Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells

Autores
Orlowski, Alejandro; Vargas, Lorena Alejandra; Aiello, Ernesto Alejandro; Alvarez, Bernardo
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The NBCn1 Na+/HCO3− cotransporter catalyzes the electroneutral movement of 1 Na+:1 HCO3− into kidney cells. We characterized the intracellular pH (pHi) regulation in human embryonic kidney cells (HEK) subjected to NH4Cl prepulse acid loading, and we examined the NBCn1 expression and function in HEK cells subjected to 24-h elevated Pco2 (10–15%). After acid loading, in the presence of HCO3−, ∼50% of the pHi recovery phase was blocked by the Na+/H+ exchanger inhibitors EIPA (10–50 μM) and amiloride (1 mM) and was fully cancelled by 30 μM EIPA under nominally HCO3−-free conditions. In addition, in the presence of HCO3−, pHi recovery after acid loading was completely blocked when Na+ was omitted in the buffer. pHi recovery after acidification in HEK cells was repeated in the presence of the NBC inhibitor S0859, and the pHi recovery was inhibited by S0859 in a dose-dependent manner (Ki = 30 μM, full inhibition at 60 μM), which confirmed NBC Na+/HCO3− cotransporter activation. NBCn1 expression increased threefold after 24-h exposure of cultured HEK cells to 10% CO2 and sevenfold after exposure to 15% CO2, examined by immunoblots. Finally, exposure of HEK cells to high CO2 significantly increased the HCO3−-dependent recovery of pHi after acid loading. We conclude that HEK cells expressed the NBCn1 Na+/HCO3− cotransporter as the only HCO3−-dependent mechanism responsible for cellular alkaline loading. NBCn1, which expresses in different kidney cell types, was upregulated by 24-h high-Pco2 exposure of HEK cells, and this upregulation was accompanied by increased NBCn1-mediated HCO3− transport.
Fil: Orlowski, Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Vargas, Lorena Alejandra. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Aiello, Ernesto Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Alvarez, Bernardo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Materia
Nbcn1 Na+/Hco3− Cotransporter
Human Embryonic Kidney Cells
High Pco2
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:11336/11957
Acceder
id CONICETDig_95a9402679a6d6aef0ded48011186630
oai_identifier_str oai:ri.conicet.gov.ar:11336/11957
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cellsOrlowski, AlejandroVargas, Lorena AlejandraAiello, Ernesto AlejandroAlvarez, BernardoNbcn1 Na+/Hco3− CotransporterHuman Embryonic Kidney CellsHigh Pco2https://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3The NBCn1 Na+/HCO3− cotransporter catalyzes the electroneutral movement of 1 Na+:1 HCO3− into kidney cells. We characterized the intracellular pH (pHi) regulation in human embryonic kidney cells (HEK) subjected to NH4Cl prepulse acid loading, and we examined the NBCn1 expression and function in HEK cells subjected to 24-h elevated Pco2 (10–15%). After acid loading, in the presence of HCO3−, ∼50% of the pHi recovery phase was blocked by the Na+/H+ exchanger inhibitors EIPA (10–50 μM) and amiloride (1 mM) and was fully cancelled by 30 μM EIPA under nominally HCO3−-free conditions. In addition, in the presence of HCO3−, pHi recovery after acid loading was completely blocked when Na+ was omitted in the buffer. pHi recovery after acidification in HEK cells was repeated in the presence of the NBC inhibitor S0859, and the pHi recovery was inhibited by S0859 in a dose-dependent manner (Ki = 30 μM, full inhibition at 60 μM), which confirmed NBC Na+/HCO3− cotransporter activation. NBCn1 expression increased threefold after 24-h exposure of cultured HEK cells to 10% CO2 and sevenfold after exposure to 15% CO2, examined by immunoblots. Finally, exposure of HEK cells to high CO2 significantly increased the HCO3−-dependent recovery of pHi after acid loading. We conclude that HEK cells expressed the NBCn1 Na+/HCO3− cotransporter as the only HCO3−-dependent mechanism responsible for cellular alkaline loading. NBCn1, which expresses in different kidney cell types, was upregulated by 24-h high-Pco2 exposure of HEK cells, and this upregulation was accompanied by increased NBCn1-mediated HCO3− transport.Fil: Orlowski, Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Vargas, Lorena Alejandra. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Aiello, Ernesto Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaFil: Alvarez, Bernardo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; ArgentinaAmerican Physiological Society2013-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/mswordapplication/pdfhttp://hdl.handle.net/11336/11957Orlowski, Alejandro; Vargas, Lorena Alejandra; Aiello, Ernesto Alejandro; Alvarez, Bernardo; Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 305; 12; 12-2013; 1765-17741931-857X1522-1466enginfo:eu-repo/semantics/altIdentifier/doi/10.1152/ajprenal.00096.2013info:eu-repo/semantics/altIdentifier/url/http://ajprenal.physiology.org/content/305/12/F1765info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-17T11:20:07Zoai:ri.conicet.gov.ar:11336/11957instacron: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:34982025-09-17 11:20:07.649CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
title Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
spellingShingle Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
Orlowski, Alejandro
Nbcn1 Na+/Hco3− Cotransporter
Human Embryonic Kidney Cells
High Pco2
title_short Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
title_full Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
