Page 37 - 《渔业研究》2026年第2期

180 渔 业 研 究 第 48 卷

Characteristics of the HSPA8 gene in Amphiprion ephippium and its response
to transportation stress

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YU Dapeng ，LIANG Renhao ，CAI Jia ，HUANG Xiaolin ，YANG Yukai ，HUANG Zhong ，LI Tao 3,4*
(1. Shenzhen Institute of Guangdong Ocean University, Shenzhen 518121, China;
2. Guangzhou University, Guangzhou 510006, China;
3. Shenzhen Base of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shenzhen 518121, China;
4. Key Laboratory of South China Sea Fishery Research Exploitation & Utilization, Ministry of Agriculture and Rural
Affairs/Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, South China Sea Fisheries Research Institute,
Chinese Academy of Fishery Sciences, Guangzhou 510300, China)
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.
Key words: Amphiprion ephippium; HSPA8 gene; sequence analysis; transport stress; expression analysis