Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro
Pinnatoxins (PnTXs) A-H constitute an emerging family belonging to the cyclic imine group of phycotoxins. Interest has been focused on these fast-acting and highly-potent toxins because they are widely found in contaminated shellfish. Despite their highly complex molecular structure, PnTXs have been...
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MDPI AG
2019-05-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/17/5/306 |
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| author | Evelyne Benoit Aurélie Couesnon Jiri Lindovsky Bogdan I. Iorga Rómulo Aráoz Denis Servent Armen Zakarian Jordi Molgó |
| author_facet | Evelyne Benoit Aurélie Couesnon Jiri Lindovsky Bogdan I. Iorga Rómulo Aráoz Denis Servent Armen Zakarian Jordi Molgó |
| author_sort | Evelyne Benoit |
| collection | DOAJ |
| description | Pinnatoxins (PnTXs) A-H constitute an emerging family belonging to the cyclic imine group of phycotoxins. Interest has been focused on these fast-acting and highly-potent toxins because they are widely found in contaminated shellfish. Despite their highly complex molecular structure, PnTXs have been chemically synthetized and demonstrated to act on various nicotinic acetylcholine receptor (nAChR) subtypes. In the present work, PnTX-A, PnTX-G and analogue, obtained by chemical synthesis with a high degree of purity (>98%), have been studied in vivo and in vitro on adult mouse and isolated nerve-muscle preparations expressing the mature muscle-type (α1)<sub>2</sub>β1δε nAChR. The results show that PnTX-A and G acted on the neuromuscular system of anesthetized mice and blocked the compound muscle action potential (CMAP) in a dose- and time-dependent manner, using a minimally invasive electrophysiological method. The CMAP block produced by both toxins in vivo was reversible within 6−8 h. PnTX-A and G, applied to isolated extensor digitorum longus nerve-muscle preparations, blocked reversibly isometric twitches evoked by nerve stimulation. The action of PnTX-A was reversed by 3,4-diaminopyridine. Both toxins exerted no direct action on muscle fibers, as revealed by direct muscle stimulation. PnTX-A and G blocked synaptic transmission at mouse neuromuscular junctions and PnTX-A amino ketone analogue (containing an open form of the imine ring) had no effect on neuromuscular transmission. These results indicate the importance of the cyclic imine for interacting with the adult mammalian muscle-type nAChR. Modeling and docking studies revealed molecular determinants responsible for the interaction of PnTXs with the muscle-type nAChR. |
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| format | Article |
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| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-11T22:32:42Z |
| publishDate | 2019-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-309c1c6ae51141cc925bfe382303791f2022-12-22T03:59:18ZengMDPI AGMarine Drugs1660-33972019-05-0117530610.3390/md17050306md17050306Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In VitroEvelyne Benoit0Aurélie Couesnon1Jiri Lindovsky2Bogdan I. Iorga3Rómulo Aráoz4Denis Servent5Armen Zakarian6Jordi Molgó7Commissariat à l’Energie Atomique et aux énergies Alternatives (CEA), Institut des Sciences du Vivant Frédéric Joliot, Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), CEA de Saclay, Université Paris-Saclay, F-91191 Gif-sur-Yvette, FranceCentre National de la Recherche Scientifique (CNRS), Institut des Neurosciences Paris-Saclay (Neuro-PSI), UMR 9197 CNRS/Université Paris-Sud, F-91198 Gif-sur-Yvette, FranceCentre National de la Recherche Scientifique (CNRS), Institut des Neurosciences Paris-Saclay (Neuro-PSI), UMR 9197 CNRS/Université Paris-Sud, F-91198 Gif-sur-Yvette, FranceCentre National de la Recherche Scientifique (CNRS), Institut de Chimie des Substances Naturelles, UPR 2301, Labex LERMIT, F-91198 Gif-sur-Yvette, FranceCommissariat à l’Energie Atomique et aux énergies Alternatives (CEA), Institut des Sciences du Vivant Frédéric Joliot, Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), CEA de Saclay, Université Paris-Saclay, F-91191 Gif-sur-Yvette, FranceCommissariat à l’Energie Atomique et aux énergies Alternatives (CEA), Institut des Sciences du Vivant Frédéric Joliot, Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), CEA de Saclay, Université Paris-Saclay, F-91191 Gif-sur-Yvette, FranceDepartment of Chemistry and Biochemistry, University of California, Santa Barbara, CA 93106, USACommissariat à l’Energie Atomique et aux énergies Alternatives (CEA), Institut des Sciences du Vivant Frédéric Joliot, Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), CEA de Saclay, Université Paris-Saclay, F-91191 Gif-sur-Yvette, FrancePinnatoxins (PnTXs) A-H constitute an emerging family belonging to the cyclic imine group of phycotoxins. Interest has been focused on these fast-acting and highly-potent toxins because they are widely found in contaminated shellfish. Despite their highly complex molecular structure, PnTXs have been chemically synthetized and demonstrated to act on various nicotinic acetylcholine receptor (nAChR) subtypes. In the present work, PnTX-A, PnTX-G and analogue, obtained by chemical synthesis with a high degree of purity (>98%), have been studied in vivo and in vitro on adult mouse and isolated nerve-muscle preparations expressing the mature muscle-type (α1)<sub>2</sub>β1δε nAChR. The results show that PnTX-A and G acted on the neuromuscular system of anesthetized mice and blocked the compound muscle action potential (CMAP) in a dose- and time-dependent manner, using a minimally invasive electrophysiological method. The CMAP block produced by both toxins in vivo was reversible within 6−8 h. PnTX-A and G, applied to isolated extensor digitorum longus nerve-muscle preparations, blocked reversibly isometric twitches evoked by nerve stimulation. The action of PnTX-A was reversed by 3,4-diaminopyridine. Both toxins exerted no direct action on muscle fibers, as revealed by direct muscle stimulation. PnTX-A and G blocked synaptic transmission at mouse neuromuscular junctions and PnTX-A amino ketone analogue (containing an open form of the imine ring) had no effect on neuromuscular transmission. These results indicate the importance of the cyclic imine for interacting with the adult mammalian muscle-type nAChR. Modeling and docking studies revealed molecular determinants responsible for the interaction of PnTXs with the muscle-type nAChR.https://www.mdpi.com/1660-3397/17/5/306pinnatoxinsmarine phycotoxinsmouse neuromuscular systemcompound muscle action potentialsynaptic potentialsemerging toxinscyclic imines |
| spellingShingle | Evelyne Benoit Aurélie Couesnon Jiri Lindovsky Bogdan I. Iorga Rómulo Aráoz Denis Servent Armen Zakarian Jordi Molgó Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro Marine Drugs pinnatoxins marine phycotoxins mouse neuromuscular system compound muscle action potential synaptic potentials emerging toxins cyclic imines |
| title | Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro |
| title_full | Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro |
| title_fullStr | Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro |
| title_full_unstemmed | Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro |
| title_short | Synthetic Pinnatoxins A and G Reversibly Block Mouse Skeletal Neuromuscular Transmission In Vivo and In Vitro |
| title_sort | synthetic pinnatoxins a and g reversibly block mouse skeletal neuromuscular transmission in vivo and in vitro |
| topic | pinnatoxins marine phycotoxins mouse neuromuscular system compound muscle action potential synaptic potentials emerging toxins cyclic imines |
| url | https://www.mdpi.com/1660-3397/17/5/306 |
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