AR)基因在海水贝类生长发育中的作用,本研究通过RACE技术和实时荧光定量PCR(qRT-PCR)克隆了厚壳贻贝5-HT2AR 基因的cDNA全长,并分析该基因的时空表达。结果显示,5-HT2AR基因全长2 636 bp,开放阅读框(ORF)2 124 bp,共编码707个氨基酸。序列分析结果显示该序列与人、小鼠、斑马鱼、长牡蛎和虾夷扇贝等物种分别具有45%、45%、48%、49%和67%的同源性。厚壳贻贝雌雄成体各组织和器官中均有5-HT2AR基因表达,雄性中的鳃表达量最高,而在雌性鳃、外套膜和性腺中的表达稍高于其他组织和器官;推测该基因可能与厚壳贻贝的摄食、对外界环境的感知及促进卵母细胞成熟有关。5-HT2AR基因在厚壳贻贝各发育阶段均有表达,且稚贝的表达量为眼点幼虫的1.4倍,推测5-HT2AR基因可能参与了调控厚壳贻贝幼虫的生长发育过程。本研究为进一步了解5-HT基因家族在双壳贝类中的功能奠定了基础。

     

    Abstract: To understand the role of serotonin 5-HT2A receptor (5-HT2AR) gene in the growth and development of the mussel Mytilus coruscus, we cloned the 5-HT2AR gene cDNA full length of M. coruscus through RACE technology and analyzed the temporal and spatial expression of this gene via the real-time fluorescent quantitative PCR (qRT-PCR). The full length of 5-HT2AR gene was 2 636 bp and 2 124 bp of open reading frame (ORF) encoding 707 amino acids. Homologous analysis of the amino acids of 5-HT2AR gene in M. coruscus shared high similarity with Homo sapiens (45%), Mus musculus (45%), Danio rerio (48%), Crassostrea gigas (49%) and Mizuhopecten yessoensis (67%), respectively. This study showed that 5-HT2AR mRNA expression was detected in all tissues and the highest expression was found in gill and female gonad, indicating that 5-HT2AR gene may play an important role in perception of the external environment and promotion of mature oocytes. The 5-HT2AR mRNA was expressed at all developmental stages, suggesting that 5-HT2AR gene may be involved in the regulation of the growth and development. Taken together, our findings may provide new insights into clarifying the function of 5-HT gene family in molluscs.

     

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