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整合转录组和 miRNA 测序分析表明,低氧应激会导致遗传改良养殖罗非鱼(GIFT,Oreochromis niloticus)鳃中的免疫和代谢紊乱。

Integrated transcriptome and miRNA sequencing analyses reveal that hypoxia stress induces immune and metabolic disorders in gill of genetically improved farmed tilapia (GIFT, Oreochromis niloticus).

机构信息

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, China.

Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, China.

出版信息

Fish Shellfish Immunol. 2023 Aug;139:108909. doi: 10.1016/j.fsi.2023.108909. Epub 2023 Jun 21.

Abstract

The survival and growth of fish are significantly impacted by a hypoxic environment (low dissolved oxygen). In this study, we compared tissue structure, physiological changes, and mRNA/miRNA transcriptome, in gills of genetically improved farmed tilapia (GIFT, Oreochromis niloticus) between the hypoxic group (DO: 0.55 mg/L, HG) and the control group (DO: 5 mg/L, CG). The results showed that the gill filaments in the hypoxic group showed curling, engorgement, and apoptotic cells increased, and that exposure for 96 h resulted in a reduction in the antioxidant capacity. We constructed and sequenced miRNA and mRNA libraries from gill tissues of GIFT at 96 h of hypoxia stress. Between the HG and CG, a total of 14 differentially expressed (DE) miRNAs and 1557 DE genes were obtained. GO and KEGG enrichment showed that DE genes were mainly enriched in immune and metabolic pathways such as natural killer cell mediated cytotoxicity, steroid biosynthesis, primary immunodeficiency, and synthesis and degradation of ketone bodies. Based on the results of mRNA sequencing and screening for miRNA-mRNA pairs, we selected and verified six DE miRNAs and their probable target genes. The sequencing results were consistent with the qRT-PCR validation results. The result showed that under hypoxia stress, the innate immune response was up-regulated, and the adaptive immune response was down-regulated in the gill of GIFT. The synthesis of cholesterol in gill cells is reduced, which is conducive to the absorption of solvent oxygen. These findings offer fresh information about the processes of fish adaptation to hypoxic stress.

摘要

鱼类的生存和生长受到低氧环境(溶解氧低)的显著影响。在这项研究中,我们比较了遗传改良的罗非鱼(奥利亚罗非鱼)在缺氧组(DO:0.55mg/L,HG)和对照组(DO:5mg/L,CG)的组织结构、生理变化和 mRNA/miRNA 转录组。结果表明,缺氧组的鳃丝出现卷曲、充血和凋亡细胞增加,暴露 96h 后抗氧化能力降低。我们从缺氧应激 96h 的 GIFT 鳃组织构建并测序了 miRNA 和 mRNA 文库。在 HG 和 CG 之间,共获得了 14 个差异表达(DE)miRNA 和 1557 个 DE 基因。GO 和 KEGG 富集显示,DE 基因主要富集在免疫和代谢途径,如自然杀伤细胞介导的细胞毒性、类固醇生物合成、原发性免疫缺陷和酮体的合成和降解。基于 mRNA 测序和 miRNA-mRNA 对的筛选结果,我们选择并验证了六个 DEmiRNA 及其可能的靶基因。测序结果与 qRT-PCR 验证结果一致。结果表明,在缺氧应激下,GIFT 鳃中的先天免疫反应上调,适应性免疫反应下调。鳃细胞中胆固醇的合成减少,有利于溶剂氧的吸收。这些发现为鱼类适应低氧应激的过程提供了新的信息。

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