Abstract：
FMRFamide, a member of the neuropeptide family, is involved in numerous physiological processes. FMRFamide-activated sodium channels (FaNaCs) are a family of non-voltage-gated, amiloride-sensitive, Na+-selective channels triggered by the neuropeptide FMRFamide. In the present study, the full-length cDNA of the FaNaC receptor of Sepiella japonica (SjFaNaC) was cloned. The cDNA of SjFaNaC was 3004 bp long with an open reading frame (ORF) of 1812 bp, encoding 603 amino acid residues with no signal peptide at the N-terminus. Sequence analysis indicated that SjFaNaC shared a high identity with other cephalopods FaNaCs and formed a sister clade with bivalves. The protein structure was predicted using SWISS-MODEL with AcFaNaC as the template. Quantitative real-time PCR (qRT-PCR) revealed that SjFaNaC transcripts were highly expressed in both female and male reproductive organs, as well as in the optic lobe and brain of the central nervous system (CNS). Results of in situ hybridisation (ISH) showed that SjFaNaC mRNA was mainly distributed in the medulla and deep retina of the optic lobe and in both the supraesophageal and subesophageal masses of the brain. Subcellular localisation indicated that the SjFaNaC protein was localised intracellularly and on the cell surface of HEK293T cells. In summary, these findings may lay the foundation for future exploration of the functions of SjFaNaC in cephalopods.
摘要：
FMRFamide,神经肽家族的一员,参与许多生理过程。FMRFamide激活的钠通道（FaNaC）是一个非电压门控的家族，阿米洛利敏感，由神经肽FMRFamide触发的Na+选择性通道。在本研究中,克隆了日本SepiellaFaNaC受体（SjFaNaC）的全长cDNA。SjFaNaC的cDNA长3004bp,开放阅读框(ORF)为1812bp,编码603个氨基酸残基，N端无信号肽。序列分析表明，SjFaNaC与其他头足类动物FaNaC具有很高的同一性，并与双壳类动物形成了姐妹进化枝。使用以AcFaNaC为模板的SWISS-MODEL预测蛋白质结构。实时定量PCR（qRT-PCR）显示，SjFaNaC转录本在女性和男性生殖器官中均高表达，以及中枢神经系统（CNS）的视叶和大脑。原位杂交（ISH）结果表明，SjFaNaCmRNA主要分布在视叶的延髓和深视网膜以及脑的食道上和食道下肿块中。亚细胞定位表明SjFaNaC蛋白位于HEK293T细胞的细胞内和细胞表面。总之,这些发现可能为将来探索SjFaNaC在头足类动物中的功能奠定基础。
