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Dichlorodiphenyldichloroethylene Potentiates the Effect of Protein Kinase A Pathway Activators on Progesterone Synthesis in Cultured Porcine Granulosa Cells¹

^a Reproductive Biology Research Unit, Department of Obstetrics and Gynecology and Toxicology Centre, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N OW8

The insecticide dichlorodiphenyltrichloroethane (DDT) and its major metabolite p,p'-dichlorodiphenyldichloroethylene (DDE) have been implicated as endocrine-modulating chemicals. The DDT metabolite p,p'-DDE has been found contaminating human tissues and follicular fluid because of dietary exposure. We investigated the effects of DDE on progesterone synthesis in a stable porcine granulosa cell line, JC-410, and in primary cultures of porcine granulosa cells. Progesterone synthesis was not affected by 0.1–100 ng/ml DDE in the JC-410 cells. However, 10 ng/ml DDE increased 8-bromo-cAMP (8-Br-cAMP)-stimulated progesterone synthesis 0.4-fold (P < 0.05) over the levels observed with 1 mM 8-Br-cAMP alone. The effect of cholera toxin (CT) on progesterone synthesis was increased 0.7-fold (P < 0.05) by 10 ng/ml DDE over the value observed with 30 ng/ml CT alone. In primary cultures of porcine granulosa cells, 10 ng/ml DDE potentiated CT-stimulated progesterone synthesis 1.2-fold over the value observed with CT alone. In the JC-410 cells, 1 and 10 ng/ml DDE increased CT-stimulated cytochrome P450-cholesterol side-chain cleavage (P450_scc) mRNA levels 0.3- and 0.4-fold, respectively, over the values obtained with CT alone. Neither basal nor CT-stimulated cAMP levels were changed by DDE. We conclude that DDE affects granulosa cell response to protein kinase A activators by altering the expression of the P450_scc gene.

¹ This research was supported by the Canadian Network of Toxicology Centres, Saskatchewan Health (Government of Saskatchewan), Medical Research Council of Canada and Natural Sciences Engineering Research Council of Canada grants to P.J.C.

² Correspondence: P. Jorge Chedrese, Reproductive Biology Research Unit, Department of Obstetrics & Gynecology, Royal University Hospital, 103 Hospital Dr., Saskatoon, SK, Canada S7N OW8. FAX: 306 966 8040; chedresj{at}duke.usask.ca

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