ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein

Autores
Assef, Yanina Andrea; Ozu, Marcelo; Marino, Gabriela Inés; Galizia, Luciano; Kotsias, Basilio Aristides
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Shroom is a family of related proteins linked to the actin cytoskeleton. xShroom1 is constitutively expressed in X. oocytes and is required for the expression of amiloride sensitive sodium channels (ENaC). Oocytes were injected with a, b, and g mENaC and xShroom1 sense or antisense oligonucleotides. We used voltage clamp techniques to study the amiloride-sensitive Na(+) currents (INa((amil))). We observed a marked reduction in INa((amil)) in oocytes co-injected with xShroom1 antisense. Oocytes expressing a DEG mutant â-mENaC subunit (â-S518K) with an open probability of 1 had enhanced INa((amil)) although these currents were also reduced when co-injected with xShroom1 antisense. Addition of low concentration (20 ng/ml) of trypsin which activates the membrane-resident ENaC channels led to a slow increase in INa((amil)) in oocytes with xShroom1 sense but had no effect on the currents in oocytes coinjected with ENaC and xShroom1 antisense. The same results were obtained with higher concentrations of trypsin (2 ug/ml) exposed during 2.5 min. In addition, fluorescence positive staining of plasma membrane in the oocytes expressing a, b and g mENaC and xShroom1 sense were observed but not in oocytes coinjected with ENaC and xShroom1 antisense oligonucleotides. On this basis, we suggest that xShroom1-dependent ENaC inhibition may be through the number of channels inserted in the membrane.
Fil: Assef, Yanina Andrea. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ozu, Marcelo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marino, Gabriela Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina
Fil: Galizia, Luciano. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Kotsias, Basilio Aristides. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
ENaC
Sodium currents
xShroom1
Oocytes
Trypsin
Immunofluoresence
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:11336/236644
Acceder
id CONICETDig_e5819bc02f190a840508a438416aa3b9
oai_identifier_str oai:ri.conicet.gov.ar:11336/236644
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 proteinAssef, Yanina AndreaOzu, MarceloMarino, Gabriela InésGalizia, LucianoKotsias, Basilio AristidesENaCSodium currentsxShroom1OocytesTrypsinImmunofluoresencehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Shroom is a family of related proteins linked to the actin cytoskeleton. xShroom1 is constitutively expressed in X. oocytes and is required for the expression of amiloride sensitive sodium channels (ENaC). Oocytes were injected with a, b, and g mENaC and xShroom1 sense or antisense oligonucleotides. We used voltage clamp techniques to study the amiloride-sensitive Na(+) currents (INa((amil))). We observed a marked reduction in INa((amil)) in oocytes co-injected with xShroom1 antisense. Oocytes expressing a DEG mutant â-mENaC subunit (â-S518K) with an open probability of 1 had enhanced INa((amil)) although these currents were also reduced when co-injected with xShroom1 antisense. Addition of low concentration (20 ng/ml) of trypsin which activates the membrane-resident ENaC channels led to a slow increase in INa((amil)) in oocytes with xShroom1 sense but had no effect on the currents in oocytes coinjected with ENaC and xShroom1 antisense. The same results were obtained with higher concentrations of trypsin (2 ug/ml) exposed during 2.5 min. In addition, fluorescence positive staining of plasma membrane in the oocytes expressing a, b and g mENaC and xShroom1 sense were observed but not in oocytes coinjected with ENaC and xShroom1 antisense oligonucleotides. On this basis, we suggest that xShroom1-dependent ENaC inhibition may be through the number of channels inserted in the membrane.Fil: Assef, Yanina Andrea. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ozu, Marcelo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marino, Gabriela Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Galizia, Luciano. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Kotsias, Basilio Aristides. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaKarger2011-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/236644Assef, Yanina Andrea; Ozu, Marcelo; Marino, Gabriela Inés; Galizia, Luciano; Kotsias, Basilio Aristides; ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein; Karger; Cellular Physiology and Biochemistry; 28; 2; 8-2011; 259-2661015-89871421-9778CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1159/000331738info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:10:39Zoai:ri.conicet.gov.ar:11336/236644instacron: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 10:10:40.241CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
title ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
spellingShingle ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
Assef, Yanina Andrea
ENaC
Sodium currents
xShroom1
Oocytes
Trypsin
Immunofluoresence
title_short ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
