Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line

Autores
Milanesi, Lorena Magdalena; Russo, Ana Josefa; Boland, Ricardo Leopoldo
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The classical model of 17β-estradiol action has been traditionally described to be mediated by the estrogen receptor (ER) localized exclusively in the nucleus. However, there is increasing functional evidence for extra nuclear localization of ER. We present biochemical, immunological and molecular data supporting mitochondrial-microsomal localization of ERα in the C2C12 skeletal muscle cell line. We first established [3H]17βestradiol binding characteristics in whole cells in culture. Specific and saturable [3H]17βestradiol binding sites of high affinity were then detected in mitochondrial fractions (Kd = 0.43 nM; Bmax = 572 fmol/mg protein). Immunocytological studies revealed that estrogen receptors mainly localize at the mitochondrial and perinuclear level. These results were also confirmed using fluorescent 17βestradiol-BSA conjugates. The immunoreactivity did not translocate into the nucleus by 17β-estradiol treatment. Western and Ligand blot approaches corroborated the non-classical localization. Expression and subcellular distribution of ERα proteins were confirmed in C2C12 cellstransfected with ERα siRNA and by RT-PCR employing specific primers. The non-classical distribution of native pools of ERα in skeletal muscle cells suggests an alternative mode of ER localization/function.
Fil: Milanesi, Lorena Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
Fil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
Materia
C2c12 Muscle Cells
Erα
Estrogen Receptor (Er)
Skeletal Muscle
Subcellular Distribution
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:11336/76238
Acceder
id CONICETDig_c720fde4b4bfa745221c307405e7db29
oai_identifier_str oai:ri.conicet.gov.ar:11336/76238
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell lineMilanesi, Lorena MagdalenaRusso, Ana JosefaBoland, Ricardo LeopoldoC2c12 Muscle CellsErαEstrogen Receptor (Er)Skeletal MuscleSubcellular Distributionhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The classical model of 17β-estradiol action has been traditionally described to be mediated by the estrogen receptor (ER) localized exclusively in the nucleus. However, there is increasing functional evidence for extra nuclear localization of ER. We present biochemical, immunological and molecular data supporting mitochondrial-microsomal localization of ERα in the C2C12 skeletal muscle cell line. We first established [3H]17βestradiol binding characteristics in whole cells in culture. Specific and saturable [3H]17βestradiol binding sites of high affinity were then detected in mitochondrial fractions (Kd = 0.43 nM; Bmax = 572 fmol/mg protein). Immunocytological studies revealed that estrogen receptors mainly localize at the mitochondrial and perinuclear level. These results were also confirmed using fluorescent 17βestradiol-BSA conjugates. The immunoreactivity did not translocate into the nucleus by 17β-estradiol treatment. Western and Ligand blot approaches corroborated the non-classical localization. Expression and subcellular distribution of ERα proteins were confirmed in C2C12 cellstransfected with ERα siRNA and by RT-PCR employing specific primers. The non-classical distribution of native pools of ERα in skeletal muscle cells suggests an alternative mode of ER localization/function.Fil: Milanesi, Lorena Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaWiley-liss, Div John Wiley & Sons Inc2008-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/76238Milanesi, Lorena Magdalena; Russo, Ana Josefa; Boland, Ricardo Leopoldo; Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line; Wiley-liss, Div John Wiley & Sons Inc; Journal of Cellular Biochemistry; 104; 4; 7-2008; 1254-12730730-23121097-4644CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1002/jcb.21706info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.21706info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:41:01Zoai:ri.conicet.gov.ar:11336/76238instacron: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:34982025-10-15 14:41:02.277CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
title Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
spellingShingle Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
Milanesi, Lorena Magdalena
C2c12 Muscle Cells
Erα
Estrogen Receptor (Er)
Skeletal Muscle
Subcellular Distribution
title_short Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
title_full Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
title_fullStr Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
