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Regular Article |
a Endocrinologie Moléculaire de la Reproduction, UMR CNRS 6026, Campus de Beaulieu, 35042 Rennes cedex, France
b Laboratoire de Biologie Moléculaire et Cellulaire, UMR CNRS 5665, Ecole Normale Supérieure de Lyon, 69364 Lyon cedex 07, France
There are two estrogen receptor (ER) subtypes in fish, ER
and ERß, and increasing evidence that the ERß 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, zfER, zfERß1, and zfERß2, 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 (zfERß2) to 0.75 nM (zfER and zfERß1). 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; zfERß2 was slightly more sensitive than zfER and zfERß1. 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.
1 This work was supported by grants from the CNRS, the European Union, and the Fondation Langlois. A.M. is the recipient of a fellowship from the French Ministry of Research and Education.
2 Correspondence. FAX: 33 2 23 23 67 94; farzad.pakdel{at}univ-rennes1.fr
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