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长期低氧胁迫下红鳍东方鲀鳔的转录组分析
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Abstract:
水中的溶解氧(Dissolved Oxygen, DO)水平影响着鱼类的生存、繁殖、生长和发育。缺氧会对水产养殖业的发展，生物多样性以及生态系统的功能产生影响。因此，本研究以红鳍东方鲀(Takifugu ru-bripes)为研究对象，对常氧(7.5 ± 0.5 mg/L)与低氧(2.5 ± 0.5 mg/L，10天)环境下的红鳍东方鲀的鳔组织进行了转录组测序分析，以探究其应对低氧胁迫时的分子调控机制。随之，筛选出差异表达基因并对其进行生物信息学分析。同时，进一步采用qRT-PCR进行了验证。结果显示，在红鳍东方鲀的鳔组织中共鉴定到了536个差异基因，包括493个上调基因和43个下调基因。KEGG富集结果表明，钙信号传导途径，环磷酸腺苷(cAMP)通路，胰岛素分泌三个信号通路通过调整机体内离子转运，在机体适应低氧环境、维持细胞存活中发挥了重要作用。此外，GSEA富集分析表明，红鳍东方鲀还通过下调DNA复制，细胞周期等耗能的生物过程来抑制细胞正常分裂和生长，从而减少低氧环境中的能量消耗。最后，我们随机挑选了转录组数据中的6个基因进行qRT-PCR验证，结果与转录组数据分析相一致，证实了转录组测序结果的可靠性。本研究结果不仅丰富了红鳍东方鲀响应低氧胁迫的分子调控机制，同时为探究鱼类的低氧适应机制提供了理论依据。
Dissolved oxygen (DO) in water affects the fish’s survival, reproduction, growth, and development. Hypoxia can significantly impact the development of aquaculture, biodiversity, and ecosystem function. Therefore, in this study, the transcriptome of Takifugu rubripes was sequenced to investigate the molecular regulatory mecha-nisms involved in the response to chronic hypoxic stress in the swimbladder under normoxic (7.5 ± 0.5 mg/L) and hypoxic (2.5 ± 0.5 mg/L, 10 days) conditions. Concomitantly, differentially expressed genes (DEGs) were identified and bioinformatics analysis of the DEGs was performed. At the same time, qRT-PCR validation was also carried out. The results showed that a total of 536 DEGs were identified in the T. rubripes swimbladder, including 493 up-regulated genes and 43 down-regulated genes. According to the KEGG enrichment analysis, the calcium signaling pathway, cAMP signaling pathway, and insulin secretion played an important role in adapting to the hypoxic environment and maintaining cell survival by regulating ion transport in the organism. In addition, GSEA en-richment analysis showed that T. rubripes could inhibit cell division and growth via down-regulating energy-consuming biological processes to reduce energy consumption in hypoxic environments, including DNA replication and cell cycle. Finally, 6 genes were randomly selected for qRT-PCR validation and the results proved to be consistent with the transcriptome data analysis, which confirmed the reliability of the transcriptome data. The results of this study not only en-riched the molecular regulatory mechanism of T. rubripes in response to hypoxic stress, but also provided a theoretical basis for exploring the hypoxia adaptation mechanism of fish.
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