A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer

Autores
Pafundo, Diego Esteban; Alvarez, Cora Lilia; Krumschnabel, Gerhard; Schwarzbaum, Pablo Julio
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Human erythrocytes have been regarded as perfect osmometers, which swell or shrink as dictated by their osmotic environment. In contrast, in most other cells, swelling elicits a regulatory volume decrease (RVD) modulated by the activation of purinic and pyrimidinic receptors (P receptors). For human erythrocytes this modulation has not been tested, and we thus investigated whether P receptor activation can induce RVD in these cells. Further, because ectonucleotidases may scavenge ATP or ADP or act as a source for extracellular adenosine and therefore modulate P receptor activation and RVD, we also determined their activity in intact erythrocytes. We found relatively low ectoATPase but significant ectoADPase and ectoAMPase activities. When erythrocytes were exposed to hypotonic medium alone, they swelled as expected for an osmometric response and showed no RVD. Activation of P2 receptors by exogenous ATP or ADP did not trigger RVD, whereas P1 agonists adenosine and adenosine-5′-N-ethylcarboxamide induced significant RVD. The effect of adenosine-5′-N-ethylcarboxamide was dose-dependent (maximal RVD of 27%; apparent K½ of 1.6 ± 1.7 μM). The RVD induced by adenosine was blocked 80% with the non-selective P1 antagonist 8-(p-sulfophenyl theophylline) or the P1-A2B inhibitor MRS1754, but not by inhibitors of P1 subtypes A1, A2A, and A3. In addition, forskolin (an inducer of intracellular cAMP formation) could mimic the effect of adenosine, supporting the idea of P1-A2B receptor activation. In conclusion, we report a novel P1-A2B receptor-mediated RVD activation in mature human erythrocytes and thus indicate that these long held perfect osmometers are not so perfect after all.
Fil: Pafundo, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Krumschnabel, Gerhard. Universidad de Innsbruck; Austria
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Materia
RVD
Volume regulation
Water transport
Nucleotide metabolism
Cell/Blood
Cellular Regulation
Erythrocyte
Purinergic Agonists
Purinergic Receptor
P1 Receptor
Extracellular Adenosine
Fluorescence
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/18250
Acceder
id CONICETDig_5cd0b4e7147fe6c4467fc0887a03a500
oai_identifier_str oai:ri.conicet.gov.ar:11336/18250
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longerPafundo, Diego EstebanAlvarez, Cora LiliaKrumschnabel, GerhardSchwarzbaum, Pablo JulioRVDVolume regulationWater transportNucleotide metabolismCell/BloodCellular RegulationErythrocytePurinergic AgonistsPurinergic ReceptorP1 ReceptorExtracellular AdenosineFluorescencehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Human erythrocytes have been regarded as perfect osmometers, which swell or shrink as dictated by their osmotic environment. In contrast, in most other cells, swelling elicits a regulatory volume decrease (RVD) modulated by the activation of purinic and pyrimidinic receptors (P receptors). For human erythrocytes this modulation has not been tested, and we thus investigated whether P receptor activation can induce RVD in these cells. Further, because ectonucleotidases may scavenge ATP or ADP or act as a source for extracellular adenosine and therefore modulate P receptor activation and RVD, we also determined their activity in intact erythrocytes. We found relatively low ectoATPase but significant ectoADPase and ectoAMPase activities. When erythrocytes were exposed to hypotonic medium alone, they swelled as expected for an osmometric response and showed no RVD. Activation of P2 receptors by exogenous ATP or ADP did not trigger RVD, whereas P1 agonists adenosine and adenosine-5′-N-ethylcarboxamide induced significant RVD. The effect of adenosine-5′-N-ethylcarboxamide was dose-dependent (maximal RVD of 27%; apparent K½ of 1.6 ± 1.7 μM). The RVD induced by adenosine was blocked 80% with the non-selective P1 antagonist 8-(p-sulfophenyl theophylline) or the P1-A2B inhibitor MRS1754, but not by inhibitors of P1 subtypes A1, A2A, and A3. In addition, forskolin (an inducer of intracellular cAMP formation) could mimic the effect of adenosine, supporting the idea of P1-A2B receptor activation. In conclusion, we report a novel P1-A2B receptor-mediated RVD activation in mature human erythrocytes and thus indicate that these long held perfect osmometers are not so perfect after all.Fil: Pafundo, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Krumschnabel, Gerhard. Universidad de Innsbruck; AustriaFil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaAmerican Society for Biochemistry and Molecular Biology2010-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/18250Pafundo, Diego Esteban; Alvarez, Cora Lilia; Krumschnabel, Gerhard; Schwarzbaum, Pablo Julio; A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 285; 9; 2-2010; 6134-61440021-92581083-351XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M109.078246info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/285/9/6134.longinfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2825408/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:17:12Zoai:ri.conicet.gov.ar:11336/18250instacron: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:34982025-09-10 13:17:12.661CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
title A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
spellingShingle A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
Pafundo, Diego Esteban
RVD
Volume regulation
Water transport
Nucleotide metabolism
Cell/Blood
Cellular Regulation
Erythrocyte
Purinergic Agonists
Purinergic Receptor
P1 Receptor
Extracellular Adenosine
Fluorescence
