ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury

In the spinal cord high extracellular glutamate evokes excitotoxic damage with neuronal loss and severe locomotor impairment. During the cell dysfunction process, extracellular pH becomes acid and may activate acid-sensing ion channels (ASICs) which could be important contributors to neurodegenerati...

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Autores principales:	González Inchauspe, Carlota María Fabiola, Uchitel, Osvaldo Daniel
Publicado:	2017
Materias:	acid sensing ion channels (ASICs) fictive locomotion kainic acid neuroprotection pH spinal cord injury 4',6 diamidino 2 phenylindole acid sensing ion channel acid sensing ion channel 1a acid sensing ion channel 1b acid sensing ion channel 2 acid sensing ion channel 3 unclassified drug acid sensing ion channel blocking agent glutamic acid indole derivative messenger RNA proton animal cell animal experiment animal model animal tissue Article cell counting cell death cell viability assay concentration response controlled study excitotoxicity gene expression immunohistochemistry mouse nerve cell nerve cell network nervous system electrophysiology neurotoxicity newborn nonhuman oscillatory potential pH measurement priority journal protein expression reverse transcription polymerase chain reaction staining animal cell survival disease model dose response drug effects genetics glia metabolism pathology physiology spinal cord synaptic transmission tissue culture technique Acid Sensing Ion Channel Blockers Acid Sensing Ion Channels Animals Cell Death Cell Survival Disease Models, Animal Dose-Response Relationship, Drug Glutamic Acid Indoles Kainic Acid Mice Neuroglia Neurons Protons RNA, Messenger Spinal Cord Spinal Cord Injuries Synaptic Transmission Tissue Culture Techniques
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03064522_v343_n_p398_Mazzone http://hdl.handle.net/20.500.12110/paper_03064522_v343_n_p398_Mazzone
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_03064522_v343_n_p398_Mazzone2023-06-08T15:31:19Z ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury González Inchauspe, Carlota María Fabiola Uchitel, Osvaldo Daniel acid sensing ion channels (ASICs) fictive locomotion kainic acid neuroprotection pH spinal cord injury 4',6 diamidino 2 phenylindole acid sensing ion channel acid sensing ion channel 1a acid sensing ion channel 1b acid sensing ion channel 2 acid sensing ion channel 3 kainic acid unclassified drug acid sensing ion channel acid sensing ion channel blocking agent glutamic acid indole derivative kainic acid messenger RNA proton animal cell animal experiment animal model animal tissue Article cell counting cell death cell viability assay concentration response controlled study excitotoxicity gene expression immunohistochemistry mouse nerve cell nerve cell network nervous system electrophysiology neuroprotection neurotoxicity newborn nonhuman oscillatory potential pH measurement priority journal protein expression reverse transcription polymerase chain reaction spinal cord injury staining animal cell death cell survival disease model dose response drug effects genetics glia metabolism nerve cell pathology physiology spinal cord spinal cord injury synaptic transmission tissue culture technique Acid Sensing Ion Channel Blockers Acid Sensing Ion Channels Animals Cell Death Cell Survival Disease Models, Animal Dose-Response Relationship, Drug Glutamic Acid Indoles Kainic Acid Mice Neuroglia Neurons Protons RNA, Messenger Spinal Cord Spinal Cord Injuries Synaptic Transmission Tissue Culture Techniques In the spinal cord high extracellular glutamate evokes excitotoxic damage with neuronal loss and severe locomotor impairment. During the cell dysfunction process, extracellular pH becomes acid and may activate acid-sensing ion channels (ASICs) which could be important contributors to neurodegenerative pathologies. Our previous studies have shown that transient application of the glutamate analog kainate (KA) evokes delayed excitotoxic death of spinal neurons, while white matter is mainly spared. The present goal was to enquire if ASIC channels modulated KA damage in relation to locomotor network function and cell death. Mouse spinal cord slices were treated with KA (0.01 or 0.1 mM) for 1 h, and then washed out for 24 h prior to analysis. RT-PCR