Molecular cloning and expression analysis of rainbow trout (Oncorhynchus mykiss) interleukin-10 cDNAs
Molecular cloning and expression analysis of rainbow trout (Oncorhynchus mykiss) interleukin-10 cDNAs
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DOI：
10.1016/j.fsi.2004.08.004
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发表时间：
2005-04-01
影响因子：
4.7
通讯作者：
Nakanishi, T
中科院分区：
文献类型：
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作者：
Inoue, Y;Kamota, S;Nakanishi, T
本研究利用河豚鱼和鲤鱼IL-10基因序列，从虹鳟鱼中分离IL-10，并对其结构进行分析。同时检测了虹鳟鱼不同组织中IL-10 mRNA的表达。为了减少混淆的可能性，目前的结果与个体之间的差异，纯合同基因虹鳟鱼，以尽量减少个体之间的遗传变异。结果表明，虽然虹鳟鱼IL-10基因与人IL-10基因同源性不高，但其生物活性所必需的位点和基序以及三级结构都很保守。虹鳟鱼IL-10成熟肽中含有4个半胱氨酸残基（Cys-32、82、132和138），这4个半胱氨酸残基在所有脊椎动物IL-10肽中都是保守的，但在所有鱼类IL-10中都存在2个额外的半胱氨酸残基（Cys-28和33）。虹鳟鱼IL-10也具有全部6个α-螺旋，脊椎动物中最保守的区域是螺旋C、DE环和螺旋F。虹鳟鱼IL-10有一个潜在的糖基化位点。该基因在鳃中强表达，在头肾、脾和肠中弱表达。IL-10在正常组织中的表达模式与河豚鱼（在肾脏和肝脏中弱表达）和鲤鱼（在头肾和肠中强表达）的表达模式有很大不同。虹鳟鱼IL-10在脾和肾中的高表达与LPS刺激可以归因于这些组织中的吞噬细胞的存在。在虹鳟鱼与LPS刺激的表达模式似乎是一致的，它是一种炎症细胞因子。
In the present study, the sequences of the fugu and carp IL-10 genes were used to isolate IL-10 from rainbow trout and analysed its structure. IL-10 mRNA expression in different tissues of rainbow trout was also examined. To reduce the possibility of confounding the present results with differences among individuals, homozygous isogenetic rainbow trout were used to minimize the genetic variation among individuals. Results showed that although the IL-10 gene of the rainbow trout was not highly homologous with human IL-10, the sites and motifs essential for the bioactivity and tertiary structure were well conserved. The predicted mature peptide of rainbow trout IL-10 contained four cysteine residues (Cys-32, 82, 132 and 138) that were conserved in all vertebrate IL-10 peptides, although two extra cysteine residues (Cys-28 and 33) were present in all fish IL-10s. Rainbow trout IL-10 also possessed all six α-helices and the most conserved region were helix C, the DE loop and helix F in vertebrates. Rainbow trout IL-10 had one potential glycosylation site. The gene was strongly expressed in the gill and weakly expressed in the head kidney, spleen and intestine. In normal tissues, the IL-10 expression pattern was quite different from the expression patterns of fugu (weakly expressed in the kidney and liver) and carp (strongly expressed in the head kidney and intestine). The high expression of rainbow trout IL-10 in spleen and kidney following stimulation with LPS can be attributed to the presence of phagocytic cells in these tissues. The expression pattern in rainbow trout stimulated with LPS seemed consistent with its being an inflammatory cytokine.