title_full ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
title_fullStr ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
title_full_unstemmed ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
title_sort ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein
dc.creator.none.fl_str_mv Assef, Yanina Andrea
Ozu, Marcelo
Marino, Gabriela Inés
Galizia, Luciano
Kotsias, Basilio Aristides
author Assef, Yanina Andrea
author_facet Assef, Yanina Andrea
Ozu, Marcelo
Marino, Gabriela Inés
Galizia, Luciano
Kotsias, Basilio Aristides
author_role author
author2 Ozu, Marcelo
Marino, Gabriela Inés
Galizia, Luciano
Kotsias, Basilio Aristides
author2_role author
author
author
author
dc.subject.none.fl_str_mv ENaC
Sodium currents
xShroom1
Oocytes
Trypsin
Immunofluoresence
topic ENaC
Sodium currents
xShroom1
Oocytes
Trypsin
Immunofluoresence
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Shroom is a family of related proteins linked to the actin cytoskeleton. xShroom1 is constitutively expressed in X. oocytes and is required for the expression of amiloride sensitive sodium channels (ENaC). Oocytes were injected with a, b, and g mENaC and xShroom1 sense or antisense oligonucleotides. We used voltage clamp techniques to study the amiloride-sensitive Na(+) currents (INa((amil))). We observed a marked reduction in INa((amil)) in oocytes co-injected with xShroom1 antisense. Oocytes expressing a DEG mutant â-mENaC subunit (â-S518K) with an open probability of 1 had enhanced INa((amil)) although these currents were also reduced when co-injected with xShroom1 antisense. Addition of low concentration (20 ng/ml) of trypsin which activates the membrane-resident ENaC channels led to a slow increase in INa((amil)) in oocytes with xShroom1 sense but had no effect on the currents in oocytes coinjected with ENaC and xShroom1 antisense. The same results were obtained with higher concentrations of trypsin (2 ug/ml) exposed during 2.5 min. In addition, fluorescence positive staining of plasma membrane in the oocytes expressing a, b and g mENaC and xShroom1 sense were observed but not in oocytes coinjected with ENaC and xShroom1 antisense oligonucleotides. On this basis, we suggest that xShroom1-dependent ENaC inhibition may be through the number of channels inserted in the membrane.
Fil: Assef, Yanina Andrea. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ozu, Marcelo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marino, Gabriela Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina
Fil: Galizia, Luciano. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Kotsias, Basilio Aristides. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Shroom is a family of related proteins linked to the actin cytoskeleton. xShroom1 is constitutively expressed in X. oocytes and is required for the expression of amiloride sensitive sodium channels (ENaC). Oocytes were injected with a, b, and g mENaC and xShroom1 sense or antisense oligonucleotides. We used voltage clamp techniques to study the amiloride-sensitive Na(+) currents (INa((amil))). We observed a marked reduction in INa((amil)) in oocytes co-injected with xShroom1 antisense. Oocytes expressing a DEG mutant â-mENaC subunit (â-S518K) with an open probability of 1 had enhanced INa((amil)) although these currents were also reduced when co-injected with xShroom1 antisense. Addition of low concentration (20 ng/ml) of trypsin which activates the membrane-resident ENaC channels led to a slow increase in INa((amil)) in oocytes with xShroom1 sense but had no effect on the currents in oocytes coinjected with ENaC and xShroom1 antisense. The same results were obtained with higher concentrations of trypsin (2 ug/ml) exposed during 2.5 min. In addition, fluorescence positive staining of plasma membrane in the oocytes expressing a, b and g mENaC and xShroom1 sense were observed but not in oocytes coinjected with ENaC and xShroom1 antisense oligonucleotides. On this basis, we suggest that xShroom1-dependent ENaC inhibition may be through the number of channels inserted in the membrane.
publishDate 2011
dc.date.none.fl_str_mv 2011-08
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/11336/236644
Assef, Yanina Andrea; Ozu, Marcelo; Marino, Gabriela Inés; Galizia, Luciano; Kotsias, Basilio Aristides; ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein; Karger; Cellular Physiology and Biochemistry; 28; 2; 8-2011; 259-266
1015-8987
1421-9778
CONICET Digital
CONICET
url http://hdl.handle.net/11336/236644
identifier_str_mv Assef, Yanina Andrea; Ozu, Marcelo; Marino, Gabriela Inés; Galizia, Luciano; Kotsias, Basilio Aristides; ENaC channels in oocytes from Xenopus laevis and their regulation by xShroom1 protein; Karger; Cellular Physiology and Biochemistry; 28; 2; 8-2011; 259-266
1015-8987
1421-9778
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1159/000331738
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
application/pdf
dc.publisher.none.fl_str_mv Karger
publisher.none.fl_str_mv Karger
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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