Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation

Autores
Chrestia, Juan Facundo; Bruzzone, Ariana; Esandi, María del Carmen; Bouzat, Cecilia Beatriz
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The α7 receptor is a nicotinic receptor present in neuronal and non-neuronal cells. α7 acts as a ligand-gated ion channel and as a metabotropic receptor. We investigated the role of tyrosine phosphorylation of the intracellular domain (ICD) in the dual ionotropic/metabotropic receptor function. Single-channel recordings from HEK cells expressing α7 showed that channel activity appears as brief isolated openings and episodes of few openings in quick succession (bursts). Exposure to an inhibitor of Src family kinases (PP2) increased the frequency and duration of bursts while preincubation with an inhibitor of tyrosine phosphatases had the opposite effect. Co-expression of α7 and an inactive Src kinase also increased burst duration. A mutant α7 lacking tyrosine phosphorylation sites in the ICD showed longer burst durations and insensitivity to PP2, thus recapitulating the effects of phosphorylation inhibition on wild-type α7. Cells exposed to the specific α7 agonist (PNU-282987) showed an increase of ERK1/2 phosphorylation, which was abolished by exposure to a tyrosine kinase inhibitor. PNU-282987 did not trigger ERK phosphorylation in cells expressing the mutant receptor lacking tyrosine residues or co-expressing α7 and α7-ICD domain. Our results indicate that dephosphorylation positively modulates ionotropic α7 activity in a way compatible with decreased desensitization, and that the phosphorylated state of α7-ICD plays a role in metabotropic receptor responses.
Fil: Chrestia, Juan Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bruzzone, Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Esandi, María del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
XXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN)
Villa Carlos Paz
Argentina
Sociedad Argentina de Investigación en Neurociencias
Materia
ionotopic receptor
metabotropic receptor
Nicotinic receptor
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:11336/192237
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylationChrestia, Juan FacundoBruzzone, ArianaEsandi, María del CarmenBouzat, Cecilia Beatrizionotopic receptormetabotropic receptorNicotinic receptorhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The α7 receptor is a nicotinic receptor present in neuronal and non-neuronal cells. α7 acts as a ligand-gated ion channel and as a metabotropic receptor. We investigated the role of tyrosine phosphorylation of the intracellular domain (ICD) in the dual ionotropic/metabotropic receptor function. Single-channel recordings from HEK cells expressing α7 showed that channel activity appears as brief isolated openings and episodes of few openings in quick succession (bursts). Exposure to an inhibitor of Src family kinases (PP2) increased the frequency and duration of bursts while preincubation with an inhibitor of tyrosine phosphatases had the opposite effect. Co-expression of α7 and an inactive Src kinase also increased burst duration. A mutant α7 lacking tyrosine phosphorylation sites in the ICD showed longer burst durations and insensitivity to PP2, thus recapitulating the effects of phosphorylation inhibition on wild-type α7. Cells exposed to the specific α7 agonist (PNU-282987) showed an increase of ERK1/2 phosphorylation, which was abolished by exposure to a tyrosine kinase inhibitor. PNU-282987 did not trigger ERK phosphorylation in cells expressing the mutant receptor lacking tyrosine residues or co-expressing α7 and α7-ICD domain. Our results indicate that dephosphorylation positively modulates ionotropic α7 activity in a way compatible with decreased desensitization, and that the phosphorylated state of α7-ICD plays a role in metabotropic receptor responses.Fil: Chrestia, Juan Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Bruzzone, Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Esandi, María del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaXXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN)Villa Carlos PazArgentinaSociedad Argentina de Investigación en NeurocienciasSociedad Argentina de Investigación en Neurociencias2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/192237Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation; XXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN); Villa Carlos Paz; Argentina; 2019; 160-160CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://saneurociencias.org.ar/congresos-san-2/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:59:26Zoai:ri.conicet.gov.ar:11336/192237instacron: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:34982025-09-29 09:59:27.237CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
title Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
spellingShingle Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
Chrestia, Juan Facundo
ionotopic receptor
metabotropic receptor
Nicotinic receptor
title_short Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
