An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors

Autores
Beltrán González, Andrea Natalia; Calvo, Daniel Juan; Gasulla, Javier
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background and Purpose: Reactive oxygen species (ROS) are normally involved in cell oxidative stress but also play a role as cellular messengers in redox signalling; for example, modulating the activity of neurotransmitter receptors and ion channels. However, the direct actions of ROS on GABAA receptors were not previously demonstrated. In the present work, we studied the effects of ROS on GABAAρ1 receptor function. Experimental Approach: GABAAρ1 receptors were expressed in oocytes and GABA-evoked responses electrophysiologically recorded in the presence or absence of ROS. Chemical protection of cysteines by selective sulfhydryl reagents and site-directed mutagenesis studies were used to identify protein residues involved in ROS actions. Key Results: GABAAρ1 receptor-mediated responses were significantly enhanced in a concentration-dependent and reversible manner by H2O2. Potentiating effects were attenuated by a free radical scavenger, lipoic acid or an inhibitor of the Fenton reaction, deferoxamine. Each ρ1 subunit contains only three cysteine residues, two extracellular at the Cys-loop (C177 and C191) and one intracellular (C364) at the M3-M4 linker. Mutant GABAAρ1 receptors in which C364 was exchanged by alanine were completely insensitive to modulation, implying that this site, rather than a cysteine in the Cys-loop, is essential for ROS modulation. Conclusion and Implications: Our results show that the function of GABAAρ1 receptors is enhanced by ROS and that the intracellular C364 is the sensor for ROS actions.
Fil: Beltrán González, Andrea Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
Fil: Calvo, Daniel Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
Fil: Gasulla, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
Materia
GABAA receptors
Reactive oxygen species
Chloride channels
Retina
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/3957
Acceder
id CONICETDig_01f1bf15a8dc2b28f9ae8eedff5d75a4
oai_identifier_str oai:ri.conicet.gov.ar:11336/3957
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptorsBeltrán González, Andrea NataliaCalvo, Daniel JuanGasulla, JavierGABAA receptorsReactive oxygen speciesChloride channelsRetinahttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Background and Purpose: Reactive oxygen species (ROS) are normally involved in cell oxidative stress but also play a role as cellular messengers in redox signalling; for example, modulating the activity of neurotransmitter receptors and ion channels. However, the direct actions of ROS on GABAA receptors were not previously demonstrated. In the present work, we studied the effects of ROS on GABAAρ1 receptor function. Experimental Approach: GABAAρ1 receptors were expressed in oocytes and GABA-evoked responses electrophysiologically recorded in the presence or absence of ROS. Chemical protection of cysteines by selective sulfhydryl reagents and site-directed mutagenesis studies were used to identify protein residues involved in ROS actions. Key Results: GABAAρ1 receptor-mediated responses were significantly enhanced in a concentration-dependent and reversible manner by H2O2. Potentiating effects were attenuated by a free radical scavenger, lipoic acid or an inhibitor of the Fenton reaction, deferoxamine. Each ρ1 subunit contains only three cysteine residues, two extracellular at the Cys-loop (C177 and C191) and one intracellular (C364) at the M3-M4 linker. Mutant GABAAρ1 receptors in which C364 was exchanged by alanine were completely insensitive to modulation, implying that this site, rather than a cysteine in the Cys-loop, is essential for ROS modulation. Conclusion and Implications: Our results show that the function of GABAAρ1 receptors is enhanced by ROS and that the intracellular C364 is the sensor for ROS actions.Fil: Beltrán González, Andrea Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; ArgentinaFil: Calvo, Daniel Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; ArgentinaFil: Gasulla, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; ArgentinaWiley2014-04-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/3957Beltrán González, Andrea Natalia; Calvo, Daniel Juan; Gasulla, Javier; An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors; Wiley; British Journal of Pharmacology; 171; 9; 11-4-2014; 2291-22990007-1188enginfo:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1111/bph.12581/abstractinfo:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/DOI:10.1111/bph.12581info:eu-repo/semantics/altIdentifier/issn/0007-1188info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-03-31T15:22:59Zoai:ri.conicet.gov.ar:11336/3957instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-03-31 15:23:00.237CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
title An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
spellingShingle An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
Beltrán González, Andrea Natalia
GABAA receptors
Reactive oxygen species
