Divergent roles for glutathione in lindane-induced acute and delayed-onset inhibition of rat myometrial gap junctions.
Previous studies have shown that the insecticide lindane (gamma-hexachlorocyclohexane) induces a biphasic inhibition of gap junction intercellular communication that is accompanied by oxidative stress. The present study investigates the hypothesis that depletion of cellular glutathione (GSH) is a mechanistic link between lindane-induced oxidative stress and inhibition of myometrial gap junctions. Exposure to 100 or 200 microM lindane rapidly (within 1 min) increased myometrial cell generation of superoxide, as measured by superoxide dismutase-inhibitable cytochrome c reduction, and superoxide production remained elevated for up to 60 min of exposure. To measure gap junction communication, Lucifer yellow dye was injected into myometrial cells, and dye transfer to adjoining cells was monitored. Cells were exposed to lindane with or without GSH modulators, and dye transfer was determined at the end of a 1-h exposure to 100 microM lindane (acute phase) and 24 h after termination of lindane exposure (secondary phase). The acute phase of lindane-induced inhibition of dye transfer was prevented by GSH depletion with L-buthionine-[S,R]-sulfoximine (BSO) and enhanced by GSH augmentation with GSH monoethyl ester or L-2-oxothiazolidine-4-carboxylate (OTC). In contrast, the secondary, delayed-onset phase of lindane-induced inhibition of dye transfer was enhanced by GSH depletion with BSO and prevented by GSH augmentation with GSH monoethyl ester or OTC. Changes in cellular GSH by the pharmacological modulators were confirmed by high performance liquid chromatography. These results suggest that GSH is required in the acute phase but protects against the secondary phase of lindane-induced inhibition of myometrial gap junctions.
Loch-Caruso
Rita
R
Toxicology Program, Department of Environmental Health, University of Michigan, Ann Arbor, MI 48109-2029, USA. rlc@umich.edu
Upham
Brad L
BL
Harris
Craig
C
Trosko
James E
JE
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005456:Fluorescent Dyes; D017629:Gap Junctions / Q000187:drug effects* / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D001556:Hexachlorocyclohexane / Q000633:toxicity*; D007306:Insecticides / Q000633:toxicity*; D007546:Isoquinolines; D009215:Myometrium / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors
D005456:Fluorescent Dyes; D007306:Insecticides; D007546:Isoquinolines; D013438:Sulfhydryl Compounds; D013481:Superoxides; D001556:Hexachlorocyclohexane; C017475:lucifer yellow; D005978:Glutathione