title: Comparative effects of oral chlorpyrifos exposure on cholinesterase activity and muscarinic receptor binding in neonatal and adult rat heart.
abstract: Organophosphorus (OP) pesticides elicit acute toxicity by inhibiting acetylcholinesterase (AChE), the enzyme responsible for inactivating acetylcholine (ACh) at cholinergic synapses. A number of OP toxicants have also been reported to interact directly with muscarinic receptors, in particular the M(2) muscarinic subtype. Parasympathetic innervation to the heart primarily regulates cardiac function by activating M(2) receptors in the sinus node, atrial-ventricular node and conducting tissues. Thus, OP insecticides can potentially influence cardiac function in a receptor-mediated manner indirectly by inhibiting acetylcholinesterase and directly by binding to muscarinic M(2) receptors. Young animals are generally more sensitive than adults to the acute toxicity of OP insecticides and age-related differences in potency of direct binding to muscarinic receptors by some OP toxicants have been reported. We thus compared the effects of the common OP insecticide chlorpyrifos (CPF) on functional signs of toxicity and cardiac cholinesterase (ChE) activity and muscarinic receptor binding in neonatal and adult rats. Dosages were based on acute lethality (i.e., 0.5 and 1x LD(10): neonates, 7.5 and 15 mg/kg; adults, 68 and 136 mg/kg). Dose- and time-related changes in body weight and cholinergic signs of toxicity (involuntary movements) were noted in both age groups. With 1x LD(10), relatively similar maximal reductions in ChE activity (95%) and muscarinic receptor binding (approximately 30%) were noted, but receptor binding reductions appeared earlier in adults and were more prolonged in neonates. In vitro inhibition studies indicated that ChE in neonatal tissues was markedly more sensitive to inhibition by the active metabolite of chlorpyrifos (i.e., chlorpyrifos oxon, CPO) than enzyme in adult tissues (IC(50) values: neonates, 17 nM; adults, 200 nM). Chelation of free calcium with EDTA had relatively little effect on in vitro cholinesterase inhibition, suggesting that differential A-esterase activity was not responsible for the age-related difference in cholinesterase sensitivity between age groups. Pre-incubation of neonatal and adult tissues with selective inhibitors of AChE and butyrylcholinesterase (BChE) indicated that a majority (82-90%) of ChE activity in the heart of both neonates and adults was BChE. The rapid onset (by 4h after dosing) of changes in muscarinic receptor binding in adult heart may be a reflection of the more potent direct binding to muscarinic receptors by chlorpyrifos oxon previously reported in adult tissues. The results suggest that ChE activity (primarily BChE) in neonatal heart may be inherently more sensitive to inhibition by some anticholinesterases and that toxicologically significant binding to muscarinic receptors may be possible with acute chlorpyrifos intoxication, potentially contributing to age-related differences in sensitivity.
authors.affiliation: Department of Physiological Sciences, College of Veterinary Medicine, 264 McElroy Hall, Oklahoma State University, Stillwater, OK 74078, United States.
authors.forename: Marcia D
authors.initials: MD
authors.lastname: Howard
authors.forename: Nikita
authors.initials: N
authors.lastname: Mirajkar
authors.forename: Subramanya
authors.initials: S
authors.lastname: Karanth
authors.forename: Carey N
authors.initials: CN
authors.lastname: Pope
mesh_terms: D000284:Administration, Oral; D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D004390:Chlorpyrifos / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000633:toxicity; D002802:Cholinesterases / Q000378:metabolism*; D005260:Female; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D020128:Inhibitory Concentration 50; D008297:Male; D018721:Muscarinic Agonists / Q000494:pharmacology; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism; D010095:Oxotremorine / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D011976:Receptors, Muscarinic / Q000378:metabolism*; D013770:Tetraisopropylpyrophosphamide / Q000494:pharmacology; D015430:Weight Gain / Q000187:drug effects; D015431:Weight Loss / Q000187:drug effects
chemical_list: D018721:Muscarinic Agonists; D011976:Receptors, Muscarinic; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; D013770:Tetraisopropylpyrophosphamide; C009618:O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; D010095:Oxotremorine; D002802:Cholinesterases; D004390:Chlorpyrifos