results showed that KA (at 0.01 mM concentration that is near-threshold for damage) increased mRNA expression of ASIC1a, ASIC1b, ASIC2 and ASIC3, an effect reversed by the ASIC inhibitor 4′,6-diamidino-2-phenylindole (DAPI). A KA neurotoxic dose (0.1 mM) reduced ASIC1a and ASIC2 expression. Cell viability assays demonstrated KA-induced large damage in spinal slices from mice with ASIC1a gene ablation. Likewise, immunohistochemistry indicated significant neuronal loss when KA was followed by the ASIC inhibitors DAPI or amiloride. Electrophysiological recording from ventral roots of isolated spinal cords showed that alternating oscillatory cycles were slowed down by 0.01 mM KA, and intensely inhibited by subsequently applied DAPI or amiloride. Our data suggest that early rise in ASIC expression and function counteracted deleterious effects on spinal networks by raising the excitotoxicity threshold, a result with potential implications for improving neuroprotection. © 2016 IBRO Fil:Gonzalez-Inchauspe, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Uchitel, O.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2017 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03064522_v343_n_p398_Mazzone http://hdl.handle.net/20.500.12110/paper_03064522_v343_n_p398_Mazzone
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	acid sensing ion channels (ASICs) fictive locomotion kainic acid neuroprotection pH spinal cord injury 4',6 diamidino 2 phenylindole acid sensing ion channel acid sensing ion channel 1a acid sensing ion channel 1b acid sensing ion channel 2 acid sensing ion channel 3 kainic acid unclassified drug acid sensing ion channel acid sensing ion channel blocking agent glutamic acid indole derivative kainic acid messenger RNA proton animal cell animal experiment animal model animal tissue Article cell counting cell death cell viability assay concentration response controlled study excitotoxicity gene expression immunohistochemistry mouse nerve cell nerve cell network nervous system electrophysiology neuroprotection neurotoxicity newborn nonhuman oscillatory potential pH measurement priority journal protein expression reverse transcription polymerase chain reaction spinal cord injury staining animal cell death cell survival disease model dose response drug effects genetics glia metabolism nerve cell pathology physiology spinal cord spinal cord injury synaptic transmission tissue culture technique Acid Sensing Ion Channel Blockers Acid Sensing Ion Channels Animals Cell Death Cell Survival Disease Models, Animal Dose-Response Relationship, Drug Glutamic Acid Indoles Kainic Acid Mice Neuroglia Neurons Protons RNA, Messenger Spinal Cord Spinal Cord Injuries Synaptic Transmission Tissue Culture Techniques
spellingShingle	acid sensing ion channels (ASICs) fictive locomotion kainic acid neuroprotection pH spinal cord injury 4',6 diamidino 2 phenylindole acid sensing ion channel acid sensing ion channel 1a acid sensing ion channel 1b acid sensing ion channel 2 acid sensing ion channel 3 kainic acid unclassified drug acid sensing ion channel acid sensing ion channel blocking agent glutamic acid indole derivative kainic acid messenger RNA proton animal cell animal experiment animal model animal tissue Article cell counting cell death cell viability assay concentration response controlled study excitotoxicity gene expression immunohistochemistry mouse nerve cell nerve cell network nervous system electrophysiology neuroprotection neurotoxicity newborn nonhuman oscillatory potential pH measurement priority journal protein expression reverse transcription polymerase chain reaction spinal cord injury staining animal cell death cell survival disease model dose response drug effects genetics glia metabolism nerve cell pathology physiology spinal cord spinal cord injury synaptic transmission tissue culture technique Acid Sensing Ion Channel Blockers Acid Sensing Ion Channels Animals Cell Death Cell Survival Disease Models, Animal Dose-Response Relationship, Drug Glutamic Acid Indoles Kainic Acid Mice Neuroglia Neurons Protons RNA, Messenger Spinal Cord Spinal Cord Injuries Synaptic Transmission Tissue Culture Techniques González Inchauspe, Carlota María Fabiola Uchitel, Osvaldo Daniel ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
