Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle

Detalhes bibliográficos
Autor(a) principal: Duarte, Thiago [UNIFESP]
Data de Publicação: 2012
Outros Autores: Menezes-Rodrigues, Francisco Sandro [UNIFESP], Godinho, Rosely Oliveira [UNIFESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://dx.doi.org/10.1124/jpet.112.192997
http://repositorio.unifesp.br/handle/11600/34918
Resumo: beta(2)-Adrenoceptor (beta(2)-AR) agonists increase skeletal muscle contractile force via activation of G(s) protein/adenylyl cyclases (AC) and increased generation of cAMP. Herein, we evaluated the possible dual coupling of beta(2)-AR to G(s) and G(i) proteins and the influence of the beta(2)-AR/G(s)-G(i) /cAMP signaling cascade on skeletal muscle contraction. Assuming that the increment of intracellular cAMP is followed by cAMP efflux and extracellular generation of adenosine, the contribution of the extracellular cAMP-adenosine pathway on the beta(2)-AR inotropic response was also addressed. the effects of clenbuterol/fenoterol (beta(2)-AR agonists), forskolin (AC activator), cAMP/8-bromo-cAMP, and adenosine were evaluated on isometric contractility of mouse diaphragm muscle induced by supramaximal direct electrical stimulation (0.1 Hz, 2 ms duration). Clenbuterol/fenoterol (10-1000 mu M), 1 mu M forskolin, and 20 mu M rolipram induced transient positive inotropic effects that peaked 30 min after stimulation onset, declining to 10 to 20% of peak levels in 30 min. the late descending phase of the beta(2)-AR agonist inotropic effect was mimicked by either cAMP or adenosine and abolished by preincubation of diaphragm with pertussis toxin (PTX) (G(i) signaling inhibitor) or the organic anion transporter inhibitor probenecid, indicating a delayed coupling of beta(2)-AR to G(i) protein which depends on cAMP efflux. Remarkably, the PTX-sensitive beta(2)-AR inotropic effect was inhibited by the A(1) adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine and ecto-5'-phosphodiesterase inhibitor alpha,beta-methyleneadenosine 5'-diphosphate sodium salt, indicating that beta(2)-AR coupling to G(i) is indirect and dependent on A(1) receptor activation. the involvement of the extracellular cAMP-adenosine pathway in beta(2)-AR signaling would provide a negative feedback loop that may limit stimulatory G protein-coupled receptor positive inotropism and potential deleterious effects of excessive contractile response.
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spelling Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Musclebeta(2)-Adrenoceptor (beta(2)-AR) agonists increase skeletal muscle contractile force via activation of G(s) protein/adenylyl cyclases (AC) and increased generation of cAMP. Herein, we evaluated the possible dual coupling of beta(2)-AR to G(s) and G(i) proteins and the influence of the beta(2)-AR/G(s)-G(i) /cAMP signaling cascade on skeletal muscle contraction. Assuming that the increment of intracellular cAMP is followed by cAMP efflux and extracellular generation of adenosine, the contribution of the extracellular cAMP-adenosine pathway on the beta(2)-AR inotropic response was also addressed. the effects of clenbuterol/fenoterol (beta(2)-AR agonists), forskolin (AC activator), cAMP/8-bromo-cAMP, and adenosine were evaluated on isometric contractility of mouse diaphragm muscle induced by supramaximal direct electrical stimulation (0.1 Hz, 2 ms duration). Clenbuterol/fenoterol (10-1000 mu M), 1 mu M forskolin, and 20 mu M rolipram induced transient positive inotropic effects that peaked 30 min after stimulation onset, declining to 10 to 20% of peak levels in 30 min. the late descending phase of the beta(2)-AR agonist inotropic effect was mimicked by either cAMP or adenosine and abolished by preincubation of diaphragm with pertussis toxin (PTX) (G(i) signaling inhibitor) or the organic anion transporter inhibitor probenecid, indicating a delayed coupling of beta(2)-AR to G(i) protein which depends on cAMP efflux. Remarkably, the PTX-sensitive beta(2)-AR inotropic effect was inhibited by the A(1) adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine and ecto-5'-phosphodiesterase inhibitor alpha,beta-methyleneadenosine 5'-diphosphate sodium salt, indicating that beta(2)-AR coupling to G(i) is indirect and dependent on A(1) receptor activation. the involvement of the extracellular cAMP-adenosine pathway in beta(2)-AR signaling would provide a negative feedback loop that may limit stimulatory G protein-coupled receptor positive inotropism and potential deleterious effects of excessive contractile response.Universidade Federal de São Paulo, Escola Paulista Med, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Pharmacol, Div Cellular Pharmacol, BR-04044020 São Paulo, BrazilWeb of ScienceFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Amer Soc Pharmacology Experimental TherapeuticsUniversidade Federal de São Paulo (UNIFESP)Duarte, Thiago [UNIFESP]Menezes-Rodrigues, Francisco Sandro [UNIFESP]Godinho, Rosely Oliveira [UNIFESP]2016-01-24T14:27:17Z2016-01-24T14:27:17Z2012-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion820-828http://dx.doi.org/10.1124/jpet.112.192997Journal of Pharmacology and Experimental Therapeutics. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 341, n. 3, p. 820-828, 2012.10.1124/jpet.112.1929970022-3565http://repositorio.unifesp.br/handle/11600/34918WOS:000304445000028engJournal of Pharmacology and Experimental Therapeuticsinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2016-01-24T12:27:17Zoai:repositorio.unifesp.br/:11600/34918Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652016-01-24T12:27:17Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
title Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
spellingShingle Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
Duarte, Thiago [UNIFESP]
title_short Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
title_full Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
title_fullStr Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
title_full_unstemmed Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
title_sort Contribution of the Extracellular cAMP-Adenosine Pathway to Dual Coupling of beta(2)-Adrenoceptors to G(s) and G(i) Proteins in Mouse Skeletal Muscle
author Duarte, Thiago [UNIFESP]
author_facet Duarte, Thiago [UNIFESP]
Menezes-Rodrigues, Francisco Sandro [UNIFESP]
Godinho, Rosely Oliveira [UNIFESP]
author_role author
author2 Menezes-Rodrigues, Francisco Sandro [UNIFESP]
Godinho, Rosely Oliveira [UNIFESP]
author2_role author
author
dc.contributor.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Duarte, Thiago [UNIFESP]
Menezes-Rodrigues, Francisco Sandro [UNIFESP]
Godinho, Rosely Oliveira [UNIFESP]
description beta(2)-Adrenoceptor (beta(2)-AR) agonists increase skeletal muscle contractile force via activation of G(s) protein/adenylyl cyclases (AC) and increased generation of cAMP. Herein, we evaluated the possible dual coupling of beta(2)-AR to G(s) and G(i) proteins and the influence of the beta(2)-AR/G(s)-G(i) /cAMP signaling cascade on skeletal muscle contraction. Assuming that the increment of intracellular cAMP is followed by cAMP efflux and extracellular generation of adenosine, the contribution of the extracellular cAMP-adenosine pathway on the beta(2)-AR inotropic response was also addressed. the effects of clenbuterol/fenoterol (beta(2)-AR agonists), forskolin (AC activator), cAMP/8-bromo-cAMP, and adenosine were evaluated on isometric contractility of mouse diaphragm muscle induced by supramaximal direct electrical stimulation (0.1 Hz, 2 ms duration). Clenbuterol/fenoterol (10-1000 mu M), 1 mu M forskolin, and 20 mu M rolipram induced transient positive inotropic effects that peaked 30 min after stimulation onset, declining to 10 to 20% of peak levels in 30 min. the late descending phase of the beta(2)-AR agonist inotropic effect was mimicked by either cAMP or adenosine and abolished by preincubation of diaphragm with pertussis toxin (PTX) (G(i) signaling inhibitor) or the organic anion transporter inhibitor probenecid, indicating a delayed coupling of beta(2)-AR to G(i) protein which depends on cAMP efflux. Remarkably, the PTX-sensitive beta(2)-AR inotropic effect was inhibited by the A(1) adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine and ecto-5'-phosphodiesterase inhibitor alpha,beta-methyleneadenosine 5'-diphosphate sodium salt, indicating that beta(2)-AR coupling to G(i) is indirect and dependent on A(1) receptor activation. the involvement of the extracellular cAMP-adenosine pathway in beta(2)-AR signaling would provide a negative feedback loop that may limit stimulatory G protein-coupled receptor positive inotropism and potential deleterious effects of excessive contractile response.
publishDate 2012
dc.date.none.fl_str_mv 2012-06-01
2016-01-24T14:27:17Z
2016-01-24T14:27:17Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1124/jpet.112.192997
Journal of Pharmacology and Experimental Therapeutics. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 341, n. 3, p. 820-828, 2012.
10.1124/jpet.112.192997
0022-3565
http://repositorio.unifesp.br/handle/11600/34918
WOS:000304445000028
url http://dx.doi.org/10.1124/jpet.112.192997
http://repositorio.unifesp.br/handle/11600/34918
identifier_str_mv Journal of Pharmacology and Experimental Therapeutics. Bethesda: Amer Soc Pharmacology Experimental Therapeutics, v. 341, n. 3, p. 820-828, 2012.
10.1124/jpet.112.192997
0022-3565
WOS:000304445000028
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Pharmacology and Experimental Therapeutics
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 820-828
dc.publisher.none.fl_str_mv Amer Soc Pharmacology Experimental Therapeutics
publisher.none.fl_str_mv Amer Soc Pharmacology Experimental Therapeutics
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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