title: Organophosphorus pesticides decrease M2 muscarinic receptor function in guinea pig airway nerves via indirect mechanisms.
abstract: BACKGROUND
  Epidemiological studies link organophosphorus pesticide (OP) exposures to asthma, and we have shown that the OPs chlorpyrifos, diazinon and parathion cause airway hyperreactivity in guinea pigs 24 hr after a single subcutaneous injection. OP-induced airway hyperreactivity involves M2 muscarinic receptor dysfunction on airway nerves independent of acetylcholinesterase (AChE) inhibition, but how OPs inhibit neuronal M2 receptors in airways is not known. In the central nervous system, OPs interact directly with neurons to alter muscarinic receptor function or expression; therefore, in this study we tested whether the OP parathion or its oxon metabolite, paraoxon, might decrease M2 receptor function on peripheral neurons via similar direct mechanisms.
  
  
  RESULTS
  Intravenous administration of paraoxon, but not parathion, caused acute frequency-dependent potentiation of vagally-induced bronchoconstriction and increased electrical field stimulation (EFS)-induced contractions in isolated trachea independent of AChE inhibition. However, paraoxon had no effect on vagally-induced bradycardia in intact guinea pigs or EFS-induced contractions in isolated ileum, suggesting mechanisms other than pharmacologic antagonism of M2 receptors. Paraoxon did not alter M2 receptor expression in cultured cells at the mRNA or protein level as determined by quantitative RT-PCR and radio-ligand binding assays, respectively. Additionally, a biotin-labeled fluorophosphonate, which was used as a probe to identify molecular targets phosphorylated by OPs, did not phosphorylate proteins in guinea pig cardiac membranes that were recognized by M2 receptor antibodies.
  
  
  CONCLUSIONS
  These data indicate that neither direct pharmacologic antagonism nor downregulated expression of M2 receptors contributes to OP inhibition of M2 function in airway nerves, adding to the growing evidence of non-cholinergic mechanisms of OP neurotoxicity.
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Oregon Health & Science University, Portland, Oregon, United States of America.
authors.forename: Becky J
authors.initials: BJ
authors.lastname: Proskocil
authors.forename: Donald A
authors.initials: DA
authors.lastname: Bruun
authors.forename: Charles M
authors.initials: CM
authors.lastname: Thompson
authors.forename: Allison D
authors.initials: AD
authors.lastname: Fryer
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Lein
mesh_terms: D000818:Animals; D001919:Bradycardia / Q000139:chemically induced / Q000473:pathology / Q000503:physiopathology; D016084:Bronchoconstriction / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006168:Guinea Pigs; D007082:Ileum / Q000187:drug effects / Q000294:innervation; D066298:In Vitro Techniques; D007275:Injections, Intravenous; D009119:Muscle Contraction / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010261:Paraoxon / Q000008:administration & dosage / Q000633:toxicity*; D010278:Parathion / Q000008:administration & dosage / Q000633:toxicity*; D010575:Pesticides / Q000633:toxicity*; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D043585:Receptor, Muscarinic M2 / Q000235:genetics / Q000378:metabolism*; D014132:Trachea / Q000187:drug effects* / Q000294:innervation* / Q000503:physiopathology
chemical_list: D010575:Pesticides; D012333:RNA, Messenger; D043585:Receptor, Muscarinic M2; D010278:Parathion; D010261:Paraoxon