Shi Wen-Jun, Zhao Jian-Liang, Jiang Yu-Xia, Huang Guo-Yong, Liu You-Sheng, Zhang Jin-Na, Ying Guang-Guo
State Key Laboratory of Organic Geochemistry, CAS Research Centre of PRD Environmental Pollution and Control, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou, China.
University of Chinese Academy of Sciences, Beijing, China.
Environ Toxicol Chem. 2017 Dec;36(12):3267-3276. doi: 10.1002/etc.3894. Epub 2017 Aug 7.
The aim of the present study was to investigate the effects of norgestrel (NGT) on gonadal development in adult zebrafish. Adult zebrafish were exposed to NGT for 14 d at 871 ng L for microarray analysis, and a follow-up experiment was conducted to further study the targeted pathway in adult zebrafish after exposure to NGT at 6.7, 83, and 912 ng L by quantitative polymerase chain reaction (qPCR) and histological analysis. The microarray analysis revealed that 11 545 transcripts were identified. Gene ontology analysis showed organ development, system development, multicellular organismal development, single-organism developmental process, and developmental process were significantly enriched. A Venn diagram displayed 434 target genes involved in organ development, and these genes were common in these 5 development-related processes. Kyoto Encyclopedia of Genes and Genomes analysis showed that the notch signaling pathway was the top toxicity pathway, and it was selected as the target pathway for further qPCR analysis. The qPCR analysis revealed significant and dose-dependent alterations of most target genes involved in the notch signaling pathway in the gonads, even at an environmentally relevant concentration of 6.7 ng L . The transcriptional patterns were consistent with the notch signaling cascade. In addition, NGT significantly increased the frequency of mature sperm and decreased the frequency of immature sperm at all concentrations. Meanwhile, NGT treatment increased the percentage of mature vitellogenic oocytes and atretic follicles at 912 ng L but decreased the percentage of immature vitellogenic oocytes. Thus, the present study demonstrated significant developmental toxicity in the gonad of adult zebrafish even at environmentally relevant NGT concentrations. Environ Toxicol Chem 2017;36:3267-3276. © 2017 SETAC.
本研究的目的是调查炔诺孕酮（NGT）对成年斑马鱼性腺发育的影响。成年斑马鱼在871 ng/L的浓度下暴露于NGT 14天以进行微阵列分析，并进行后续实验，通过定量聚合酶链反应（qPCR）和组织学分析，进一步研究成年斑马鱼在6.7、83和912 ng/L的浓度下暴露于NGT后的靶向通路。微阵列分析显示共鉴定出11545个转录本。基因本体分析表明，器官发育、系统发育、多细胞生物体发育、单细胞生物体发育过程以及发育过程均显著富集。维恩图显示有434个参与器官发育的靶基因，这些基因在这5个与发育相关的过程中是共有的。京都基因与基因组百科全书分析表明，Notch信号通路是首要毒性通路，并被选为进一步qPCR分析的靶标通路。qPCR分析显示，即使在环境相关浓度6.7 ng/L下，性腺中大多数参与Notch信号通路的靶基因也有显著的剂量依赖性变化。转录模式与Notch信号级联一致。此外，在所有浓度下，NGT均显著增加了成熟精子的频率，降低了未成熟精子的频率。同时，在912 ng/L时，NGT处理增加了成熟卵黄生成卵母细胞和闭锁卵泡的百分比，但降低了未成熟卵黄生成卵母细胞的百分比。因此，本研究表明，即使在与环境相关的NGT浓度下，成年斑马鱼的性腺也存在显著的发育毒性。《环境毒理学与化学》2017年；36:3267 - 3276。© 2017 SETAC。
