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Effects of multiple doses of organophosphates on evoked potentials in mouse diaphragm
S S Kelly1, G E de Blaquière, F M Williams
1Department of Environmental and Occupational Medicine, Medical School, University of Newcastle upon Tyne.
Abstract:
1. Male albino mice were injected s.c. with an organophosphate (mipafox, ecothiopate or paraoxon). Treatments were either a single injection or multiple daily injections with lower doses for 5 or 8 days. At 3 h after injection the activity of brain and diaphragm acetylcholinesterase and of brain neuropathy target esterase (NTE) was measured. Also measured in the diaphragm at 3 h post dose was the duration of spontaneous miniature endplate potentials (eMEPPs), recorded extracellularly. 2. At 7 and 28 days after dosing action potentials and evoked endplate potentials, produced by stimulating the phrenic nerve at 30 Hz, were recorded in diaphragm muscle. The amplitudes, time-course and latencies of these potentials were measured and the variability of latencies (jitter) was calculated. 3. Single doses of mipafox (20 mg/kg), ecothiopate (0.192 mg/kg) or paraoxon (0.415 mg/kg) in the mouse produced ca. 70% inhibition of diaphragm acetylcholinesterase at 3 h after dosing. All three OPs produced a prolongation of the half-decay times of eMEPPs. 4. All three OPs in the above single doses produced increased muscle action potential (postjunctional) jitter but only mipafox produced an increase in endplate potential (prejunctional) jitter. Mipafox in a slightly reduced single dose (17.5 mg/kg) had no effect on prejunctional or postjunctional jitter. 5. Multiple dosing with mipafox (8 mg/kg daily for 5 days) increased both postjunctional and prejunctional jitter at both 7 and 28 days after the end of dosing. After multiple dosing with mipafox (5 mg/kg daily for 5 days) postjunctional (but not prejunctional) jitter was increased. Multiple doses of paraoxon (0.166 mg/kg daily for 5 days) or ecothiopate (0.76 mg/kg daily for 5 days) increased prejunctional and postjunctional jitter. 6. Depending on the dosing regime, all three OPs tested were capable of increasing both prejunctional and postjunctional jitter. Neither ecothiopate nor paraoxon inhibited NTE, so this prejunctional effect is not likely to be related to 'classical' OP-induced delayed neuropathy. The prejunctional effects may be related to long-term inhibition of acetylcholinesterase and the triggering mechanism for increase in prejunctional jitter may involve a relationship between the inhibition of acetylcholinesterase and the time for which it is inhibited. The differences between the time-courses of increases in prejunctional and postjunctional jitter and the differential effects of the different multiple dosing regimes indicate that it is likely that the triggering relationship between enzyme inhibition and time is different for prejunctional and postjunctional effects.
Insights
Organophosphates (OPs) can increase neuromuscular junction (NMJ) dysfunction in mice, affecting both prejunctional and postjunctional components. The study highlights that different dosing regimens of OPs differentially impact NMJ function, suggesting distinct mechanisms for pre- and postjunctional effects.
Area of Science:
- Neuroscience
- Toxicology
- Pharmacology
Background:
- Organophosphates (OPs) are widely used pesticides and nerve agents.
- OPs are known to inhibit acetylcholinesterase (AChE), an enzyme crucial for neurotransmission.
- Neuropathy Target Esterase (NTE) is a key enzyme implicated in organophosphate-induced delayed neuropathy (OPIDN).
Purpose of the Study:
- To investigate the effects of single and multiple doses of different OPs on neuromuscular junction (NMJ) function in mice.
- To differentiate between prejunctional and postjunctional effects at the NMJ following OP exposure.
- To explore the relationship between AChE inhibition, NTE activity, and NMJ dysfunction.
Main Methods:
- Male albino mice were administered single or multiple subcutaneous injections of mipafox, ecothiopate, or paraoxon.
- Acetylcholinesterase (AChE) and Neuropathy Target Esterase (NTE) activities were measured in brain and diaphragm.
- Spontaneous miniature endplate potentials (eMEPPs), action potentials, and evoked endplate potentials were recorded in the diaphragm muscle.
- Jitter (variability of latencies) was calculated for prejunctional and postjunctional components.
Main Results:
- Single doses of mipafox, ecothiopate, and paraoxon inhibited diaphragm AChE and prolonged eMEPP half-decay times.
- All tested OPs increased postjunctional jitter; mipafox also increased prejunctional jitter after single dosing.
- Multiple dosing regimens with mipafox, ecothiopate, and paraoxon differentially increased prejunctional and postjunctional jitter at 7 and 28 days post-dosing.
- Neither ecothiopate nor paraoxon inhibited NTE, suggesting prejunctional effects are not solely due to classical OPIDN.
Conclusions:
- Organophosphates can induce both prejunctional and postjunctional neuromuscular junction dysfunction in mice.
- The observed prejunctional effects may be linked to prolonged acetylcholinesterase inhibition rather than NTE inhibition.
- Different dosing strategies of OPs lead to distinct patterns of NMJ dysfunction, indicating complex, dose- and time-dependent mechanisms.