title: Several synthetic progestins disrupt the glial cell specific-brain aromatase expression in developing zebra fish.
abstract: The effects of some progestins on fish reproduction have been recently reported revealing the hazard of this class of steroidal pharmaceuticals. However, their effects at the central nervous system level have been poorly studied until now. Notwithstanding, progesterone, although still widely considered primarily a sex hormone, is an important agent affecting many central nervous system functions. Herein, we investigated the effects of a large set of synthetic ligands of the nuclear progesterone receptor on the glial-specific expression of the zebrafish brain aromatase (cyp19a1b) using zebrafish mechanism-based assays. Progesterone and 24 progestins were first screened on transgenic cyp19a1b-GFP zebrafish embryos. We showed that progesterone, dydrogesterone, drospirenone and all the progesterone-derived progestins had no effect on GFP expression. Conversely, all progestins derived from 19-nortesterone induced GFP in a concentration-dependent manner with EC50 ranging from the low nM range to hundreds nM. The 19-nortestosterone derived progestins levonorgestrel (LNG) and norethindrone (NET) were further tested in a radial glial cell context using U251-MG cells co-transfected with zebrafish ER subtypes (zfERα, zfERβ1 or zfERβ2) and cyp19a1b promoter linked to luciferase. Progesterone had no effect on luciferase activity while NET and LNG induced luciferase activity that was blocked by ICI 182,780. Zebrafish-ERs competition assays showed that NET and LNG were unable to bind to ERs, suggesting that the effects of these compounds on cyp19a1b require metabolic activation prior to elicit estrogenic activity. Overall, we demonstrate that 19-nortestosterone derived progestins elicit estrogenic activity by inducing cyp19a1b expression in radial glial cells. Given the crucial role of radial glial cells and neuro-estrogens in early development of brain, the consequences of exposure of fish to these compounds require further investigation.
authors.affiliation: Team NEED, Institut de recherche en Santé Environnement et Travail (Irset), INSERM U1085, Université de Rennes 1, Campus de Beaulieu, SFR Biosit, 35042 Rennes cedex, France.
authors.forename: Joel
authors.initials: J
authors.lastname: Cano-Nicolau
authors.affiliation: Unité d'Ecotoxicologie in vitro et in vivo, Institut National de l'Environnement Industriel et des Risques (INERIS), BP 2, 60550 Verneuil-en-Halatte, France.
authors.forename: Clémentine
authors.initials: C
authors.lastname: Garoche
authors.affiliation: Unité d'Ecotoxicologie in vitro et in vivo, Institut National de l'Environnement Industriel et des Risques (INERIS), BP 2, 60550 Verneuil-en-Halatte, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Hinfray
authors.affiliation: Team NEED, Institut de recherche en Santé Environnement et Travail (Irset), INSERM U1085, Université de Rennes 1, Campus de Beaulieu, SFR Biosit, 35042 Rennes cedex, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Pellegrini
authors.affiliation: TREK, Institut de recherche en Santé Environnement et Travail (Irset), INSERM U1085, Université de Rennes 1, Campus de Beaulieu, SFR Biosit, 35042 Rennes cedex, France.
authors.forename: Noureddine
authors.initials: N
authors.lastname: Boujrad
authors.affiliation: TREK, Institut de recherche en Santé Environnement et Travail (Irset), INSERM U1085, Université de Rennes 1, Campus de Beaulieu, SFR Biosit, 35042 Rennes cedex, France.
authors.forename: Farzad
authors.initials: F
authors.lastname: Pakdel
authors.affiliation: Team NEED, Institut de recherche en Santé Environnement et Travail (Irset), INSERM U1085, Université de Rennes 1, Campus de Beaulieu, SFR Biosit, 35042 Rennes cedex, France. Electronic address: oliver.kah@univ-rennes1.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Kah
authors.affiliation: Unité d'Ecotoxicologie in vitro et in vivo, Institut National de l'Environnement Industriel et des Risques (INERIS), BP 2, 60550 Verneuil-en-Halatte, France. Electronic address: francois.brion@ineris.fr.
authors.forename: François
authors.initials: F
authors.lastname: Brion
mesh_terms: D000728:Androgens / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D004958:Estradiol / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009457:Neuroglia / Q000187:drug effects* / Q000378:metabolism; D045167:Progesterone Congeners / Q000494:pharmacology*; D011960:Receptors, Estrogen / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000728:Androgens; D004967:Estrogens; D045167:Progesterone Congeners; D011960:Receptors, Estrogen; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins; D013739:Testosterone; D004958:Estradiol; D001141:Aromatase
keywords: Brain aromatase; Progestins; Radial glial cells; Zebrafish; cyp19a1b-GFP