Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors

Autores
Nielsen, Beatriz Elizabeth; Minguez, Teresa; Bermudez, Isabel; Bouzat, Cecilia Beatriz
Año de publicación
2017
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The α7 nicotinic receptor (nAChR) is a promising drug target for neurological and inflammatory disorders. It has been considered the homomeric member of the family. The recent discovery of α7β2 receptor in brain led to the urgent need of its functional characterization. Our main goal is to determine the stoichiometry of the heteromeric α7β2 receptor and its activation and potentiation profile. We generated receptors with fixed stoichiometry by two different approaches. One involved the generation of concatemeric α7β2 pentamers of different stoichiometries, and the other involved co-expression of unlinked α7 and β2 subunits, with the α7 subunit carrying a reporter mutation. Receptors were expressed in mammalian cells and function was evaluated by single-channel recordings. We found that α7 can assemble with one, two or three β2 subunits to form functional receptors. As the number of β2 subunits in the pentamer increases, the durations of openings and activation episodes, called bursts, increase progressively whereas channel conductance remains constant. We proposed that the prolonged bursts observed for α7β2 can be used as the signature of the presence of heteromeric receptors in native tissues. The prolonged activation episodes and reduced desensitization of α7β2 may have an important impact on calcium-dependent intracellular signaling and neuronal excitability. By using mutant subunits, we demonstrated that activation of α7β2 occurs through the α7/α7 binding-site interface. Among α7 positive allosteric modulators (PAMs), which emerge as novel therapeutic tools, type I PAMs were more selective for α7 than for α7β2 whereas PNU-120596, a type II PAM, similarly potentiated all α7-containing receptors. Statistically significance differences were established at p-values<0.05.This first single-channel study of α7β2 provides basis for deciphering the role and functional location of this novel receptor and opens doors for the development of selective therapeutic drugs
Fil: Nielsen, Beatriz Elizabeth. 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: Minguez, Teresa. Oxford Brookes University (oxford Brookes University);
Fil: Bermudez, Isabel. Oxford Brookes University (oxford Brookes University);
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
LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiologia
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
Materia
ALPHA7BETA2 NICOTINIC RECEPTORS
CONCATEMERS
PATCH CLAMP
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:11336/237040
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptorsNielsen, Beatriz ElizabethMinguez, TeresaBermudez, IsabelBouzat, Cecilia BeatrizALPHA7BETA2 NICOTINIC RECEPTORSCONCATEMERSPATCH CLAMPhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The α7 nicotinic receptor (nAChR) is a promising drug target for neurological and inflammatory disorders. It has been considered the homomeric member of the family. The recent discovery of α7β2 receptor in brain led to the urgent need of its functional characterization. Our main goal is to determine the stoichiometry of the heteromeric α7β2 receptor and its activation and potentiation profile. We generated receptors with fixed stoichiometry by two different approaches. One involved the generation of concatemeric α7β2 pentamers of different stoichiometries, and the other involved co-expression of unlinked α7 and β2 subunits, with the α7 subunit carrying a reporter mutation. Receptors were expressed in mammalian cells and function was evaluated by single-channel recordings. We found that α7 can assemble with one, two or three β2 subunits to form functional receptors. As the number of β2 subunits in the pentamer increases, the durations of openings and activation episodes, called bursts, increase progressively whereas channel conductance remains constant. We proposed that the prolonged bursts observed for α7β2 can be used as the signature of the presence of heteromeric receptors in native tissues. The prolonged activation episodes and reduced desensitization of α7β2 may have an important impact on calcium-dependent intracellular signaling and neuronal excitability. By using mutant subunits, we demonstrated that activation of α7β2 occurs through the α7/α7 binding-site interface. Among α7 positive allosteric modulators (PAMs), which emerge as novel therapeutic tools, type I PAMs were more selective for α7 than for α7β2 whereas PNU-120596, a type II PAM, similarly potentiated all α7-containing receptors. Statistically significance differences were established at p-values<0.05.This first single-channel study of α7β2 provides basis for deciphering the role and functional location of this novel receptor and opens doors for the development of selective therapeutic drugsFil: Nielsen, Beatriz Elizabeth. 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: Minguez, Teresa. Oxford Brookes University (oxford Brookes University);Fil: Bermudez, Isabel. Oxford Brookes University (oxford Brookes University);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; ArgentinaLXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de ProtozoologíaBuenos AiresArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de Investigación Bioquímica y Biología MolecularSociedad Argentina de InmunologíaSociedad Argentina de AndrologíaSociedad Argentina de BiofísicaSociedad Argentina de BiologíaSociedad Argentina de Farmacología ExperimentalSociedad Argentina de FisiologiaSociedad Argentina de HematologíaSociedad Argentina de ProtozoologíaFundación Revista MedicinaKotsias, BasilioBecú Villalobos, DamasiaSemeniuk, Guillermo Basilio2017info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/237040Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 594-5950025-76801667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:06:40Zoai:ri.conicet.gov.ar:11336/237040instacron: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:34982025-09-03 10:06:41.127CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
title Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
spellingShingle Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
Nielsen, Beatriz Elizabeth
ALPHA7BETA2 NICOTINIC RECEPTORS
CONCATEMERS
PATCH CLAMP
title_short Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
title_full Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
title_fullStr Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
title_full_unstemmed Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
