文章摘要
壬基酚致红鲫鱼发育畸形的机制初步研究
Preliminary studies on the mechanism of nonylphenol-induced malformation of Carassius auratus red var.
投稿时间:2019-09-30  修订日期:2020-01-01
DOI:
中文关键词: 壬基酚  红鲫鱼胚胎  转录组测序  荧光定量PCR
英文关键词: Nonylphenol  Carassius auratus red var.  Transcriptome sequencing  qRT-PCR
基金项目:国家重点研发计划(2017YFD0200400);湖南省教育厅重点项目(17A072); 农业部热带亚热带水产资源利用与养殖重点实验室开放课题基金
作者单位E-mail
田雨苏 湖南科技大学生命科学学院 461073961@qq.com 
孙远东 湖南科技大学生命科学学院  
欧 密 中国水产科学研究院珠江水产研究所  
崔小娟 湖南科技大学生命科学学院  
周定港 湖南科技大学生命科学学院  
车文安 湖南科技大学生命科学学院  
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中文摘要:
      壬基酚(nonylphenol,NP)是广泛存在于水体中的环境内分泌干扰物,对生物体的生长发育造成影响。为了探究壬基酚致红鲫鱼发育畸形的机制,本研究以红鲫鱼(Carassius auratus red var.)为研究对象,采用Illumina HiSep 2500对正常神经胚期胚胎(NC)、正常21体节期胚胎(SC)和暴露5 μmol/L NP中发育畸形的神经胚期胚胎(NNP)、畸形21体节期胚胎(SNP)进行转录组测序(RNA-seq),并结合实时荧光定量PCR(qRT-PCR)验证差异表达基因(differential expression gene, DEG)。RNA-seq共得到89,166条高质量Unigenes,其中30,319条Unigenes获得注释结果。NC、SC、NNP和SNP两两比较分析发现:NC/NNP中DEGs 153个,SC/SNP中DEGs 10个,NC/SC中DEGs 6121个,NNP/SCP中DEGs 7270个。KEGG分析发现上述DEGs大多数聚集在细胞过程(Cellular Processes),环境信息处理(Environmental information Processing)和新陈代谢(Metabolism)通路上。利用qRT-PCR验证了生长、发育、细胞骨架及心血管循环相关的25个DEGs,它们的表达变化在qRT-PCR和RNA-seq中结果一致,一方面说明了RNA-seq结果可靠,另一方面初步挖掘出NP胁迫致红鲫鱼胚胎发育畸形的候选基因,为后续深入探讨NP致畸的分子机制研究提供前期数据。
英文摘要:
      Nonylphenol (NP) is the environmental endocrine disruptor that widely presents in water, which affects the growth and development of species. In order to explore the mechanism of nonylphenol-induced developmental malformation in red crucian carp (Carassius auratus red var.), red crucian carp was used as the research model, transcriptome sequencing (RNA-seq) of normal embryos in neural stage (NC), normal embryos in 21 somite stage (SC), monstrous embryos in neural stage and monstrous embryos in 21 somite stage that exposed to 5 μmol/L NP (SNP) were performed on Illumina HiSep 2500 platform. Furthermore, differentially expressed genes (DEGs) were verified by quantitative real-time PCR (qRT-PCR). A total of 89,166 high-quality Unigenes were obtained from RNA-seq, of which 30,319 Unigenes were annotated. Pairwise comparison among NC, SC, NNP and SNP showed that there were 153 DEGs in NC/NNP, 10 DEGs in SC/SNP, 6121 DEGs in NC/SC, and 7270 DEGs in NNP/SCP. KEGG pathway analysis revealed that most of these above DEGs were enriched to cellular processes, environmental information processing and metabolism signal pathways. 25 DEGs related to growth, development, cytoskeleton and cardiovascular circulation were verified by qRT-PCR. Their expression changes are consistent in qRT-PCR and RNA-seq. On the one hand, this consistency indicates that the RNA-seq results are reliable, on the other hand, the candidate genes of developmental malformation caused by NP stress are preliminarily excavated, which provides prophase data for further study on the molecular mechanism of NP teratogenesis.
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