title: Effects of chlorpyrifos on high-affinity choline uptake and [3H]hemicholinium-3 binding in rat brain.
abstract: High, subcutaneous doses of the organophosphorus insecticide chlorpyrifos (CPF) in adult male rats can be well-tolerated despite extensive and persistent acetylcholinesterase (AChE) inhibition. We propose that changes in acetylcholine synthesis could modulate the toxicity associated with extensive AChE inhibition following CPF exposure. High-affinity choline uptake (HACU, the rate-limiting step in acetlcholine synthesis) and binding to [3H]-hemicholinium-3 (HC-3, a specific ligand for the choline transporter) were chosen as indicators of acetylcholine synthesis. Female, Sprague-Dawley rats (220-280 g) were treated with either vehicle (peanut oil, 2 ml/kg, sc) or CPF (280 mg/kg, 2 ml/kg, sc), examined daily for clinical signs of toxicity, and sacrificed 1, 2, or 7 days later for neurochemical measurements (AChE inhibition, muscarinic receptor binding using [3H]quinuclidinyl benzilate (QNB) and [3H]cis-methyldioxolane (CD) as ligands, HACU and [3H]HC-3 binding) in frontal cortex. Despite extensive AChE inhibition (90-93%) at all time points, relatively minor degrees of overt toxicity were noted in CPF-treated rats. Binding to the non-selective muscarinic antagonist [3H]QNB was reduced (10-34%), whereas binding to the putative m2-selective agonist [3H]CD was increased (15-23%) at all three time points. HACU was reduced (20%) in crude synaptosomes prepared from CPF-treated rats 1 day following exposure but no significant changes were noted at 2 or 7 days after treatment. CPF-oxon, the active oxidative metabolite of CPF, was a weak inhibitor of HACU in vitro (IC50 > 200 microM). Binding to [3H]HC-3 was reduced in a dose-related manner 1 day after CPF exposure. Kinetic analyses of [3H]HC-3 binding 1 day after CPF (280 mg/kg) indicated a significant reduction in density (Bmax: control, 187 +/- 18 fmol/mg protein; CPF, 104 +/- 12 fmol/mg protein) with no apparent change in binding affinity (Kd: control, 25 +/- 3 nM; CPF, 19 +/- 3 nM). These results suggest that a reduction in HACU/acetylcholine synthesis may contribute, along with compensatory changes in cholinergic receptors, to the diminished toxicity following extensive AChE inhibition by CPF.
authors.affiliation: Division of Toxicology, College of Pharmacy and Health Sciences, Northeast Louisiana University, Monroe 71209-0470, USA.
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.forename: C N
authors.initials: CN
authors.lastname: Pope
mesh_terms: D000818:Animals; D004390:Chlorpyrifos / Q000494:pharmacology*; D002794:Choline / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D005260:Female; D006426:Hemicholinium 3 / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism; D014316:Tritium
chemical_list: D002800:Cholinesterase Inhibitors; D011976:Receptors, Muscarinic; D014316:Tritium; D006426:Hemicholinium 3; D004390:Chlorpyrifos; D002794:Choline