兴国红鲤ER基因克隆表达及EE2暴露对肝中ER表达的影响

发布时间：2018-12-18 03:45

【摘要】：为评估环境内分泌干扰物对鱼类的影响,研究克隆了兴国红鲤雌激素受体(Estrogen receptor,ER)4种亚型ERα、ERβ、ERβ1、ERβ2的全长c DNA序列,氨基酸序列比对发现兴国红鲤ERs分别与3种鲤科鱼类相应亚型具有较高的同源性。实时荧光定量PCR(q RT-PCR)检测结果表明,4种ER亚型m RNA在雌雄成体组织中呈现差异表达,雌性个体中,肝、卵巢和肠中4种ERs的表达量均较高;在雄性个体中,ERα和ERβ主要在肝中表达,ERβ1、ERβ2分别在肠和精巢中的表达量最高。将150日龄的兴国红鲤幼鱼分别暴露在0.01、0.1和1 nmol/L的17α-乙炔基雌二醇(EE2)中4周,检测了雌性幼鱼肝中4种ER基因的表达变化情况。在EE2中暴露1—2周后,兴国红鲤雌性幼鱼肝中ERα基因的表达水平有极显著的提升;各浓度EE2能持续显著促进其肝中ERβ的表达;在1—2周内各浓度EE2对ERβ1表达有所抑制;第1周EE2能够抑制ERβ2基因m RNA的表达,并在0.01 nmol/L时抑制作用达到了显著的水平。上述研究结果表明,EE2暴露能诱导或抑制兴国红鲤雌性幼鱼肝中ER亚型的表达,相对于ERβ、ERβ1和ERβ2、ERα可作为EE2短期(1—2周)敏感性生物学标记。
[Abstract]:In order to evaluate the effects of environmental endocrine disruptors on fish, the full-length c DNA sequences of ER 伪, ER 尾, ER 尾 1ER 尾 2 subtypes of estrogen receptor (Estrogen receptor,ER) in Xingguo red carp were cloned. Amino acid sequence alignment showed that the ERs of Xingguo Red Carp had high homology with the corresponding subtypes of three Cyprinidae fishes. The results of real-time fluorescence quantitative PCR (q RT-PCR showed that the four ER subtypes m RNA were differentially expressed in female and male adult tissues, and the four ERs expression levels were higher in female individuals, liver, ovary and intestine. In male individuals, ER 伪 and ER 尾 were mainly expressed in the liver, and ER 尾 1 and ER 尾 2 were the highest in the intestine and testis, respectively. The expression of four kinds of ER genes in the liver of female juvenile red carp were detected by exposure to 17 伪 -acetylestradiol (EE2) for 0. 01 and 1 nmol/L for 4 weeks, respectively. After being exposed to EE2 for 1-2 weeks, the expression of ER 伪 gene in the liver of female young fish of Xingguo red carp was significantly increased, the expression of ER 尾 in liver was significantly increased by EE2 at different concentrations, and the expression of ER 尾 1 was inhibited by EE2 at different concentrations within 1-2 weeks. At the first week, EE2 could inhibit the expression of m RNA of ER 尾 2 gene, and the inhibitory effect reached a significant level at 0. 01 nmol/L. The results showed that EE2 exposure could induce or inhibit the expression of ER subtypes in the liver of young female red carp Carp. Compared with ER 尾, ER 尾 1 and ER 尾 2ER 伪, they could be used as biological markers of EE2 sensitivity in the short term (1-2 weeks).
【作者单位】：中国水产科学研究院长江水产研究所农业部淡水生物多样性保护重点实验室;上海海洋大学水产与生命学院;
【基金】：国家高技术研究发展计划项目(2011AA100401)资助~~
【分类号】：S917.4

【相似文献】