Chlorpyrifos intermittent exposure enhances cardiovascular but not behavioural responses to contextual fear conditioning in adult rats: Possible involvement of brain oxidative-nitrosative stress

Abstract

Exposure to organophosphorus compounds (OPs) may cause psychiatric, neurologic, biochemical, and cardiovascular abnormalities. Neurotoxicity of OP compounds is primarily due to irreversibly inhibition of the acetylcholinesterase (AChE) enzyme both centrally and peripherally. Chlorpyrifos (CPF) is a widely used OP classified as moderately toxic. Previously, it has been shown that CPF administration, given every other day to adult rats, impairs spatial memory and prepulse inhibition associated with brain AChE inhibition. Our group also found that intermittent treatment with CPF, simulating occupational exposure, impairs the cardiorespiratory reflexes and causes cardiac hypertrophy. Thereby, we aimed to examine whether subchronic and intermittent administration of CPF would affect the behavioural (freezing) and cardiovascular (mean arterial pressure, MAP; heart rate, HR) responses elicited during contextual fear conditioning (CFC) and extinction. Wistar adult male rats were injected with sublethal and intermittent CPF doses (4 and 7 mg/kg) three times a week for one month. Two days after the last injection, a range of tests were performed to assess depression (sucrose preference), anxiety (elevated plus-maze, EPM), locomotion (open field, OF), and conditioned fear expression and extinction. Separate cohorts of animals were euthanized to measure plasma butyrylcholinesterase (BChE), erythrocyte AChE, brain AChE activity, and markers of oxidative-nitrosative stress. Intermittent CPF treatment did not affect sucrose preference. CPF (4 and 7 mg/kg) reduced open-arms exploration in the EPM, suggesting an anxiogenic effect. The higher dose of CPF decreased the total distance travelled in the OFT, suggesting motor impairment. After a seven-day CPF-free washout period, CPF (7 mg/kg) increased the tachycardic response without affecting freezing behaviour in the CFC extinction session. CPF 7 mg/kg decreased AChE activity in the hippocampus, pre-frontal cortex and brainstem 72 after the last administration whilst transiently increasing oxidative-nitrosative stress specifically in the brainstem. Overall, our results outlined the behavioural, autonomic and biochemical abnormalities caused by an intermittent dosing regimen of CPF that elicits brain AChE inhibition and brain oxidative-nitrosative stress. This paradigm might be valuable in further exploring long-term consequences and mechanisms of OP neurotoxicity as well as comprehensive therapeutic approaches.

Keywords: Acetylcholinesterase; Anxiety; Cardiovascular; Fear; Organophosphorus; Oxidative stress.