Muncaster Simon, Kraakman Kirsty, Gibbons Olivia, Mensink Koen, Forlenza Maria, Jacobson Gregory, Bird Steve
Marine and Environmental Group, School of Applied Science, Bay of Plenty Polytechnic, Tauranga, New Zealand.
Molecular Genetics, School of Science, Faculty of Science and Engineering, University of Waikato, Private Bag 3105, Hamilton 3240, New Zealand.
Dev Comp Immunol. 2018 Mar;80:67-80. doi: 10.1016/j.dci.2017.04.014. Epub 2017 Apr 19.
A number of Seriola species are currently farmed or being investigated as future aquaculture species in countries around the world. However they face a number of issues and limitations which will need to be overcome to ensure future stability and growth, one of which are disease outbreaks. Despite this, very little has been done to understand the immune system of Seriola species and very few immune genes have been characterised. Antimicrobial peptides (AMP) are naturally occurring low molecular weight polypeptides that play a major role in an organism's immune system and act effectively as a first line of defence. This investigation isolates the full length cDNA sequences of two AMP's, piscidin and hepcidin from the yellowtail kingfish (Seriola lalandi). The full-length cDNA of the piscidin gene encodes a 65 amino acid prepropeptide, containing a 25-residue peptide, predicted to form an amphipathic helix-loop-helix structure. Phylogenetic analysis using fish piscidin sequences, showed that this AMP is only found in bony fish within the Acanthomorpha clade and that a possible three groups within the piscidin family exists, with S. lalandi belonging to a particular group. The full-length cDNA of the hepcidin gene encodes a 90 amino acid preprohepcidin, which contains a typical RX(R/K)R motif for cleavage of the mature peptide which comprises of eight conserved cysteine residues. Phylogenetic analysis of known vertebrate hepcidin antimicrobial peptide (HAMP) sequences, shows sequences from the Neoteleostei clade of bony fish form two very separate groups, HAMP1 and HAMP2, with the S. lalandi hepcidin gene grouped with the HAMP1 sequences. HAMP2 sequences are found to have multiple copies within fish and genome analysis showed very clearly that these two groups of genes are located on separate regions on the genome, with the multiple HAMP2 copies formed from tandem gene duplications. Lastly, using qPCR the expression of the S. lalandi piscidin gene within healthy fish was highest within, spleen and gills and lowest in liver, whereas hepcidin was highest in the liver with little or no expression in the spleen and gills.
目前,世界上许多国家都在养殖或研究一些黄尾鰤属鱼类,将其作为未来的水产养殖品种。然而,它们面临着一些问题和限制,为确保未来的稳定和发展,这些问题都需要克服,其中之一就是疾病爆发。尽管如此,人们对黄尾鰤属鱼类免疫系统的了解却非常少,仅有极少数免疫基因得到了鉴定。抗菌肽(AMP)是天然存在的低分子量多肽,在生物体免疫系统中发挥着重要作用,是有效的第一道防线。本研究从黄尾鰤(Seriola lalandi)中分离出两种抗菌肽——杀鱼菌素和铁调素的全长cDNA序列。杀鱼菌素基因的全长cDNA编码一个65个氨基酸的前原肽,包含一个25个残基的肽段,预计形成两亲性螺旋-环-螺旋结构。利用鱼类杀鱼菌素序列进行的系统发育分析表明,这种抗菌肽仅在棘鳍总目分支内的硬骨鱼中发现,并且杀鱼菌素家族内可能存在三个组,黄尾鰤属于特定的一组。铁调素基因的全长cDNA编码一个90个氨基酸的前原铁调素肽,其中包含一个典型的RX(R/K)R基序,用于切割由八个保守半胱氨酸残基组成的成熟肽。对已知脊椎动物铁调素抗菌肽(HAMP)序列的系统发育分析表明,硬骨鱼新真骨鱼总目分支的序列形成两个非常不同的组,即HAMP1和HAMP2,黄尾鰤铁调素基因与HAMP1序列归为一组。研究发现,HAMP2序列在鱼类中有多个拷贝,基因组分析非常清楚地表明,这两组基因位于基因组的不同区域,多个HAMP2拷贝是由串联基因重复形成的。最后,使用定量PCR技术检测发现,健康黄尾鰤体内杀鱼菌素基因在脾脏和鳃中的表达最高,在肝脏中的表达最低,而铁调素在肝脏中的表达最高,在脾脏和鳃中几乎不表达或不表达。