title_short A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
title_full A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
title_fullStr A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
title_full_unstemmed A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
title_sort A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer
dc.creator.none.fl_str_mv Pafundo, Diego Esteban
Alvarez, Cora Lilia
Krumschnabel, Gerhard
Schwarzbaum, Pablo Julio
author Pafundo, Diego Esteban
author_facet Pafundo, Diego Esteban
Alvarez, Cora Lilia
Krumschnabel, Gerhard
Schwarzbaum, Pablo Julio
author_role author
author2 Alvarez, Cora Lilia
Krumschnabel, Gerhard
Schwarzbaum, Pablo Julio
author2_role author
author
author
dc.subject.none.fl_str_mv RVD
Volume regulation
Water transport
Nucleotide metabolism
Cell/Blood
Cellular Regulation
Erythrocyte
Purinergic Agonists
Purinergic Receptor
P1 Receptor
Extracellular Adenosine
Fluorescence
topic RVD
Volume regulation
Water transport
Nucleotide metabolism
Cell/Blood
Cellular Regulation
Erythrocyte
Purinergic Agonists
Purinergic Receptor
P1 Receptor
Extracellular Adenosine
Fluorescence
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Human erythrocytes have been regarded as perfect osmometers, which swell or shrink as dictated by their osmotic environment. In contrast, in most other cells, swelling elicits a regulatory volume decrease (RVD) modulated by the activation of purinic and pyrimidinic receptors (P receptors). For human erythrocytes this modulation has not been tested, and we thus investigated whether P receptor activation can induce RVD in these cells. Further, because ectonucleotidases may scavenge ATP or ADP or act as a source for extracellular adenosine and therefore modulate P receptor activation and RVD, we also determined their activity in intact erythrocytes. We found relatively low ectoATPase but significant ectoADPase and ectoAMPase activities. When erythrocytes were exposed to hypotonic medium alone, they swelled as expected for an osmometric response and showed no RVD. Activation of P2 receptors by exogenous ATP or ADP did not trigger RVD, whereas P1 agonists adenosine and adenosine-5′-N-ethylcarboxamide induced significant RVD. The effect of adenosine-5′-N-ethylcarboxamide was dose-dependent (maximal RVD of 27%; apparent K½ of 1.6 ± 1.7 μM). The RVD induced by adenosine was blocked 80% with the non-selective P1 antagonist 8-(p-sulfophenyl theophylline) or the P1-A2B inhibitor MRS1754, but not by inhibitors of P1 subtypes A1, A2A, and A3. In addition, forskolin (an inducer of intracellular cAMP formation) could mimic the effect of adenosine, supporting the idea of P1-A2B receptor activation. In conclusion, we report a novel P1-A2B receptor-mediated RVD activation in mature human erythrocytes and thus indicate that these long held perfect osmometers are not so perfect after all.
Fil: Pafundo, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Krumschnabel, Gerhard. Universidad de Innsbruck; Austria
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
description Human erythrocytes have been regarded as perfect osmometers, which swell or shrink as dictated by their osmotic environment. In contrast, in most other cells, swelling elicits a regulatory volume decrease (RVD) modulated by the activation of purinic and pyrimidinic receptors (P receptors). For human erythrocytes this modulation has not been tested, and we thus investigated whether P receptor activation can induce RVD in these cells. Further, because ectonucleotidases may scavenge ATP or ADP or act as a source for extracellular adenosine and therefore modulate P receptor activation and RVD, we also determined their activity in intact erythrocytes. We found relatively low ectoATPase but significant ectoADPase and ectoAMPase activities. When erythrocytes were exposed to hypotonic medium alone, they swelled as expected for an osmometric response and showed no RVD. Activation of P2 receptors by exogenous ATP or ADP did not trigger RVD, whereas P1 agonists adenosine and adenosine-5′-N-ethylcarboxamide induced significant RVD. The effect of adenosine-5′-N-ethylcarboxamide was dose-dependent (maximal RVD of 27%; apparent K½ of 1.6 ± 1.7 μM). The RVD induced by adenosine was blocked 80% with the non-selective P1 antagonist 8-(p-sulfophenyl theophylline) or the P1-A2B inhibitor MRS1754, but not by inhibitors of P1 subtypes A1, A2A, and A3. In addition, forskolin (an inducer of intracellular cAMP formation) could mimic the effect of adenosine, supporting the idea of P1-A2B receptor activation. In conclusion, we report a novel P1-A2B receptor-mediated RVD activation in mature human erythrocytes and thus indicate that these long held perfect osmometers are not so perfect after all.
publishDate 2010
dc.date.none.fl_str_mv 2010-02
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/11336/18250
Pafundo, Diego Esteban; Alvarez, Cora Lilia; Krumschnabel, Gerhard; Schwarzbaum, Pablo Julio; A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 285; 9; 2-2010; 6134-6144
0021-9258
1083-351X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/18250
identifier_str_mv Pafundo, Diego Esteban; Alvarez, Cora Lilia; Krumschnabel, Gerhard; Schwarzbaum, Pablo Julio; A volume regulatory response can be triggered by nucleosides in human erythrocytes, a perfect osmometer no longer; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry; 285; 9; 2-2010; 6134-6144
0021-9258
1083-351X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M109.078246
info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/285/9/6134.long
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2825408/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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 12.993085

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