赖静萍, 闫月明, 温茹淑, 方展强. 食蚊鱼CYP19b基因克隆及环境激素暴露对其表达的影响[J]. 水生生物学报, 2016, 40(3): 451-458. DOI: 10.7541/2016.60
引用本文: 赖静萍, 闫月明, 温茹淑, 方展强. 食蚊鱼CYP19b基因克隆及环境激素暴露对其表达的影响[J]. 水生生物学报, 2016, 40(3): 451-458. DOI: 10.7541/2016.60
LAI Jing-Ping, YAN Yue-Ming, WEN Ru-Shu, FANG Zhan-Qiang. MOLECULAR CLONING AND MRNA EXPRESSION OF CYP19B GENE INDUCED BY ENVIRONMENTAL HORMONES IN THE MOSQUITOFISH (GAMBUSIA AFFINIS)[J]. ACTA HYDROBIOLOGICA SINICA, 2016, 40(3): 451-458. DOI: 10.7541/2016.60
Citation: LAI Jing-Ping, YAN Yue-Ming, WEN Ru-Shu, FANG Zhan-Qiang. MOLECULAR CLONING AND MRNA EXPRESSION OF CYP19B GENE INDUCED BY ENVIRONMENTAL HORMONES IN THE MOSQUITOFISH (GAMBUSIA AFFINIS)[J]. ACTA HYDROBIOLOGICA SINICA, 2016, 40(3): 451-458. DOI: 10.7541/2016.60

食蚊鱼CYP19b基因克隆及环境激素暴露对其表达的影响

MOLECULAR CLONING AND MRNA EXPRESSION OF CYP19B GENE INDUCED BY ENVIRONMENTAL HORMONES IN THE MOSQUITOFISH (GAMBUSIA AFFINIS)

  • 摘要: 为探讨17-雌二醇(E2)、17-甲基睾酮(MT)和雄烯二酮(AED)暴露对食蚊鱼脑组织CYP19b表达的影响,首次克隆和分析了食蚊鱼(Gambusia affinis) CYP19b cDNA的全系列。采用静水暴露方法, 分别设置2、10、50和100 ng/L E2、MT和AED各4个实验组, 并设置对照组和平行组, 定量测定了1、3、5和7d脑组织中的CYP19b mRNA表达水平的变化。成功克隆了食蚊鱼CYP19b基因全长, 获得基因片段总长1670 bp, ORF(OpenReading Frame)是从52 bp到1554 bp, 共编码501个氨基酸。通过与其他相关鱼类CYP19b基因编码的氨基酸序列相似性作比较, 食蚊鱼CYP19b基因氨基酸序列的同源性很高。结果显示, 短时间的暴露, E2较低浓度组(2和10 ng/L)对食蚊鱼作用不明显, 但E2相对较高浓度组(50和100 ng/L)能显著地上调食蚊鱼CYP19b基因的表达(P0.01)。暴露早期MT对CYP19b表达无明显影响, 随着暴露时间的持续延长, 除了MT较高浓度组(50 ng/L)外, 其他各浓度组MT对CYP19b的表达有显著的抑制作用(P0.01), 并呈一定的剂量相关关系。AED作为雄激素类似物与MT的作用效果相似, 相对较高浓度(50和100 ng/L)的AED抑制CYP19b的表达(P0.01)。研究结果初步表明, 3种典型环境激素对食蚊鱼CYP19b基因表达的影响十分显著, 但不同的暴露浓度及不同的取样时间点可能表现为促进或抑制CYP19b基因的表达。

     

    Abstract: The current study investiaged effects of 17-estradiol (E2), 17-methyl testosterone (MT) and androgen analogues androstendione (AED) exposure on brain tissue CYP19b gene expression in Gambusia affinis. The G. affinis CYP19b cDNA of full sequence was cloned and analyzed for the first time. G. affinis were randomly divided into five groups, with one control group and four experimental groups (2, 10, 50 and 100 ng/L E2, MT and AED, respectively). The CYP19b mRNA expression levels in fish brain tissues were determined after 1, 3, 5, and 7d exposure, respectively. The G. affinis CYP19b cDNA of full sequence was cloned, from which 1670 bp nucleotides were obtained, encoding 501 amino acids with an open reading frame (ORF) from 52 bp to 1554 bp. The comparison among the similarity amino acid sequence of CYP19b with other related fishes supports high similarity between species. The results indicated that E2 at high concentrations (50 ng/L and 100 ng/L groups) significantly up-regulated CYP19b gene expression (P0.01) but E2 at low concentration (2 ng/L and 10 ng/L groups) did not. MT had no significant effect on CYP19b expression in early stages, but it significantly inhibited CYP19b expression with prolonged treatment except the relatively high concentrations of MT (50 ng/L group) with the characerisitc of dose correlativity. As androgen analogue, AED was found to have similar effects with MT, and the relatively high concentrations (50 ng/L and 100 ng/L groups) of AED inhibited the CYP19b expression (P0.01). The results indicated the pivotal role of three typical environmental hormones under different concentration and various time points on the expression of CYP19b.

     

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