Locomotor and Sensorimotor Performance Deficit in Rats following Exposure to Pyridostigmine Bromide, DEET, and Permethrin, Alone and in Combination

Abstract

Since their return from Persian Gulf War (PGW), many veterans have complained of symptoms including muscle and joint pain, ataxia, chronic fatigue, headache, and difficulty with concentration. The causes of the symptoms remain unknown. Because these veterans were exposed to a combination of chemicals including pyridostigmine bromide (PB), DEET, and permethrin, we investigated the effects of these agents, alone and in combination, on the sensorimotor behavior and central cholinergic system of rats. Male Sprague-Dawley rats (200-250 gm) were treated with DEET (40 mg/kg, dermal) or permethrin (0.13 mg/kg, dermal), alone and in combination with PB (1.3 mg/kg, oral, last 15 days only), for 45 days. Sensorimotor ability was assessed by a battery of behavioral tests that included beam-walk score, beam-walk time, incline plane performance, and forepaw grip on days 30 and 45 following the treatment. On day 45 the animals were sacrificed, and plasma and CNS cholinesterase, and brain choline acetyl transferase, muscarinic and nicotinic acetylcholine receptors were evaluated. Animals treated with PB, alone or in combination with DEET and permethrin, showed a significant deficit in beam-walk score as well as beam-walk time as compared with controls. Treatment with either DEET or permethrin, alone or in combination with each other, did not have a significant effect on beam-walk score. All chemicals, alone or in combination, resulted in a significant impairment in incline plane testing on days 30 and 45 following treatment. Treatment with PB, DEET, or permethrin alone did not have any inhibitory effect on plasma or brain cholinesterase activities, except that PB alone caused moderate inhibition in midbrain acetylcholinesterase (AChE) activity. Treatment with permethrin alone caused significant increase in cortical and cerebellar AChE activity. A combination of DEET and permethrin or PB and DEET led to significant decrease in AChE activity in brainstem and midbrain and brainstem, respectively. A significant decrease in brainstem AChE activity was observed following combined exposure to PB and permethrin. Coexposure with PB, DEET, and permethrin resulted in significant inhibition in AChE in brainstem and midbrain. No effect was observed on choline acetyl transferase activity in brainstem or cortex, except combined exposure to PB, DEET, and permethrin caused a slight but significant increase in cortical choline acetyltransferase activity. Treatment with PB, DEET, and permethrin alone caused a significant increase in ligand binding for m2 muscarinic acetylcholine receptor (mAChR) in the cortex. Coexposure to PB, DEET, and permethrin did not have any effect over that of PB-induced increase in ligand binding. There was no significant change in ligand binding for nicotinic acetylcholine receptor (nAChR) associated with treatment with the chemical alone; a combination of PB and DEET or coexposure with PB, DEET, and permethrin caused a significant increase in nAChR ligand binding in the cortex. Thus, these results suggest that exposure ...

Protein binding of isofluorophate in vivo after coexposure to multiple chemicals.

“…The compounds, even in total, remain at tracer concentrations (~100 nM maximum whole-body average for PYB) that can cause only minor competitive inhibition of proteins. Abou-Donia et al see no significant change in brain enzyme activities for AChE, choline acetyltransferase, m2 muscarinic acetylcholine (ACh) receptor, and nicotinic ACh receptor at PYB doses 25 times greater than those used here, despite significant changes in a measure of locomotor skills most affected by PYB alone (34). Our observed 10-15% reduction in brain DFP binding does not fit a dose-dependent extrapolation of those higher-dose studies.…”

“…This PYB dose is only 4% of the prophylactic dose used by service people in the Persian Gulf War to protect against nerve agent exposure. That dose was chosen to temporarily shield about 30% of nerve esterase from OP binding (34). The 10% protection observed here is a factor of 10 greater than a linear extrapolation of that effect.…”

Neuroscience and accelerator mass spectrometry

“…The protective nature of PYB in the brain did not fit a dose-dependent extrapolation from higher dose studies. 35 Induction of a protective esterase activity in another tissue that lowered plasma DFP, and consequently brain DFP, is consistent with our observations, as is decreased DFP bioavailability due to increased intestinal peristalsis. The induced enzyme had to be specific to DFP and not affect PTN and PER, however, since pre-exposure to these increased DFP binding.…”

Dose Dependent Prophylactic Efficacy of 6-Chlorotacrine in Soman-Poisoned Mice

“…Pyridostigmine is positively charged and, therefore, it does not readily cross BBB to afford the protection of brain AChE. In addition, a recent review emphasizes that this type of classic pharmacological pretreatment can produce behavioral impairment and region-specific alterations in ACh receptors at the doses required to afford protection against convulsant doses of nerve agents (26)(27).…”