title_full Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
title_fullStr Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
title_full_unstemmed Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
title_sort Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation
dc.creator.none.fl_str_mv Chrestia, Juan Facundo
Bruzzone, Ariana
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author Chrestia, Juan Facundo
author_facet Chrestia, Juan Facundo
Bruzzone, Ariana
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author_role author
author2 Bruzzone, Ariana
Esandi, María del Carmen
Bouzat, Cecilia Beatriz
author2_role author
author
author
dc.subject.none.fl_str_mv ionotopic receptor
metabotropic receptor
Nicotinic receptor
topic ionotopic receptor
metabotropic receptor
Nicotinic receptor
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The α7 receptor is a nicotinic receptor present in neuronal and non-neuronal cells. α7 acts as a ligand-gated ion channel and as a metabotropic receptor. We investigated the role of tyrosine phosphorylation of the intracellular domain (ICD) in the dual ionotropic/metabotropic receptor function. Single-channel recordings from HEK cells expressing α7 showed that channel activity appears as brief isolated openings and episodes of few openings in quick succession (bursts). Exposure to an inhibitor of Src family kinases (PP2) increased the frequency and duration of bursts while preincubation with an inhibitor of tyrosine phosphatases had the opposite effect. Co-expression of α7 and an inactive Src kinase also increased burst duration. A mutant α7 lacking tyrosine phosphorylation sites in the ICD showed longer burst durations and insensitivity to PP2, thus recapitulating the effects of phosphorylation inhibition on wild-type α7. Cells exposed to the specific α7 agonist (PNU-282987) showed an increase of ERK1/2 phosphorylation, which was abolished by exposure to a tyrosine kinase inhibitor. PNU-282987 did not trigger ERK phosphorylation in cells expressing the mutant receptor lacking tyrosine residues or co-expressing α7 and α7-ICD domain. Our results indicate that dephosphorylation positively modulates ionotropic α7 activity in a way compatible with decreased desensitization, and that the phosphorylated state of α7-ICD plays a role in metabotropic receptor responses.
Fil: Chrestia, Juan Facundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bruzzone, Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Esandi, María del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
XXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN)
Villa Carlos Paz
Argentina
Sociedad Argentina de Investigación en Neurociencias
description The α7 receptor is a nicotinic receptor present in neuronal and non-neuronal cells. α7 acts as a ligand-gated ion channel and as a metabotropic receptor. We investigated the role of tyrosine phosphorylation of the intracellular domain (ICD) in the dual ionotropic/metabotropic receptor function. Single-channel recordings from HEK cells expressing α7 showed that channel activity appears as brief isolated openings and episodes of few openings in quick succession (bursts). Exposure to an inhibitor of Src family kinases (PP2) increased the frequency and duration of bursts while preincubation with an inhibitor of tyrosine phosphatases had the opposite effect. Co-expression of α7 and an inactive Src kinase also increased burst duration. A mutant α7 lacking tyrosine phosphorylation sites in the ICD showed longer burst durations and insensitivity to PP2, thus recapitulating the effects of phosphorylation inhibition on wild-type α7. Cells exposed to the specific α7 agonist (PNU-282987) showed an increase of ERK1/2 phosphorylation, which was abolished by exposure to a tyrosine kinase inhibitor. PNU-282987 did not trigger ERK phosphorylation in cells expressing the mutant receptor lacking tyrosine residues or co-expressing α7 and α7-ICD domain. Our results indicate that dephosphorylation positively modulates ionotropic α7 activity in a way compatible with decreased desensitization, and that the phosphorylated state of α7-ICD plays a role in metabotropic receptor responses.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/192237
Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation; XXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN); Villa Carlos Paz; Argentina; 2019; 160-160
CONICET Digital
CONICET
url http://hdl.handle.net/11336/192237
identifier_str_mv Intracellular modulation of alpha7 ionotropic and metabotropic functions by tyrosine phosphorylation; XXXIV Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias (SAN); Villa Carlos Paz; Argentina; 2019; 160-160
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://saneurociencias.org.ar/congresos-san-2/
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
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
publisher.none.fl_str_mv Sociedad Argentina de Investigación en Neurociencias
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
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score 13.070432

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