渔业研究2026，Vol.48Issue(2)：169-180,12.DOI:10.14012/j.jfr.2025087
大眼双锯鱼HSPA8基因特征及其对运输胁迫的响应
Characteristics of the HSPA8 gene in Amphiprion ephippium and its response to transportation stress
摘要
Abstract
[Background]Amphiprion ephippium is a significant ornamental marine species.Transportation stress,a common environmental stress during fish transport,may adversely affect its immune systems.[Object-ive]This study aims to investigate the biological characteristics of heat shock protein 8(HSPA8)in A.ephippi-um and its expression patterns under transportation conditions.[Methods]Using the liver tissue of A.ephippi-um,polymerase chain reaction(PCR)was employed to amplify and clone the coding sequences(CDS)of the HSPA8 gene.Bioinformatics analysis was conducted to evaluate its biological properties and structural features,along with detection of relative expression levels across different tissues.A transportation-induced stress model was constructed,and quantitative real-time PCR(qPCR)was used to monitor HSPA8 gene expression changes in various tissues at different transportation time points.[Results]The CDS region of the HSPA8 gene in A.ephippium contained 1 953 bp,encoding 650 amino acid residues with a molecular weight of approximately 71.26 kDa and a theoretical isoelectric point of 5.22.The open reading frame(ORF)and protein sequence length of the HSPA8 gene in A.ephippium were identical to those of A.ocellaris.The HSPA8 gene ORF of A.ephippium differed by 16 base pairs,with a sequence similarity of 99.18%,but the protein sequences were completely identical.Phylogenetic analysis showed that A.ephippium was closely related to A.ocellaris.Sub-sequent analyses revealed the absence of signal peptide sequences and transmembrane domains,but the protein contained three typical structural domains of HSP70s proteins.qPCR analysis of HSPA8 gene tissue-specific ex-pression demonstrated its presence in all 10 tested tissues(skin,muscle,spleen,gill,liver,heart,intestine,brain,kidney,and eye).The gene showed highest relative expression in the eye,followed by kidney and brain tissues,with lower levels in heart and gill tissues,and the lowest level in skin.Under transport stress conditions,HSPA8 gene expression in the spleen was significantly upregulated at 6 and 12 hours,while intestine upregulation was observed at 6 hours,but not at other time points showing significance.Liver HSPA8 gene expression remained significantly lower than the control groups at 6,12,24,and 48 hours.Skin HSPA8 gene expression showed highly significant elevation at 24 hours compared to the control group,with no significant changes observed at other time points.[Conclusion]The HSPA8 gene participates in stress responses in immune tissues in A.ephip-pium,making it a reliable indicator of whether immunosuppression occurs during stress.The above results provide a reference for further study of the immune regulatory function of the HSPA8 gene under transport stress.关键词
大眼双锯鱼/HSPA8 基因/序列分析/运输胁迫/表达分析Key words
Amphiprion ephippium/HSPA8 gene/sequence analysis/transport stress/expression analysis分类
农业科技引用本文复制引用
喻大鹏,梁仁豪,蔡佳,黄小林,杨育凯,黄忠,李涛..大眼双锯鱼HSPA8基因特征及其对运输胁迫的响应[J].渔业研究,2026,48(2):169-180,12.基金项目
海南省科技专项(ZDYF2022XDNY349) （ZDYF2022XDNY349）
深圳市规划与自然资源局农业发展专项资金(渔业类)农业高新技术项目(2023-222) （渔业类）
