Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells

Autores
Ronda, Ana Carolina; Boland, Ricardo Leopoldo
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2.GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblastsand myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptorwas present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypanblue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, wedemonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2.However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of themolecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptoticconditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps ofC2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments inundifferentiated and differentiated C2C12 cells and mediates E2 actions.
Fil: Ronda, Ana Carolina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; Argentina
Materia
Gpr30
Estradiol
Myoblasts
Myotubes
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:11336/6345
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 CellsRonda, Ana CarolinaBoland, Ricardo LeopoldoGpr30EstradiolMyoblastsMyotubeshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2.GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblastsand myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptorwas present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypanblue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, wedemonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2.However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of themolecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptoticconditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps ofC2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments inundifferentiated and differentiated C2C12 cells and mediates E2 actions.Fil: Ronda, Ana Carolina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; ArgentinaFil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; ArgentinaWiley2015-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/6345Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells; Wiley; Journal of Cellular Biochemistry; 117; 3; 9-2015; 793–8050730-2312enginfo:eu-repo/semantics/altIdentifier/doi/info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.25369info:eu-repo/semantics/altIdentifier/pmid/26359786info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/wol1/doi/10.1002/jcb.25369/abstractinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:10:05Zoai:ri.conicet.gov.ar:11336/6345instacron: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 10:10:05.392CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
title Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
spellingShingle Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
Ronda, Ana Carolina
Gpr30
Estradiol
Myoblasts
Myotubes
title_short Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
title_full Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
title_fullStr Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
title_full_unstemmed Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
title_sort Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells
dc.creator.none.fl_str_mv Ronda, Ana Carolina
Boland, Ricardo Leopoldo
author Ronda, Ana Carolina
author_facet Ronda, Ana Carolina
Boland, Ricardo Leopoldo
author_role author
author2 Boland, Ricardo Leopoldo
author2_role author
dc.subject.none.fl_str_mv Gpr30
Estradiol
Myoblasts
Myotubes
topic Gpr30
Estradiol
Myoblasts
Myotubes
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2.GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblastsand myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptorwas present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypanblue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, wedemonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2.However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of themolecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptoticconditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps ofC2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments inundifferentiated and differentiated C2C12 cells and mediates E2 actions.
Fil: Ronda, Ana Carolina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico Bahia Blanca. Instituto de Ciencias Biologicas y Biomedicas del Sur; Argentina. Universidad Nacional del Sur; Argentina
description G-protein-coupled receptor 30 (GPR30) is an estrogen receptor that initiates several rapid, non-genomic signaling events triggered by E2.GPR30 has recently been identified in C2C12 cells; however, little is known about the intracelular distribution and its role in C2C12 myoblastsand myotubes. By western blotting and immunohistochemistry, we evidenced expression of GPR30. While in C2C12 myoblasts, the receptorwas present in nucleus, mitochondria, and endoplasmic reticulum, in C2C12 myotubes, it was additionally found in cytoplasm. Using trypanblue uptake assay to determine cellular death and fluorescent microscopy to evaluate picnotic nuclei and mitochondrial distribution, wedemonstated that treatment of C2C12 myoblasts with G1 (GPR30 agonist) did not protect the cells against apoptosis induced by H2O2 as E2.However, when G15 (GPR30 antagonist) was used, E2 could not prevent the damage caused by the oxidative stress. Further, some of themolecular mechanisms involved were investigated by wertern blot assays. Thus, E2 was able to induce AKT phosphorylation in apoptoticconditions and ERK phosphorylation in proliferating C2C12 cells but not when the cultures were incubated with G15. Additionally, using G15antagonist we have found that GPR30 participates in the myogenin expression and creatine kinase activity stimulated by E2 in the first steps ofC2C12 differentiation. Althogether these findings provide evidences showing that GPR30 is expressed in diverse intracellular compartments inundifferentiated and differentiated C2C12 cells and mediates E2 actions.
publishDate 2015
dc.date.none.fl_str_mv 2015-09
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/11336/6345
Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells; Wiley; Journal of Cellular Biochemistry; 117; 3; 9-2015; 793–805
0730-2312
url http://hdl.handle.net/11336/6345
identifier_str_mv Ronda, Ana Carolina; Boland, Ricardo Leopoldo; Intracellular Distribution and Involvement of GPR30 in the Actions of E2 on C2C12 Cells; Wiley; Journal of Cellular Biochemistry; 117; 3; 9-2015; 793–805
0730-2312
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/
info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.25369
info:eu-repo/semantics/altIdentifier/pmid/26359786
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/wol1/doi/10.1002/jcb.25369/abstract
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
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
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score 13.13397

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