Molecular cloning and functional characterization of Caspase-8 in goldfish (Carassius auratus L.)

Ziqi Ban; Xinyuan Yue; Kejing Huang; Lu Yuan; Suming Zhou; Jianhu Jiang; Chenjie Fei; Jiasong Xie

doi:10.1016/j.fsi.2024.110090

Molecular cloning and functional characterization of Caspase-8 in goldfish (Carassius auratus L.)

Fish Shellfish Immunol. 2024 Dec 14:157:110090. doi: 10.1016/j.fsi.2024.110090. Online ahead of print.

Authors

Ziqi Ban¹, Xinyuan Yue¹, Kejing Huang¹, Lu Yuan¹, Suming Zhou², Jianhu Jiang³, Chenjie Fei⁴, Jiasong Xie⁵

Affiliations

¹ School of Marine Sciences, Ningbo University, China, Ningbo, Zhejiang, 315211, China.
² School of Marine Sciences, Ningbo University, China, Ningbo, Zhejiang, 315211, China; Key Laboratory of Aquacultural Biotechnology, Ministry of Education, Ningbo University, Ningbo, Zhejiang, 315211, China; Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo, Zhejiang, 315211, China.
³ Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang, 313001, China.
⁴ School of Marine Sciences, Ningbo University, China, Ningbo, Zhejiang, 315211, China; Key Laboratory of Aquacultural Biotechnology, Ministry of Education, Ningbo University, Ningbo, Zhejiang, 315211, China; Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo, Zhejiang, 315211, China. Electronic address: [email protected].
⁵ School of Marine Sciences, Ningbo University, China, Ningbo, Zhejiang, 315211, China; Key Laboratory of Aquacultural Biotechnology, Ministry of Education, Ningbo University, Ningbo, Zhejiang, 315211, China; Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo, Zhejiang, 315211, China. Electronic address: [email protected].

PMID: 39681222
DOI: 10.1016/j.fsi.2024.110090

Abstract

Apoptosis, a form of programmed cell death, is essential for maintaining homeostasis within the internal environment. Caspase-8, an initiator caspase, plays a pivotal role in activating the caspase cascade during the apoptotic process. This study cloned and expressed Caspase-8 from goldfish, aimed to investigate the role of Caspase-8 in the immune response of fish to bacterial infections, specifically those caused by Aeromonas hydrophila. The Gf-Casp8 gene consists of 1425 base pairs, encoding a protein of 474 amino acids with a molecular weight of 54.55 kDa. Quantitative real-time PCR analysis revealed that Gf-Casp8 is highly expressed in the spleen and kidney, with lower expression levels in the muscle and heart. Moreover, Gf-Casp8 expression was significantly upregulated in kidney leukocytes following stimulation with A. hydrophila and LPS. Post-immunization, both mRNA and protein levels of Gf-Casp8 in the kidney were significantly increased. Subcellular localization analysis indicated that Gf-Casp8 is localized in both the cytoplasm and nucleus. In addition, flow cytometry analysis demonstrated that overexpression of Gf-Casp8 significantly enhances apoptosis in HEK293T cells. These results highlight the critical function of Gf-Casp8 in modulating apoptosis and antibacterial immune responses in goldfish.

Keywords: Aeromonas hydrophila; Apoptosis; Caspase-8; Gene expression; Goldfish.