In vitro effect of chlorpyrifos oxon on muscarinic receptors and adenylate cyclase.
Although the neurotoxicity of organophosphorus compounds is generally attributed to inhibition of acetylcholinesterase, recent reports have indicated that direct interactions with muscarinic receptors and signal transduction may be an additional mechanism of neurotoxicity. We have previously shown that the organophosphorus insecticide O,O-diethyl O-3,5,6-trichloro-2-pyridinyl phosphorothioate (chlorpyrifos) binds directly to muscarinic receptors and inhibits adenylate cyclase of rat striatum. We have further pursued those results in this study by investigating the effect of chlorpyrifos oxon in NG108-15 neuroblastoma-glioma cells and Chinese hamster ovary cells transfected with cDNA for human m2 or m4 muscarinic receptor subtypes. At millimolar concentrations, chlorpyrifos oxon inhibited [3H]QNB binding in all cell lines. Likewise, [3H]CD binding was inhibited in NG108-15 and CHO-Hm2 cells. When the effect of chlorpyrifos oxon on adenylate cyclase was examined, the oxon was found to inhibit adenylate cyclase at millimolar concentrations. Though this effect on cyclase required greater concentrations of oxon than the comparable effect in striatal cells, it displayed the common characteristic of being atropine-insensitive, suggesting that the effect on cyclase was not muscarinic receptor dependent. The inhibition of adenylate cyclase produced by chlorpyrifos oxon was not eliminated in pertussis toxin treated cells, lending further support to the idea that it is not a receptor-mediated event, and suggesting a potential direct interaction of chlorpyrifos oxon with the adenylate cyclase molecule.
Huff
R A
RA
Department of Pharmacology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Abou-Donia
M B
MB
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D004390:Chlorpyrifos / Q000031:analogs & derivatives* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D000242:Cyclic AMP / Q000096:biosynthesis; D005910:Glioma / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D009447:Neuroblastoma / Q000378:metabolism; D037342:Pertussis Toxin; D011976:Receptors, Muscarinic / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D010566:Virulence Factors, Bordetella / Q000633:toxicity
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D018727:Muscarinic Antagonists; D011976:Receptors, Muscarinic; D010566:Virulence Factors, Bordetella; D000246:Adenosine Diphosphate Ribose; C009618:O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000262:Adenylyl Cyclases; D004390:Chlorpyrifos