A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala
We established a novel brain slice assay to test the ability of acetylcholinesterase (AChE) reactivators to prevent ACh-induced M1 muscarinic acetylcholine receptor (mAChR) dependent hyperexcitability observed after exposure to the organophosphate (OP)-based AChE inhibitor and sarin surrogate 4-nitr...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2022.1066312/full |
| _version_ | 1811319213794525184 |
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| author | Jeffrey S. Thinschmidt Scott W. Harden Michael A. King James D. Talton Charles J. Frazier |
| author_facet | Jeffrey S. Thinschmidt Scott W. Harden Michael A. King James D. Talton Charles J. Frazier |
| author_sort | Jeffrey S. Thinschmidt |
| collection | DOAJ |
| description | We established a novel brain slice assay to test the ability of acetylcholinesterase (AChE) reactivators to prevent ACh-induced M1 muscarinic acetylcholine receptor (mAChR) dependent hyperexcitability observed after exposure to the organophosphate (OP)-based AChE inhibitor and sarin surrogate 4-nitrophenyl isopropyl methylphosphonate (NIMP). Whole-cell patch clamp recordings were used to evaluate the response of pyramidal neurons in the rat basolateral amygdala (BLA) to brief (1 min) bath application of ACh (100 μM), either in control conditions, or after exposure to NIMP ± an AChE reactivator. Bath application of ACh produced atropine- and pirenzepine-sensitive inward currents in voltage clamped BLA pyramidal neurons, and increased the frequency of spontaneous EPSCs, suggesting robust activation of M1 mAChRs. Responses to ACh were increased ~3–5 fold in slices that had been preincubated in NIMP, and these effects were reversed in a concentration dependent manner by exposure to a commercially available AChE reactivator. The current work outlines a simple assay that can be used to evaluate the efficacy of both known and novel AChE reactivators in an area of the limbic system that likely contributes to seizures after acute exposure to OP-based AChE inhibitors. |
| first_indexed | 2024-04-13T12:37:54Z |
| format | Article |
| id | doaj.art-3545c7d60f87475388b0638bb2d7565a |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-04-13T12:37:54Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-3545c7d60f87475388b0638bb2d7565a2022-12-22T02:46:35ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022022-11-011610.3389/fncel.2022.10663121066312A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdalaJeffrey S. Thinschmidt0Scott W. Harden1Michael A. King2James D. Talton3Charles J. Frazier4Department of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville, FL, United StatesDepartment of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville, FL, United StatesAlchem Laboratories Corporation, Alachua, FL, United StatesAlchem Laboratories Corporation, Alachua, FL, United StatesDepartment of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville, FL, United StatesWe established a novel brain slice assay to test the ability of acetylcholinesterase (AChE) reactivators to prevent ACh-induced M1 muscarinic acetylcholine receptor (mAChR) dependent hyperexcitability observed after exposure to the organophosphate (OP)-based AChE inhibitor and sarin surrogate 4-nitrophenyl isopropyl methylphosphonate (NIMP). Whole-cell patch clamp recordings were used to evaluate the response of pyramidal neurons in the rat basolateral amygdala (BLA) to brief (1 min) bath application of ACh (100 μM), either in control conditions, or after exposure to NIMP ± an AChE reactivator. Bath application of ACh produced atropine- and pirenzepine-sensitive inward currents in voltage clamped BLA pyramidal neurons, and increased the frequency of spontaneous EPSCs, suggesting robust activation of M1 mAChRs. Responses to ACh were increased ~3–5 fold in slices that had been preincubated in NIMP, and these effects were reversed in a concentration dependent manner by exposure to a commercially available AChE reactivator. The current work outlines a simple assay that can be used to evaluate the efficacy of both known and novel AChE reactivators in an area of the limbic system that likely contributes to seizures after acute exposure to OP-based AChE inhibitors.https://www.frontiersin.org/articles/10.3389/fncel.2022.1066312/fullorganophosphatesbasolateral amygdalaacetylcholinesterasestatus epilepticusNIMPHI-6 |
| spellingShingle | Jeffrey S. Thinschmidt Scott W. Harden Michael A. King James D. Talton Charles J. Frazier A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala Frontiers in Cellular Neuroscience organophosphates basolateral amygdala acetylcholinesterase status epilepticus NIMP HI-6 |
| title | A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| title_full | A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| title_fullStr | A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| title_full_unstemmed | A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| title_short | A rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| title_sort | rapid in vitro assay for evaluating the effects of acetylcholinesterase inhibitors and reactivators in the rat basolateral amygdala |
| topic | organophosphates basolateral amygdala acetylcholinesterase status epilepticus NIMP HI-6 |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2022.1066312/full |
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