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利用转录组分析揭示 4-壬基酚暴露下凡纳滨对虾肝胰腺调控机制的新见解。

New insights into the regulation mechanism of Pacific white shrimp (Litopenaeus vannamei) hepatopancreas under 4-nonylphenol exposure using transcriptome analysis.

机构信息

Hainan Provincial Key Laboratory for Functional Components Research and Utilization of Marine Bio-resources, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences & Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province, Hainan Institute of Tropical Agricultural Resources, Haikou, 571101, PR China.

Hainan Provincial Key Laboratory for Functional Components Research and Utilization of Marine Bio-resources, Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences & Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province, Hainan Institute of Tropical Agricultural Resources, Haikou, 571101, PR China.

出版信息

Fish Shellfish Immunol. 2023 Oct;141:109050. doi: 10.1016/j.fsi.2023.109050. Epub 2023 Sep 2.

Abstract

4-Nonylphenol (4-NP) is one of the common endocrine-disrupting chemicals (EDCs) in estuaries and coastal zones, which can exert detrimental effects on the physiological function of aquatic organisms. However, the molecular response triggered by 4-NP remains largely unknown in Pacific white shrimp (Litopenaeus vannamei). In this study, transcriptomic analysis was performed to investigate the underlying mechanisms of 4-NP toxicity in the hepatopancreas of L. vannamei. Nine RNA-Seq libraries were generated from L. vannamei at 0 h, 24 h, and 48 h following exposure to 4-NP. Compared with 0 h vs 24 h, 962 up- and 463 down-regulated differentially expressed genes (DEGs) were identified, indicating that many genes in L. vannamei were induced to resist adverse circumstances by 4-NP exposure. In contrast, 902 up- and 1027 down-regulated DEGs were revealed in the comparison of 0 h vs 48 h, demonstrating that prolonged exposure to the stress from 4-NP resulted in more inhibited genes. To validate the accuracy of the transcriptome data, eight DEGs were selected for quantitative real-time polymerase chain reaction (qRT-PCR), which were consistent with the RNA-Seq results. Through KEGG pathway enrichment analysis, three specific pathways related to hormonal effects and endocrine function of L. vannamei were enriched significantly, including tyrosine metabolism, insect hormone biosynthesis, and melanogenesis. After 4-NP stress, genes involved in tyrosine metabolism (Tyr) and melanogenesis pathway (AC, CBP, Wnt, Frizzled, Tcf, and Ras) were induced to promote melanin pigment to help shrimp resist adverse environments. In the insect hormone biosynthesis, ALDH, CYP15A1, CYP15A1/C1, and JHE genes were activated to synthesize juvenile hormone (JH), while Spook, Phm, Sad, and CYP18A1 were induced to generate molting hormone. There is an enhanced interaction between the molting hormone and JH, with JH playing a dominant role and maintaining its "classic status quo action". Our study demonstrated that 4-NP exposure led to impairments of biological functions in L. vannamei hepatopancreas. The genes and pathways identified provide novel insights into the molecular mechanisms underlying 4-NP toxicity effects in prawns and enrich the information on the toxicity mechanism of crustaceans in response to EDCs exposure.

摘要

壬基酚(4-NP)是港湾和沿海地区常见的内分泌干扰化学物质(EDCs)之一,它会对水生生物的生理功能产生有害影响。然而,4-NP 引发的分子反应在凡纳滨对虾(Litopenaeus vannamei)中仍知之甚少。在这项研究中,进行了转录组分析,以研究 4-NP 对凡纳滨对虾肝胰腺毒性的潜在机制。从暴露于 4-NP 的凡纳滨对虾中,在 0 h、24 h 和 48 h 时生成了 9 个 RNA-Seq 文库。与 0 h 与 24 h 相比,鉴定出 962 个上调和 463 个下调的差异表达基因(DEGs),表明 4-NP 暴露诱导了凡纳滨对虾中的许多基因抵抗不利环境。相比之下,在 0 h 与 48 h 的比较中,揭示了 902 个上调和 1027 个下调的 DEGs,表明长期暴露于 4-NP 应激导致更多受抑制的基因。为了验证转录组数据的准确性,选择了 8 个 DEGs 进行定量实时聚合酶链反应(qRT-PCR),结果与 RNA-Seq 结果一致。通过 KEGG 途径富集分析,发现三个与凡纳滨对虾激素作用和内分泌功能相关的特定途径显著富集,包括酪氨酸代谢、昆虫激素生物合成和黑色素生成。在 4-NP 应激后,参与酪氨酸代谢(Tyr)和黑色素生成途径(AC、CBP、Wnt、Frizzled、Tcf 和 Ras)的基因被诱导以促进黑色素色素帮助虾类抵抗不利环境。在昆虫激素生物合成中,ALDH、CYP15A1、CYP15A1/C1 和 JHE 基因被激活以合成保幼激素(JH),而 Spook、Phm、Sad 和 CYP18A1 被诱导生成蜕皮激素。蜕皮激素和 JH 之间的相互作用增强,JH 起主导作用并保持其“经典现状作用”。我们的研究表明,4-NP 暴露导致凡纳滨对虾肝胰腺生物功能受损。鉴定出的基因和途径为虾类中 4-NP 毒性作用的分子机制提供了新的见解,并丰富了甲壳类动物对 EDCs 暴露的毒性机制信息。

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