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BOR - Papers in Press, published online ahead of print January 22, 2003.
Biol Reprod 2003, 10.1095/biolreprod.102.009894
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BIOLOGY OF REPRODUCTION 68, 1982–1988 (2003)
DOI: 10.1095/biolreprod.102.009894
© 2003 by the Society for the Study of Reproduction, Inc.

Environment

Lead Reduces Messenger RNA and Protein Levels of Cytochrome P450 Aromatase and Estrogen Receptor ß in Human Ovarian Granulosa Cells1

Crystel Taupeau3, Joël Poupon5, Dominique Treton4, Aurélie Brosse3, Yolande Richard4, and Véronique Machelon2,3

INSERM Institut Paris-Sud Sur les Cytokines, Unité 355 "Maturation Gamétique et Fécondation,"3 Unité 131 "Cytokines et Immunorégulation,"4 92140 Clamart, France Laboratoire de Biochimie-Toxicologie,5 Hôpital Fernand-Widal, Paris, France

Exposure to lead causes decreased fertility in women. In the present study, we examined the in vitro effects of lead on cytochrome P450 aromatase (P450 arom) and on estrogen receptor ß (ERß), two key proteins for the human ovary. Aromatase is required for the bioconversion of androgen to estradiol; ERß mediates estrogen effects in granulosa cells. Granulosa cells were collected from women undergoing in vitro fertilization and then cultured with 10 µM lead acetate. Using atomic absorption spectrometry, we showed that lead accumulated in cells. Aromatase activity as measured by a tritiated water production assay was significantly reduced. Using semiquantitative reverse transcription-polymerase chain reaction and Western blotting procedures, we showed that P450 arom and ERß mRNA and protein content were both significantly reduced. Adding 10 µg/ml of cycloheximide, a protein inhibitor, did not eliminate the effects of lead. The present results support the hypothesis that the action of lead on fertility in women may result, in part, from the down-regulation of P450 arom and ERß gene transcription in ovarian granulosa.

1 Supported by Ministère de l'Environnement/Inserm grant EN97D06.

2 Correspondence: Véronique Machelon, INSERM Unité 355, 32 rue des Carnets, 92140 Clamart, France. FAX: 33 1 46 32 79 93; veronique.machelon{at}inserm.ipsc.u-psud.fr






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Copyright © 2003 by the Society for the Study of Reproduction.