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Spliced leader-based analyses reveal the effects of polycyclic aromatic hydrocarbons on gene expression in the copepod Pseudodiaptomus poplesia
Zhuang, Yunyun1; Yang, Feifei1,2; Xu, Donghui1,4; Chen, Hongju1; Zhang, Huan1,3; Liu, Guangxing1
2017-02-01
Source PublicationAQUATIC TOXICOLOGY
Volume183Pages:114-126
SubtypeArticle
AbstractPolycyclic aromatic hydrocarbons (PAHs) are a group of toxic and carcinogenic pollutants that can adversely affect the development, growth and reproduction of marine organisms including copepods. However, knowledge on the molecular mechanisms regulating the response to PAH exposure in marine planktonic copepods is limited. In this study, we investigated the survival and gene expression of the calanoid copepod Pseudodiaptomus poplesia upon exposure to two PAHs, 1, 2-dimethylnaphthalene (1, 2-NAPH) and pyrene. Acute toxicity responses resulted in 96-h LC50 of 788.98 mu g L-1 and 54.68 mu g L-1 for 1, 2-NAPH and pyrene, respectively. Using the recently discovered copepod spliced leader as a primer, we constructed full-length cDNA libraries from copepods exposed to sublethal concentrations and revealed 289 unique genes of diverse functions, including stress response genes and novel genes previously undocumented for this species. Eighty-three gene families were specifically expressed in PAH exposure libraries. We further analyzed the expression of seven target genes by reverse transcription-quantitative PCR in a time-course test with three sublethal concentrations. These target genes have primary roles in detoxification, oxidative defense, and signal transduction, and include different forms of glutathione S-transferase (GST), glutathione peroxidases (GPX), peroxiredoxin (PRDX), methylmalonate-semialdehyde dehydrogenase (MSDH) and ras-related C3 botulinum toxin substrate (RAC1). Expression stability of seven candidate reference genes were evaluated and the two most stable ones (RPL15 and RPS20 for 1, 2-NAPH exposure, RPL15 and EF1D for pyrene exposure) were used to normalize the expression levels of the target genes. Significant upregulation was detected in GST-T, GST-DE, GPX4, PRDX6 and RAC1 upon 1, 2-NAPH exposure, and GST-DE and MSDH upon pyrene exposure. These results indicated that the oxidative stress was induced and that signal transduction might be affected by PAH exposure in P. poplesia. However, gene upregulation was followed by a reduction in expression level towards 96 h, indicating a threshold value of exposure time that leads to depressed gene expression. Prolonged exposure may cause dysfunction of detoxification and antioxidant machinery in P. poplesia. The transcriptional responses of GST-T, GPX2 and GPX4 upon pyrene exposure were minimal. Our results reveal the different sensitivity of P. poplesia to two PAHs at both the individual and transcriptional levels. As the first attempt, this study proved that copepod spliced leader is useful for obtaining full-length cDNA in P. poplesia exposed to PAHs and provided a valuable gene resource for this non-model species. This approach can be applied to other calanoid copepods exposed to various stressors, particularly under field conditions. (C) 2016 Elsevier B.V. All rights reserved.
KeywordCopepod Full-length Cdna Pseudodiaptomus Poplesia Gene Expression Polycyclic Aromatic Hydrocarbons (Pahs) Spliced Leader
DOI10.1016/j.aquatox.2016.12.014
Indexed BySCI
Language英语
WOS IDWOS:000394071100013
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.qdio.ac.cn/handle/337002/136771
Collection实验海洋生物学重点实验室
Affiliation1.Ocean Univ China, Key Lab Marine Environm & Ecol, Minist Educ, Qingdao 266100, Peoples R China
2.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
3.Univ Connecticut, Dept Marine Sci, Groton, CT 06340 USA
4.State Ocean Adm, North China Sea Environm Monitoring Ctr, Qingdao 266033, Peoples R China
Recommended Citation
GB/T 7714
Zhuang, Yunyun,Yang, Feifei,Xu, Donghui,et al. Spliced leader-based analyses reveal the effects of polycyclic aromatic hydrocarbons on gene expression in the copepod Pseudodiaptomus poplesia[J]. AQUATIC TOXICOLOGY,2017,183:114-126.
APA Zhuang, Yunyun,Yang, Feifei,Xu, Donghui,Chen, Hongju,Zhang, Huan,&Liu, Guangxing.(2017).Spliced leader-based analyses reveal the effects of polycyclic aromatic hydrocarbons on gene expression in the copepod Pseudodiaptomus poplesia.AQUATIC TOXICOLOGY,183,114-126.
MLA Zhuang, Yunyun,et al."Spliced leader-based analyses reveal the effects of polycyclic aromatic hydrocarbons on gene expression in the copepod Pseudodiaptomus poplesia".AQUATIC TOXICOLOGY 183(2017):114-126.
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