兴国红鲤ER基因克隆表达及EE2暴露对肝中ER表达的影响  

CLONE AND EXPRESSION OF ER GENES IN XINGGUO RED CARP AND THEIR EXPRESSION IN LIVER UPON EE2 EXPOSURE IN JUVENILES

作　　者：宋明月[1,2] 单喜双[1,2] 陈细华[1] 岳华梅[1] 叶欢[1] 杨晓鸽[1] 李创举[1] 

机构地区：[1]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,武汉430223 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《水生生物学报》2017年第1期26-32,共7页Acta Hydrobiologica Sinica

基　　金：国家高技术研究发展计划项目(2011AA100401)资助~~

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。The full-length cDNAs of estrogen receptors, ERα, ERβ, ERβ1 and ERβ2, were cloned from Xingguo red carp （Cyprinus carpio var singuonensis, Cs）. The amino acid sequence of Cs ERs shared the highest identity with their counterparts of three cyprinid species. The ERs highly expressed in the liver, ovary and intestines of female Cs. In male Cs, ERαand ERβmRNAs were mainly expressed in liver, and ERβ1 and ERβ2 mainly observed in intestine and testis, respectively. The mRNA level of ERαmRNAs was significantly up-regulated in in liver of Juveniles （150 dph） ex-posed to different concentrations （0.01, 0.1 and 1 nmol/L） of 17α-ethinylestradiol （EE2） for 1 and 2 weeks. The hepatic ERβmRNA expression was dramatically increased by EE2 exposure, and ERβ1 mRNA expression was suppressed by EE2 for 1 and 2 weeks. The expression of ERβ2 mRNA was suppressed in the 1-week group with the highest suppres-sion at 0.01 nmol/L of EE2. This study showed that EE2 differently regulated the expression of four ERs genes in liver. Compared to ERβ, ERβ1 and ERβ2, ERαis a more sensitive biomarker for short-term （1—2 week） EE2 exposure.

关 键 词：雌激素受体 兴国红鲤 EE2 实时荧光定量PCR 

分 类 号：Q344.1[生物学—遗传学]