金鱼(Carassius auratus L.)体内 Caspase-8 的分子克隆和功能鉴定。

IF 4.1 2区 农林科学 Q1 FISHERIES
Ziqi Ban, Xinyuan Yue, Kejing Huang, Lu Yuan, Suming Zhou, Jianhu Jiang, Chenjie Fei, Jiasong Xie
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引用次数: 0

摘要

细胞凋亡是一种程序性细胞死亡,对于维持体内环境的平衡至关重要。Caspase-8是一种启动caspase,在凋亡过程中激活caspase级联过程中起着关键作用。本研究克隆并表达了金鱼的 Caspase-8,旨在研究 Caspase-8 在鱼类对细菌感染(特别是由嗜水气单胞菌引起的感染)的免疫反应中的作用。Gf-Casp8 基因由 1,425 个碱基对组成,编码 474 个氨基酸的蛋白质,分子量为 54.55 kDa。实时定量 PCR 分析显示,Gf-Casp8 在脾脏和肾脏中的表达量较高,而在肌肉和心脏中的表达量较低。此外,肾脏白细胞在受到蚜蝇和 LPS 刺激后,Gf-Casp8 的表达明显上调。免疫后,肾脏中 Gf-Casp8 的 mRNA 和蛋白质水平都明显升高。亚细胞定位分析表明,Gf-Casp8 定位于细胞质和细胞核中。此外,流式细胞术分析表明,过量表达 Gf-Casp8 能明显增强 HEK293T 细胞的凋亡。这些结果突显了 Gf-Casp8 在调节金鱼细胞凋亡和抗菌免疫反应中的关键功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Molecular cloning and functional characterization of Caspase-8 in goldfish (Carassius auratus L.).

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.

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来源期刊
Fish & shellfish immunology
Fish & shellfish immunology 农林科学-海洋与淡水生物学
CiteScore
7.50
自引率
19.10%
发文量
750
审稿时长
68 days
期刊介绍: Fish and Shellfish Immunology rapidly publishes high-quality, peer-refereed contributions in the expanding fields of fish and shellfish immunology. It presents studies on the basic mechanisms of both the specific and non-specific defense systems, the cells, tissues, and humoral factors involved, their dependence on environmental and intrinsic factors, response to pathogens, response to vaccination, and applied studies on the development of specific vaccines for use in the aquaculture industry.
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