topic_facet	acid sensing ion channels (ASICs) fictive locomotion kainic acid neuroprotection pH spinal cord injury 4',6 diamidino 2 phenylindole acid sensing ion channel acid sensing ion channel 1a acid sensing ion channel 1b acid sensing ion channel 2 acid sensing ion channel 3 kainic acid unclassified drug acid sensing ion channel acid sensing ion channel blocking agent glutamic acid indole derivative kainic acid messenger RNA proton animal cell animal experiment animal model animal tissue Article cell counting cell death cell viability assay concentration response controlled study excitotoxicity gene expression immunohistochemistry mouse nerve cell nerve cell network nervous system electrophysiology neuroprotection neurotoxicity newborn nonhuman oscillatory potential pH measurement priority journal protein expression reverse transcription polymerase chain reaction spinal cord injury staining animal cell death cell survival disease model dose response drug effects genetics glia metabolism nerve cell pathology physiology spinal cord spinal cord injury synaptic transmission tissue culture technique Acid Sensing Ion Channel Blockers Acid Sensing Ion Channels Animals Cell Death Cell Survival Disease Models, Animal Dose-Response Relationship, Drug Glutamic Acid Indoles Kainic Acid Mice Neuroglia Neurons Protons RNA, Messenger Spinal Cord Spinal Cord Injuries Synaptic Transmission Tissue Culture Techniques
description	In the spinal cord high extracellular glutamate evokes excitotoxic damage with neuronal loss and severe locomotor impairment. During the cell dysfunction process, extracellular pH becomes acid and may activate acid-sensing ion channels (ASICs) which could be important contributors to neurodegenerative pathologies. Our previous studies have shown that transient application of the glutamate analog kainate (KA) evokes delayed excitotoxic death of spinal neurons, while white matter is mainly spared. The present goal was to enquire if ASIC channels modulated KA damage in relation to locomotor network function and cell death. Mouse spinal cord slices were treated with KA (0.01 or 0.1 mM) for 1 h, and then washed out for 24 h prior to analysis. RT-PCR results showed that KA (at 0.01 mM concentration that is near-threshold for damage) increased mRNA expression of ASIC1a, ASIC1b, ASIC2 and ASIC3, an effect reversed by the ASIC inhibitor 4′,6-diamidino-2-phenylindole (DAPI). A KA neurotoxic dose (0.1 mM) reduced ASIC1a and ASIC2 expression. Cell viability assays demonstrated KA-induced large damage in spinal slices from mice with ASIC1a gene ablation. Likewise, immunohistochemistry indicated significant neuronal loss when KA was followed by the ASIC inhibitors DAPI or amiloride. Electrophysiological recording from ventral roots of isolated spinal cords showed that alternating oscillatory cycles were slowed down by 0.01 mM KA, and intensely inhibited by subsequently applied DAPI or amiloride. Our data suggest that early rise in ASIC expression and function counteracted deleterious effects on spinal networks by raising the excitotoxicity threshold, a result with potential implications for improving neuroprotection. © 2016 IBRO
author	González Inchauspe, Carlota María Fabiola Uchitel, Osvaldo Daniel
author_facet	González Inchauspe, Carlota María Fabiola Uchitel, Osvaldo Daniel
author_sort	González Inchauspe, Carlota María Fabiola
title	ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
title_short	ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
title_full	ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
title_fullStr	ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
title_full_unstemmed	ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
title_sort	asic channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury
publishDate	2017
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03064522_v343_n_p398_Mazzone http://hdl.handle.net/20.500.12110/paper_03064522_v343_n_p398_Mazzone
work_keys_str_mv	AT gonzalezinchauspecarlotamariafabiola asicchannelinhibitionenhancesexcitotoxicneuronaldeathinaninvitromodelofspinalcordinjury AT uchitelosvaldodaniel asicchannelinhibitionenhancesexcitotoxicneuronaldeathinaninvitromodelofspinalcordinjury
_version_	1768545600721125376

ASIC channel inhibition enhances excitotoxic neuronal death in an in vitro model of spinal cord injury

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