title_fullStr Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
title_full_unstemmed Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
title_sort Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells
dc.creator.none.fl_str_mv Orlowski, Alejandro
Vargas, Lorena Alejandra
Aiello, Ernesto Alejandro
Alvarez, Bernardo
author Orlowski, Alejandro
author_facet Orlowski, Alejandro
Vargas, Lorena Alejandra
Aiello, Ernesto Alejandro
Alvarez, Bernardo
author_role author
author2 Vargas, Lorena Alejandra
Aiello, Ernesto Alejandro
Alvarez, Bernardo
author2_role author
author
author
dc.subject.none.fl_str_mv Nbcn1 Na+/Hco3− Cotransporter
Human Embryonic Kidney Cells
High Pco2
topic Nbcn1 Na+/Hco3− Cotransporter
Human Embryonic Kidney Cells
High Pco2
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv The NBCn1 Na+/HCO3− cotransporter catalyzes the electroneutral movement of 1 Na+:1 HCO3− into kidney cells. We characterized the intracellular pH (pHi) regulation in human embryonic kidney cells (HEK) subjected to NH4Cl prepulse acid loading, and we examined the NBCn1 expression and function in HEK cells subjected to 24-h elevated Pco2 (10–15%). After acid loading, in the presence of HCO3−, ∼50% of the pHi recovery phase was blocked by the Na+/H+ exchanger inhibitors EIPA (10–50 μM) and amiloride (1 mM) and was fully cancelled by 30 μM EIPA under nominally HCO3−-free conditions. In addition, in the presence of HCO3−, pHi recovery after acid loading was completely blocked when Na+ was omitted in the buffer. pHi recovery after acidification in HEK cells was repeated in the presence of the NBC inhibitor S0859, and the pHi recovery was inhibited by S0859 in a dose-dependent manner (Ki = 30 μM, full inhibition at 60 μM), which confirmed NBC Na+/HCO3− cotransporter activation. NBCn1 expression increased threefold after 24-h exposure of cultured HEK cells to 10% CO2 and sevenfold after exposure to 15% CO2, examined by immunoblots. Finally, exposure of HEK cells to high CO2 significantly increased the HCO3−-dependent recovery of pHi after acid loading. We conclude that HEK cells expressed the NBCn1 Na+/HCO3− cotransporter as the only HCO3−-dependent mechanism responsible for cellular alkaline loading. NBCn1, which expresses in different kidney cell types, was upregulated by 24-h high-Pco2 exposure of HEK cells, and this upregulation was accompanied by increased NBCn1-mediated HCO3− transport.
Fil: Orlowski, Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Vargas, Lorena Alejandra. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Aiello, Ernesto Alejandro. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
Fil: Alvarez, Bernardo. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnológico la Plata. Centro de Investigaciones Cardiovasculares "dr. Horacio Eugenio Cingolani"; Argentina. Universidad Nacional de la Plata. Facultad de Ciencias Médicas; Argentina
description The NBCn1 Na+/HCO3− cotransporter catalyzes the electroneutral movement of 1 Na+:1 HCO3− into kidney cells. We characterized the intracellular pH (pHi) regulation in human embryonic kidney cells (HEK) subjected to NH4Cl prepulse acid loading, and we examined the NBCn1 expression and function in HEK cells subjected to 24-h elevated Pco2 (10–15%). After acid loading, in the presence of HCO3−, ∼50% of the pHi recovery phase was blocked by the Na+/H+ exchanger inhibitors EIPA (10–50 μM) and amiloride (1 mM) and was fully cancelled by 30 μM EIPA under nominally HCO3−-free conditions. In addition, in the presence of HCO3−, pHi recovery after acid loading was completely blocked when Na+ was omitted in the buffer. pHi recovery after acidification in HEK cells was repeated in the presence of the NBC inhibitor S0859, and the pHi recovery was inhibited by S0859 in a dose-dependent manner (Ki = 30 μM, full inhibition at 60 μM), which confirmed NBC Na+/HCO3− cotransporter activation. NBCn1 expression increased threefold after 24-h exposure of cultured HEK cells to 10% CO2 and sevenfold after exposure to 15% CO2, examined by immunoblots. Finally, exposure of HEK cells to high CO2 significantly increased the HCO3−-dependent recovery of pHi after acid loading. We conclude that HEK cells expressed the NBCn1 Na+/HCO3− cotransporter as the only HCO3−-dependent mechanism responsible for cellular alkaline loading. NBCn1, which expresses in different kidney cell types, was upregulated by 24-h high-Pco2 exposure of HEK cells, and this upregulation was accompanied by increased NBCn1-mediated HCO3− transport.
publishDate 2013
dc.date.none.fl_str_mv 2013-12
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/11336/11957
Orlowski, Alejandro; Vargas, Lorena Alejandra; Aiello, Ernesto Alejandro; Alvarez, Bernardo; Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 305; 12; 12-2013; 1765-1774
1931-857X
1522-1466
url http://hdl.handle.net/11336/11957
identifier_str_mv Orlowski, Alejandro; Vargas, Lorena Alejandra; Aiello, Ernesto Alejandro; Alvarez, Bernardo; Elevated carbon dioxide upregulates NBCn1 Na+/HCO3− cotransporter in human embryonic kidney cells; American Physiological Society; American Journal Of Physiology-renal Physiology; 305; 12; 12-2013; 1765-1774
1931-857X
1522-1466
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1152/ajprenal.00096.2013
info:eu-repo/semantics/altIdentifier/url/http://ajprenal.physiology.org/content/305/12/F1765
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/msword
application/pdf
dc.publisher.none.fl_str_mv American Physiological Society
publisher.none.fl_str_mv American Physiological Society
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 13.001348

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