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Dichlorodiphenylchloroethylene Elevates Cytosolic Calcium Concentrations and Oscillations in Primary Cultures of Human Granulosa-Lutein Cells¹

Department of Obstetrics & Gynecology,³ Reproductive Biology Division, the Department of Medicine,⁴ McMaster University, Health Sciences Centre, Hamilton, Ontario, Canada L8N 3Z5

1,1-Dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), a metabolite of DDT (1,1-dichlorodiphenyltrichloroethane), is a persistent hormonally active environmental toxicant that has been found in human serum and follicular fluid. The objective of this study was to determine whether DDE can alter free calcium ion concentrations in the cytosol ([Ca²⁺]_cyt) of human granulosa cells. Changes in [Ca²⁺]_cyt in single cells loaded with Fura-2 were studied using a dynamic digital Ca²⁺ imaging system. At a concentration of 100 ng/ml, DDE stimulated small elevations of [Ca²⁺]_cyt accompanied by Ca²⁺ oscillations. At 1 µg DDE/ml, there was a biphasic Ca²⁺ response with marked elevations of [Ca²⁺]_cyt over time. In Ca²⁺-free medium, cells showed an initial small elevation of [Ca²⁺]_cyt, which was magnified after addition of Ca²⁺ to the medium. Washing the cells after DDE treatment failed to remove the elevated [Ca²⁺]_cyt and oscillations, both of which were eliminated by addition of EGTA. ATP also induced [Ca²⁺]_cyt elevations and oscillations, and these effects were potentiated when DDE was added. FSH induced transient [Ca²⁺]_cyt elevations, whereas hCG caused a prolonged elevation and marked oscillations in [Ca²⁺]_cyt. These results suggest that DDE at concentrations normally found in human tissues induces elevations in [Ca²⁺]_cyt in granulosa-lutein cells. Our data therefore highlight a novel mechanism through which DDE can alter endocrine homeostasis and possibly act as an endocrine toxicant.

¹ This work was supported by the Canadian Institutes of Health Research.

² Correspondence: Edward Younglai, Department of Obstetrics & Gynecology, McMaster University Medical Centre, 1200 Main St. West, Hamilton, Ontario, Canada L8N 3Z5. FAX: 905 524 2911; younglai{at}mcmaster.ca

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