title_full_unstemmed Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
title_sort Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line
dc.creator.none.fl_str_mv Milanesi, Lorena Magdalena
Russo, Ana Josefa
Boland, Ricardo Leopoldo
author Milanesi, Lorena Magdalena
author_facet Milanesi, Lorena Magdalena
Russo, Ana Josefa
Boland, Ricardo Leopoldo
author_role author
author2 Russo, Ana Josefa
Boland, Ricardo Leopoldo
author2_role author
author
dc.subject.none.fl_str_mv C2c12 Muscle Cells
Erα
Estrogen Receptor (Er)
Skeletal Muscle
Subcellular Distribution
topic C2c12 Muscle Cells
Erα
Estrogen Receptor (Er)
Skeletal Muscle
Subcellular Distribution
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 classical model of 17β-estradiol action has been traditionally described to be mediated by the estrogen receptor (ER) localized exclusively in the nucleus. However, there is increasing functional evidence for extra nuclear localization of ER. We present biochemical, immunological and molecular data supporting mitochondrial-microsomal localization of ERα in the C2C12 skeletal muscle cell line. We first established [3H]17βestradiol binding characteristics in whole cells in culture. Specific and saturable [3H]17βestradiol binding sites of high affinity were then detected in mitochondrial fractions (Kd = 0.43 nM; Bmax = 572 fmol/mg protein). Immunocytological studies revealed that estrogen receptors mainly localize at the mitochondrial and perinuclear level. These results were also confirmed using fluorescent 17βestradiol-BSA conjugates. The immunoreactivity did not translocate into the nucleus by 17β-estradiol treatment. Western and Ligand blot approaches corroborated the non-classical localization. Expression and subcellular distribution of ERα proteins were confirmed in C2C12 cellstransfected with ERα siRNA and by RT-PCR employing specific primers. The non-classical distribution of native pools of ERα in skeletal muscle cells suggests an alternative mode of ER localization/function.
Fil: Milanesi, Lorena Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
Fil: Russo, Ana Josefa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
Fil: Boland, Ricardo Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina
description The classical model of 17β-estradiol action has been traditionally described to be mediated by the estrogen receptor (ER) localized exclusively in the nucleus. However, there is increasing functional evidence for extra nuclear localization of ER. We present biochemical, immunological and molecular data supporting mitochondrial-microsomal localization of ERα in the C2C12 skeletal muscle cell line. We first established [3H]17βestradiol binding characteristics in whole cells in culture. Specific and saturable [3H]17βestradiol binding sites of high affinity were then detected in mitochondrial fractions (Kd = 0.43 nM; Bmax = 572 fmol/mg protein). Immunocytological studies revealed that estrogen receptors mainly localize at the mitochondrial and perinuclear level. These results were also confirmed using fluorescent 17βestradiol-BSA conjugates. The immunoreactivity did not translocate into the nucleus by 17β-estradiol treatment. Western and Ligand blot approaches corroborated the non-classical localization. Expression and subcellular distribution of ERα proteins were confirmed in C2C12 cellstransfected with ERα siRNA and by RT-PCR employing specific primers. The non-classical distribution of native pools of ERα in skeletal muscle cells suggests an alternative mode of ER localization/function.
publishDate 2008
dc.date.none.fl_str_mv 2008-07
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/11336/76238
Milanesi, Lorena Magdalena; Russo, Ana Josefa; Boland, Ricardo Leopoldo; Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line; Wiley-liss, Div John Wiley & Sons Inc; Journal of Cellular Biochemistry; 104; 4; 7-2008; 1254-1273
0730-2312
1097-4644
CONICET Digital
CONICET
url http://hdl.handle.net/11336/76238
identifier_str_mv Milanesi, Lorena Magdalena; Russo, Ana Josefa; Boland, Ricardo Leopoldo; Expression and localization of estrogen receptor α in the C2C12 murine skeletal muscle cell line; Wiley-liss, Div John Wiley & Sons Inc; Journal of Cellular Biochemistry; 104; 4; 7-2008; 1254-1273
0730-2312
1097-4644
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://onlinelibrary.wiley.com/doi/10.1002/jcb.21706
info:eu-repo/semantics/altIdentifier/doi/10.1002/jcb.21706
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
dc.publisher.none.fl_str_mv Wiley-liss, Div John Wiley & Sons Inc
publisher.none.fl_str_mv Wiley-liss, Div John Wiley & Sons Inc
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 13.22299

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