Efficacy of pharmacological estrogen receptor antagonists in blocking activation of zebrafish estrogen receptors
Efficacy of pharmacological estrogen receptor antagonists in blocking activation of zebrafish estrogen receptors
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DOI：
10.1016/j.ygcen.2011.05.008
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发表时间：
2011-08-01
影响因子：
2.7
通讯作者：
Mayer, Gregory D.
中科院分区：
文献类型：
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作者：
Notch, Emily G.;Mayer, Gregory D.
各种药理激动剂、拮抗剂和选择性雌激素受体调节剂(SERM)已被用来更好地了解特定受体在各种生理过程中的作用。尽管结构和功能相似，但激动剂和拮抗剂对硬骨雌激素受体的影响知之甚少，这些研究的结果表明物种之间存在着广泛的差异。这项研究的目的是确定两个人类SERM调节三种斑马鱼雌激素受体亚型激活的能力。将斑马鱼雌激素受体1(ESR1)、雌激素受体2a(Esr2a)和雌激素受体2b(Esr2b)的全长基因克隆到表达载体中，与雌激素反应荧光素酶载体一起导入不表达雌激素受体的细胞。单独转染斑马鱼雌激素受体的细胞，然后暴露于17β-雌二醇(E-2)或17α-乙基雌二醇(EE2)中，荧光素酶活性呈剂量依赖性增加。没有一种药理拮抗剂。ICI 182,780。甲基-哌啶基-吡唑(MPP)或吡唑并[1.5-a]嘧啶(PHTPP)能够独立地反式激活任何一种斑马鱼雌激素受体的荧光素酶表达。在这三种ER拮抗剂中，只有ICI 182、780能够阻断EE2诱导的荧光素酶活性，尽管ICI 182比ICI 182多10到100倍。780是所有受体所必需的。MPP和PHTPP均不能阻断EE2诱导的与斑马鱼雌激素受体亚型结合的荧光素酶活性。这些结果表明，人类ER和斑马鱼ER配体结合的差异不够保守，不足以使SERM、MPP或PHTPP在斑马鱼中引起与人类相似的效应。(C)2011 Elsevier Inc.保留所有权利。
A variety of pharmacological agonists, antagonists and selective estrogen receptor modulators (SERM) have been used to better understand the role of specific receptors in various physiological processes. Despite similar structure and function, less is known about the effect of agonists and antagonists on teleost estrogen receptors and the results of these studies have indicated wide variation among species. The goal of this study was to determine the ability of two human SERMs to modulate activation of three zebrafish estrogen receptor isoforms. Full length cDNA of zebrafish estrogen receptor 1 (esr1), estrogen receptor 2a (esr2a) and estrogen receptor 2b (esr2b) were cloned into expression vectors and transfected into cells that do not endogenously express any estrogen receptor along with an estrogen responsive luciferase vector. Cells transfected with any of the zebrafish estrogen receptors individually and then exposed to 17 beta-estradiol (E-2) or 17 alpha-ethinylestradiol (EE2) exhibited a dose dependent increase in luciferase activity. None of the pharmacological antagonists. ICI 182, 780. methyl-piperidino-pyrazole (MPP) or pyrazolo [1.5-a] pyrimidine (PHTPP), were able to independently transactivate luciferase expression with any of the zebrafish estrogen receptors. Of the three ER antagonists, only ICI 182, 780 was able to block EE2 induced luciferase activity, although a 10 to 100-fold excess of ICI 182. 780 was necessary with all receptors. Neither MPP nor PHTPP were able to block EE2 induced luciferase activity with any isoform of zebrafish estrogen receptor. These results indicate that the difference between human ER and zebrafish ER ligand binding is not conserved enough for the SERMs MPP or PHTPP to elicit similar effects in zebrafish as those manifested in humans. (C) 2011 Elsevier Inc. All rights reserved.