Chloride channels
Retina
title_short An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
title_full An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
title_fullStr An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
title_full_unstemmed An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
title_sort An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors
dc.creator.none.fl_str_mv Beltrán González, Andrea Natalia
Calvo, Daniel Juan
Gasulla, Javier
author Beltrán González, Andrea Natalia
author_facet Beltrán González, Andrea Natalia
Calvo, Daniel Juan
Gasulla, Javier
author_role author
author2 Calvo, Daniel Juan
Gasulla, Javier
author2_role author
author
dc.subject.none.fl_str_mv GABAA receptors
Reactive oxygen species
Chloride channels
Retina
topic GABAA receptors
Reactive oxygen species
Chloride channels
Retina
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Background and Purpose: Reactive oxygen species (ROS) are normally involved in cell oxidative stress but also play a role as cellular messengers in redox signalling; for example, modulating the activity of neurotransmitter receptors and ion channels. However, the direct actions of ROS on GABAA receptors were not previously demonstrated. In the present work, we studied the effects of ROS on GABAAρ1 receptor function. Experimental Approach: GABAAρ1 receptors were expressed in oocytes and GABA-evoked responses electrophysiologically recorded in the presence or absence of ROS. Chemical protection of cysteines by selective sulfhydryl reagents and site-directed mutagenesis studies were used to identify protein residues involved in ROS actions. Key Results: GABAAρ1 receptor-mediated responses were significantly enhanced in a concentration-dependent and reversible manner by H2O2. Potentiating effects were attenuated by a free radical scavenger, lipoic acid or an inhibitor of the Fenton reaction, deferoxamine. Each ρ1 subunit contains only three cysteine residues, two extracellular at the Cys-loop (C177 and C191) and one intracellular (C364) at the M3-M4 linker. Mutant GABAAρ1 receptors in which C364 was exchanged by alanine were completely insensitive to modulation, implying that this site, rather than a cysteine in the Cys-loop, is essential for ROS modulation. Conclusion and Implications: Our results show that the function of GABAAρ1 receptors is enhanced by ROS and that the intracellular C364 is the sensor for ROS actions.
Fil: Beltrán González, Andrea Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
Fil: Calvo, Daniel Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
Fil: Gasulla, Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular; Argentina
description Background and Purpose: Reactive oxygen species (ROS) are normally involved in cell oxidative stress but also play a role as cellular messengers in redox signalling; for example, modulating the activity of neurotransmitter receptors and ion channels. However, the direct actions of ROS on GABAA receptors were not previously demonstrated. In the present work, we studied the effects of ROS on GABAAρ1 receptor function. Experimental Approach: GABAAρ1 receptors were expressed in oocytes and GABA-evoked responses electrophysiologically recorded in the presence or absence of ROS. Chemical protection of cysteines by selective sulfhydryl reagents and site-directed mutagenesis studies were used to identify protein residues involved in ROS actions. Key Results: GABAAρ1 receptor-mediated responses were significantly enhanced in a concentration-dependent and reversible manner by H2O2. Potentiating effects were attenuated by a free radical scavenger, lipoic acid or an inhibitor of the Fenton reaction, deferoxamine. Each ρ1 subunit contains only three cysteine residues, two extracellular at the Cys-loop (C177 and C191) and one intracellular (C364) at the M3-M4 linker. Mutant GABAAρ1 receptors in which C364 was exchanged by alanine were completely insensitive to modulation, implying that this site, rather than a cysteine in the Cys-loop, is essential for ROS modulation. Conclusion and Implications: Our results show that the function of GABAAρ1 receptors is enhanced by ROS and that the intracellular C364 is the sensor for ROS actions.
publishDate 2014
dc.date.none.fl_str_mv 2014-04-11
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/3957
Beltrán González, Andrea Natalia; Calvo, Daniel Juan; Gasulla, Javier; An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors; Wiley; British Journal of Pharmacology; 171; 9; 11-4-2014; 2291-2299
0007-1188
url http://hdl.handle.net/11336/3957
identifier_str_mv Beltrán González, Andrea Natalia; Calvo, Daniel Juan; Gasulla, Javier; An intracellular redox sensor for reactive oxygen species at the M3-M4 linker of GABAArho1 receptors; Wiley; British Journal of Pharmacology; 171; 9; 11-4-2014; 2291-2299
0007-1188
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1111/bph.12581/abstract
info:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/DOI:10.1111/bph.12581
info:eu-repo/semantics/altIdentifier/issn/0007-1188
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1861214684844130304
score 12.822162

Publicaciones similares