Mechanisms of chlorpyrifos and diazinon induced neurotoxicity in cortical culture.
The main action of organophosphorous insecticides is generally believed to be the inhibition of acetylcholinesterase (AChE). However, these compounds also inhibit many other enzymes, any of which may play a role in their toxicity. We tested the neurotoxic mechanism of two organophosphorous insecticides, chlorpyrifos and diazinon in primary cortical cultures. Exposure to the insecticides caused a concentration-dependent toxicity that could not be directly attributed to the oxon forms of the compounds which caused little toxicity but strongly inhibited AChE. Addition of 1 mM acetylcholine or carbachol actually attenuated the toxicity of chlorpyrifos and diazinon, and the muscarinic receptor antagonist, atropine, and the nicotinic receptor antagonist, mecamylamine, did not attenuate the toxicity of either insecticide. These results strongly suggest that the organophosphorous toxicity observed in this culture system is not mediated by buildup of extracellular acetylcholine resulting from inhibition of AChE. The toxicity of chlorpyrifos was attenuated by antagonists of either the NMDA or AMPA/kainate-type glutamate receptors, but the cell death was potentiated by the caspase inhibitor ZVAD. Diazinon toxicity was not affected by glutamate receptor antagonists, but was attenuated by ZVAD. Chlorpyrifos induced diffuse nuclear staining characteristic of necrosis, while diazinon induced chromatin condensation characteristic of apoptosis. Also, chlorpyrifos exposure increased the levels of extracellular glutamate, while diazinon did not. The results suggest two different mechanisms of neurotoxicity of the insecticides, neither one of which involved acetylcholine. Chlorpyrifos induced a glutamate-mediated excitotoxicity, while diazinon induced apoptotic neuronal death.
Rush
T
T
Department of Biomedical Sciences, Marquette University, Milwaukee, WI 53233, USA.
Liu
X Q
XQ
Hjelmhaug
J
J
Lobner
D
D
D000109:Acetylcholine / Q000378:metabolism; D000110:Acetylcholinesterase / Q000378:metabolism; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis; D061945:Caspase Inhibitors; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004390:Chlorpyrifos / Q000633:toxicity*; D002800:Cholinesterase Inhibitors / Q000633:toxicity; D003976:Diazinon / Q000633:toxicity*; D005110:Extracellular Space / Q000201:enzymology; D007306:Insecticides / Q000633:toxicity*; D051379:Mice; D009336:Necrosis; D018092:Kainic Acid Receptors / Q000037:antagonists & inhibitors / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000502:physiology
D000109:Acetylcholine; D000110:Acetylcholinesterase; D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D004390:Chlorpyrifos; D002800:Cholinesterase Inhibitors; D003976:Diazinon; D007306:Insecticides; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone