Molecular characterization of three estrogen receptor forms in zebrafish: Binding characteristics, transactivation properties, and tissue distributions
Molecular characterization of three estrogen receptor forms in zebrafish: Binding characteristics, transactivation properties, and tissue distributions
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DOI：
10.1095/biolreprod66.6.1881
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发表时间：
2002-06-01
影响因子：
3.6
通讯作者：
Pakdel, F
中科院分区：
文献类型：
--
作者：
Menuet, A;Pellegrini, E;Pakdel, F
鱼类中有两种雌激素受体 (ER) 亚型：ERα 和 ERR，并且越来越多的证据表明 ERR 亚型有不止一种形式。然而，关于这些 ER 在成人和发育过程中的特征和功能意义的信息很少。在这里，我们报告了斑马鱼中三种功能性 ER 形式 zfERalpha、zfERbeta1 和 zfERbeta2 的克隆和表征。这些受体之间的同一性百分比表明存在三个不同的基因。每个 cDNA 编码一种特异性结合雌二醇的蛋白质，解离常数范围为 0.4 nM (zfERbeta2) 至 0.75 nM（zfERalpha 和 zfERbeta1）。在瞬时转染的细胞中，所有三种形式都能够以剂量依赖性方式诱导由共有雌激素反应元件驱动的报告基因的表达； zfERbeta2 比 zfERalpha 和 zfERbeta1 稍敏感。通过逆转录聚合酶链式反应分析的组织分布模式表明，三种 zfER mRNA 很大程度上重叠，并且主要在脑、垂体、肝脏和性腺中表达。进行原位杂交以更详细地研究成年女性大脑中三种 zfER mRNA 的分布。 zfER mRNA 在两个神经内分泌区域（视前区和下丘脑内侧基底区）表现出不同但部分重叠的表达模式。这些 zfER 的表征为理解常用于发育研究的脊椎动物物种中雌二醇作用的机制提供了新的视角。
There are two estrogen receptor (ER) subtypes in fish, ERalpha and ERR, and increasing evidence that the ERR subtype has more than one form. However, there is little information on the characteristics and functional significance of these ERs in adults and during development. Here, we report the cloning and characterization of three functional ER forms, zfERalpha, zfERbeta1, and zfERbeta2, in the zebrafish. The percentages of identity between these receptors suggest the existence of three distinct genes. Each cDNA encoded a protein that specifically bound estradiol with a dissociation constant ranging from 0.4 nM (zfERbeta2) to 0.75 nM (zfERalpha and zfERbeta1). In transiently transfected cells, all three forms were able to induce, in a dose-dependent manner, the expression of a reporter gene driven by a consensus estrogen responsive element; zfERbeta2 was slightly more sensitive than zfERalpha and zfERbeta1. Tissue distribution pattern, analyzed by reverse transcription polymerase chain reaction, showed that the three zfER mRNAs largely overlap and are predominantly expressed in brain, pituitary, liver, and gonads. In situ hybridization was performed to study in more detail the distribution of the three zfER mRNAs in the brain of adult females. The zfER mRNAs exhibit distinct but partially overlapping patterns of expression in two neuroendocrine regions, the preoptic area and the mediobasal hypothalamus. The characterization of these zfERs provides a new perspective for understanding the mechanisms underlying estradiol actions in a vertebrate species commonly used for developmental studies.