title: Rapid effects of pesticides on human granulosa-lutein cells.
abstract: Following our previous demonstration that p,p'-DDE (dichlorodiphenylchloroethylene), at environmentally relevant concentrations, can rapidly increase intracellular calcium [Ca2+]i concentrations in human granulosa-lutein cells, we examined whether other pesticides, such as Kepone, o,p-DDE and methoxychlor, have similar effects. Cultured human granulosa-lutein cells were loaded with Fura-2 AM, and changes in [Ca2+]i concentrations within small areas of single cells were studied with a dynamic digital Ca2+ imaging system. Kepone, at concentrations of 0.2-2 nmol/ml, consistently increased [Ca2+]i concentrations 2-6 times higher than baseline values within minutes of exposure. Methoxychlor at concentrations of 2.8-280 nmol/ml failed to alter [Ca2+]i levels consistently in cells from 10 patients. However, at 0.28 and 1.4 nmol/ml, increases in [Ca2+]i concentrations could be elicited by methoxychlor. The isomer o,p-DDE at 3 nmol/ml increased [Ca2+]i in granulosa cells of 11/20 patients. Pertussis toxin treatment inhibited the [Ca2+]i increases induced by estradiol, p,p'-DDE, o,p-DDE and methoxychlor, but not by Kepone or progesterone, indicating that Kepone and progesterone may act through an insensitive G protein-coupled receptor. The [Ca2+]i increases induced by Kepone also occurred in Ca2+-free medium, suggesting that [Ca2+]i mobilization occurred from the smooth endoplasmic reticulum. Thapsigargin and cyclopiazonic acid, two inhibitors of the endoplasmic reticulum Ca2+ pump, also stimulated [Ca2+]i increases but did not inhibit the Ca2+ response to all the pesticides. These results demonstrate that pesticides can have a rapid effect on human granulosa-lutein cells, and a nongenomic mechanism of action is suggested.
authors.affiliation: Department of Obstetrics and Gynecology, Reproductive Biology Division, McMaster University, Health Sciences Centre, 1200 Main Street West, Hamilton, Ontario, Canada, L8N 3Z5.
authors.forename: Yanjing
authors.initials: Y
authors.lastname: Wu
authors.forename: Warren G
authors.initials: WG
authors.lastname: Foster
authors.forename: Edward V
authors.initials: EV
authors.lastname: Younglai
mesh_terms: D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D007631:Chlordecone / Q000494:pharmacology*; D003600:Cytosol / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D016257:Fura-2; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008184:Luteal Cells / Q000187:drug effects* / Q000378:metabolism; D008731:Methoxychlor / Q000494:pharmacology*; D008856:Microscopy, Fluorescence; D008939:Mitotane / Q000031:analogs & derivatives / Q000494:pharmacology; D037342:Pertussis Toxin / Q000494:pharmacology; D010575:Pesticides / Q000494:pharmacology*; D011374:Progesterone / Q000494:pharmacology; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D012815:Signal Processing, Computer-Assisted; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D004791:Enzyme Inhibitors; D007211:Indoles; D010575:Pesticides; D011374:Progesterone; D019284:Thapsigargin; D008939:Mitotane; C054919:2,2-(2-chlorophenyl-4'-chlorophenyl)-1,1-dichloroethene; D037342:Pertussis Toxin; D000252:Calcium-Transporting ATPases; D007631:Chlordecone; D008731:Methoxychlor; D002118:Calcium; D016257:Fura-2; C000543:cyclopiazonic acid