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比较转录组揭示了分子水平下日本沼虾(Macrobrachium nipponense)对硫化物毒性的反应。

Comparative transcriptome reveals the response of oriental river prawn (Macrobrachium nipponense) to sulfide toxicity at molecular level.

机构信息

Department of Aquaculture Research Lab, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, China.

Ministry of Education Key Laboratory of Marine Genetics and Breeding, College of Marine Science, Ocean University of China, Qingdao, 266003, China.

出版信息

Aquat Toxicol. 2021 Jan;230:105700. doi: 10.1016/j.aquatox.2020.105700. Epub 2020 Nov 28.

Abstract

Aquatic environmental pollutants have various impacts on aquaculture. Specifically, sulfide has been established as being toxic to aquatic animals including the oriental river prawn Macrobrachium nipponense. In response, the hepatopancreas has been broadly studied, as it plays a pivotal role in arthropod nutrient digestion and absorption, energy supply, and organ development as well as in crustacean immunity. However, the underlying molecular mechanisms of hepatopancreas's response to sulfide toxicity are still poorly understand. Herein, we used Nova-seq 6000 platform to conduct a comparative transcriptome analysis of gene expression profiles in the hepatopancreas of M. nipponense, while it was under the influence of a semi-lethal sulfide concentration (3.20 mg/L at 48 h). A total of 139 million raw reads were obtained, in which 67,602 transcripts were clustered into 37,041 unigenes for further analysis. After constant sulfide exposure for 48 h, 235 differentially expressed genes, i.e., DEGs (151 up-regulated and 84 down-regulated) were identified in the sulfide treatment group (TGHP) compared with the control group (CGHP). We used GO and KEGG databases to annotate all the DEGs into 1180 functions and 123 pathways, respectively. The metabolic pathways included proximal tubule bicarbonate reclamation, sulfur metabolism, glycolysis and gluconeogenesis, and the TCA cycle; while immune-related pathways contained Ras, Rap1, focal adhesion and platelet activation. Additionally, apoptosis-involved pathways e.g., lysosome, also exhibited remarkable alteration in the presence of sulfide stress. Notably, responses to external stimuli and detoxification genes- such as GSKIP, CRT2, APOD, TRET1, CYP4C3 and HR39- were significantly altered by the sulfide stress, indicating that significant toxicity was transferred through energy metabolism, growth, osmoregulatory processes and immunity. Finally, we demonstrated that in the present of sulfide stress, M. nipponense altered the expression of detoxification- and extracellular stimulation-related genes, and displayed positive resistance via tight junction activation and lysosome pathways. The results of these novel experiments shed light on the hepatopancreas's molecular response to sulfide stress resistance and the corresponding adaptation mechanism; and enable us to identify several potential biomarkers for further studies.

摘要

水生环境污染物对水产养殖有多种影响。具体来说,已经确定硫化物对包括日本沼虾(Macrobrachium nipponense)在内的水生动物具有毒性。因此,人们广泛研究了肝胰腺,因为它在节肢动物的营养消化和吸收、能量供应以及器官发育和甲壳类动物的免疫中起着关键作用。然而,肝胰腺对硫化物毒性反应的潜在分子机制仍知之甚少。在此,我们使用 Nova-seq 6000 平台对日本沼虾肝胰腺在受到半致死硫化物浓度(48 小时时为 3.20mg/L)影响时的基因表达谱进行了比较转录组分析。共获得 1.39 亿条原始读数,其中 67602 条转录物聚类为 37041 条非冗余基因用于进一步分析。经过 48 小时持续硫化物暴露后,与对照组(CGHP)相比,硫化物处理组(TGHP)中鉴定出 235 个差异表达基因(151 个上调和 84 个下调)。我们使用 GO 和 KEGG 数据库分别将所有 DEGs 注释为 1180 种功能和 123 种途径。代谢途径包括近端肾小管碳酸氢盐回收、硫代谢、糖酵解和糖异生以及三羧酸循环;而免疫相关途径包括 Ras、Rap1、粘着斑和血小板激活。此外,溶酶体等涉及细胞凋亡的途径也在存在硫化物胁迫时发生明显改变。值得注意的是,对外界刺激和解毒基因的反应,如 GSKIP、CRT2、APOD、TRET1、CYP4C3 和 HR39,在受到硫化物胁迫时明显改变,表明通过能量代谢、生长、渗透压调节过程和免疫传递了显著的毒性。最后,我们证明在硫化物胁迫下,日本沼虾改变了解毒和细胞外刺激相关基因的表达,并通过紧密连接激活和溶酶体途径显示出积极的抗性。这些新实验的结果揭示了肝胰腺对硫化物胁迫抗性的分子反应及其相应的适应机制;并使我们能够确定几个潜在的生物标志物进行进一步研究。

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