壬基酚致红鲫发育畸形的机制

Preliminary studies on the mechanism of nonylphenol-induced malformation of Carassius auratus red var.

  • 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发育畸形的机制,本研究以红鲫为研究对象,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 6 121个,NNP/SCP中DEGs 7 270个。KEGG分析发现,上述DEGs多数聚集在细胞过程、环境信息处理和新陈代谢通路上。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。

     

    Abstract: Nonylphenol (NP) is the environmental endocrine disruptor that is widely present 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 object, transcriptome sequencing (RNA-seq) of normal embryos in neural stage (NC), normal embryos in 21 somites stage (SC), monstrous embryos in neural stage and monstrous embryos in 21 somite stage 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, 6 121 DEGs in NC/SC and 7 270 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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