Lu Xing, Xiang Ying, Yang Guohua, Zhang Lang, Wang Hui, Zhong Shan
Key Laboratory of Freshwater Biodiversity Conservation and Utilization of Ministry of Agriculture, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, Hubei, China.
School of Basic Medical Science, Wuhan University, Wuhan 430071, Hubei, China.
Comp Biochem Physiol C Toxicol Pharmacol. 2017 Feb;192:40-49. doi: 10.1016/j.cbpc.2016.12.006. Epub 2016 Dec 7.
Mercury is a widespread toxicant in aquatic environment that can cause deleterious effects on fish. Although a number of mercury-regulated genes have been investigated in adult fish, the transcriptional responses of fish larvae to acute mercury exposure are not well understood. In this study, RNA sequencing was used to examine the transcriptional changes in developing zebrafish larvae under a low concentration of mercuric chloride exposure from 24 to 120hpf. Our initial results showed that a total of 142.59 million raw reads were obtained from sequencing libraries and about 86% of the processed reads were mapped to the reference genome of zebrafish. Differential expression analysis identified 391 up- and 87 down-regulated genes. Gene ontology enrichment analysis revealed that most of the differential expressed genes are closely related to the regulation of cellular process, metabolic process, multicellular organismal process, biological regulation, pigmentation, and response to stimulus. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis demonstrated that antigen processing and presentation was the most significantly enriched pathway. Moreover, we characterized a novel and sensitive mercury-induced ABCB (ATP- binding cassette B subfamily) transporter gene - abcb5. This gene is localized on zebrafish chromosome 16 and contains a 4014bp open-reading frame. The deduced polypeptide is composed of 1337 amino acids and possesses most of functional domains and critical residues defined in human and mouse ABCB5/Abcb5. Functional analysis in vitro demonstrated that overexpression of zebrafish abcb5 gene can significantly decrease the cytotoxicity of mercury in LLC-PK1 cells, implying it is a potential efflux transporter of mercury. Thus, these findings provide useful insights to help further understand the transcriptional response and detoxification ability of zebrafish larvae following acute exposure to mercury.
汞是水生环境中一种广泛存在的有毒物质,会对鱼类造成有害影响。尽管已经在成年鱼类中研究了许多汞调控基因,但鱼类幼体对急性汞暴露的转录反应仍未得到充分了解。在本研究中,利用RNA测序来检测低浓度氯化汞暴露24至120小时后发育中的斑马鱼幼体的转录变化。我们的初步结果显示,从测序文库中总共获得了1.4259亿条原始读数,约86%的处理后读数被映射到斑马鱼的参考基因组。差异表达分析确定了391个上调基因和87个下调基因。基因本体富集分析表明,大多数差异表达基因与细胞过程、代谢过程、多细胞生物过程、生物调控、色素沉着和对刺激的反应的调控密切相关。京都基因与基因组百科全书通路富集分析表明,抗原加工和呈递是最显著富集的通路。此外,我们鉴定了一个新的、对汞敏感的ABCB(ATP结合盒B亚家族)转运蛋白基因——abcb5。该基因位于斑马鱼16号染色体上,包含一个4014bp的开放阅读框。推导的多肽由1337个氨基酸组成,具有人类和小鼠ABCB5/Abcb5中定义的大多数功能域和关键残基。体外功能分析表明,斑马鱼abcb5基因的过表达可显著降低LLC-PK1细胞中汞的细胞毒性,这意味着它是一种潜在的汞外排转运蛋白。因此,这些发现为进一步了解斑马鱼幼体急性暴露于汞后的转录反应和解毒能力提供了有用的见解。