title_sort Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors
dc.creator.none.fl_str_mv Nielsen, Beatriz Elizabeth
Minguez, Teresa
Bermudez, Isabel
Bouzat, Cecilia Beatriz
author Nielsen, Beatriz Elizabeth
author_facet Nielsen, Beatriz Elizabeth
Minguez, Teresa
Bermudez, Isabel
Bouzat, Cecilia Beatriz
author_role author
author2 Minguez, Teresa
Bermudez, Isabel
Bouzat, Cecilia Beatriz
author2_role author
author
author
dc.contributor.none.fl_str_mv Kotsias, Basilio
Becú Villalobos, Damasia
Semeniuk, Guillermo Basilio
dc.subject.none.fl_str_mv ALPHA7BETA2 NICOTINIC RECEPTORS
CONCATEMERS
PATCH CLAMP
topic ALPHA7BETA2 NICOTINIC RECEPTORS
CONCATEMERS
PATCH CLAMP
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 nicotinic receptor (nAChR) is a promising drug target for neurological and inflammatory disorders. It has been considered the homomeric member of the family. The recent discovery of α7β2 receptor in brain led to the urgent need of its functional characterization. Our main goal is to determine the stoichiometry of the heteromeric α7β2 receptor and its activation and potentiation profile. We generated receptors with fixed stoichiometry by two different approaches. One involved the generation of concatemeric α7β2 pentamers of different stoichiometries, and the other involved co-expression of unlinked α7 and β2 subunits, with the α7 subunit carrying a reporter mutation. Receptors were expressed in mammalian cells and function was evaluated by single-channel recordings. We found that α7 can assemble with one, two or three β2 subunits to form functional receptors. As the number of β2 subunits in the pentamer increases, the durations of openings and activation episodes, called bursts, increase progressively whereas channel conductance remains constant. We proposed that the prolonged bursts observed for α7β2 can be used as the signature of the presence of heteromeric receptors in native tissues. The prolonged activation episodes and reduced desensitization of α7β2 may have an important impact on calcium-dependent intracellular signaling and neuronal excitability. By using mutant subunits, we demonstrated that activation of α7β2 occurs through the α7/α7 binding-site interface. Among α7 positive allosteric modulators (PAMs), which emerge as novel therapeutic tools, type I PAMs were more selective for α7 than for α7β2 whereas PNU-120596, a type II PAM, similarly potentiated all α7-containing receptors. Statistically significance differences were established at p-values<0.05.This first single-channel study of α7β2 provides basis for deciphering the role and functional location of this novel receptor and opens doors for the development of selective therapeutic drugs
Fil: Nielsen, Beatriz Elizabeth. 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: Minguez, Teresa. Oxford Brookes University (oxford Brookes University);
Fil: Bermudez, Isabel. Oxford Brookes University (oxford Brookes University);
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
LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología
Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Sociedad Argentina de Inmunología
Sociedad Argentina de Andrología
Sociedad Argentina de Biofísica
Sociedad Argentina de Biología
Sociedad Argentina de Farmacología Experimental
Sociedad Argentina de Fisiologia
Sociedad Argentina de Hematología
Sociedad Argentina de Protozoología
description The α7 nicotinic receptor (nAChR) is a promising drug target for neurological and inflammatory disorders. It has been considered the homomeric member of the family. The recent discovery of α7β2 receptor in brain led to the urgent need of its functional characterization. Our main goal is to determine the stoichiometry of the heteromeric α7β2 receptor and its activation and potentiation profile. We generated receptors with fixed stoichiometry by two different approaches. One involved the generation of concatemeric α7β2 pentamers of different stoichiometries, and the other involved co-expression of unlinked α7 and β2 subunits, with the α7 subunit carrying a reporter mutation. Receptors were expressed in mammalian cells and function was evaluated by single-channel recordings. We found that α7 can assemble with one, two or three β2 subunits to form functional receptors. As the number of β2 subunits in the pentamer increases, the durations of openings and activation episodes, called bursts, increase progressively whereas channel conductance remains constant. We proposed that the prolonged bursts observed for α7β2 can be used as the signature of the presence of heteromeric receptors in native tissues. The prolonged activation episodes and reduced desensitization of α7β2 may have an important impact on calcium-dependent intracellular signaling and neuronal excitability. By using mutant subunits, we demonstrated that activation of α7β2 occurs through the α7/α7 binding-site interface. Among α7 positive allosteric modulators (PAMs), which emerge as novel therapeutic tools, type I PAMs were more selective for α7 than for α7β2 whereas PNU-120596, a type II PAM, similarly potentiated all α7-containing receptors. Statistically significance differences were established at p-values<0.05.This first single-channel study of α7β2 provides basis for deciphering the role and functional location of this novel receptor and opens doors for the development of selective therapeutic drugs
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Journal
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/11336/237040
Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 594-595
0025-7680
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/237040
identifier_str_mv Stoichiometry and kinetics of activation and potentiation of nicotinic alpha7beta2 heteromeric receptors; LXII Reunión Científica de la Sociedad Argentina de Investigación Clínica; LIII Reunión Anual de la Sociedad Argentina de Investigación Bioquímica y Biología Molecular; LXV Reunión Anual de la Sociedad Argentina de Inmunología; Reunión de la Sociedad Argentina de Andrología; XLVI Reunión Anual de la Sociedad Argentina de Biofísica ; XLIX Reunión Anual de la Sociedad Argentina de Farmacología Experimental; Reunión Anual de la Sociedad Argentina de Fisiología; Reunión de la Sociedad Argentina de Hematología y XXIX Reunión Anual de la Sociedad Argentina de Protozoología; Buenos Aires; Argentina; 2017; 594-595
0025-7680
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://medicinabuenosaires.com/revistas/vol77-17/Vol.77SuplementoI-2017.pdf
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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