Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation

Autores
Escoffier, Jessica; Krapf, Dario; Navarrete, Felipe; Darszon, Alberto; Visconti, Pablo E.
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Mammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na+ channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na+ concentration ([Na+](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na+ concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na+](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na+](i), suggesting that also in these cells [Na+](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na+](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na+](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.
Fil: Escoffier, Jessica. University of Massachussets; Estados Unidos
Fil: Krapf, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Navarrete, Felipe. University of Massachussets; Estados Unidos
Fil: Darszon, Alberto. Universidad Nacional Autónoma de México. Instituto de Biotecnología; México
Fil: Visconti, Pablo E.. University of Massachussets; Estados Unidos
Materia
Capacitation
Flow Cytometry
ENaC
CFTR
Hyperpolarization
Membrane Potential
Sodium
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:11336/269664

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network_name_str CONICET Digital (CONICET)
spelling Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitationEscoffier, JessicaKrapf, DarioNavarrete, FelipeDarszon, AlbertoVisconti, Pablo E.CapacitationFlow CytometryENaCCFTRHyperpolarizationMembrane PotentialSodiumhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Mammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na+ channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na+ concentration ([Na+](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na+ concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na+](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na+](i), suggesting that also in these cells [Na+](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na+](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na+](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.Fil: Escoffier, Jessica. University of Massachussets; Estados UnidosFil: Krapf, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Navarrete, Felipe. University of Massachussets; Estados UnidosFil: Darszon, Alberto. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Visconti, Pablo E.. University of Massachussets; Estados UnidosCompany of Biologists2012-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/269664Escoffier, Jessica; Krapf, Dario; Navarrete, Felipe; Darszon, Alberto; Visconti, Pablo E.; Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation; Company of Biologists; Journal of Cell Science; 125; 2; 2-2012; 473-4850021-9533CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://pmc.ncbi.nlm.nih.gov/articles/PMC3283877/info:eu-repo/semantics/altIdentifier/url/https://journals.biologists.com/jcs/article/125/2/473/33173/Flow-cytometry-analysis-reveals-a-decrease-ininfo:eu-repo/semantics/altIdentifier/doi/10.1242/jcs.093344info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:46:31Zoai:ri.conicet.gov.ar:11336/269664instacron: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:34982025-09-29 10:46:31.571CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
title Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
spellingShingle Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
Escoffier, Jessica
Capacitation
Flow Cytometry
ENaC
CFTR
Hyperpolarization
Membrane Potential
Sodium
title_short Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
title_full Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
title_fullStr Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
title_full_unstemmed Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
title_sort Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation
dc.creator.none.fl_str_mv Escoffier, Jessica
Krapf, Dario
Navarrete, Felipe
Darszon, Alberto
Visconti, Pablo E.
author Escoffier, Jessica
author_facet Escoffier, Jessica
Krapf, Dario
Navarrete, Felipe
Darszon, Alberto
Visconti, Pablo E.
author_role author
author2 Krapf, Dario
Navarrete, Felipe
Darszon, Alberto
Visconti, Pablo E.
author2_role author
author
author
author
dc.subject.none.fl_str_mv Capacitation
Flow Cytometry
ENaC
CFTR
Hyperpolarization
Membrane Potential
Sodium
topic Capacitation
Flow Cytometry
ENaC
CFTR
Hyperpolarization
Membrane Potential
Sodium
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Mammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na+ channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na+ concentration ([Na+](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na+ concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na+](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na+](i), suggesting that also in these cells [Na+](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na+](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na+](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.
Fil: Escoffier, Jessica. University of Massachussets; Estados Unidos
Fil: Krapf, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Navarrete, Felipe. University of Massachussets; Estados Unidos
Fil: Darszon, Alberto. Universidad Nacional Autónoma de México. Instituto de Biotecnología; México
Fil: Visconti, Pablo E.. University of Massachussets; Estados Unidos
description Mammalian sperm require time in the female tract in order to be able to fertilize an egg. The physiological changes that render the sperm able to fertilize are known as capacitation. Capacitation is associated with an increase in intracellular pH, an increase in intracellular calcium and phosphorylation of different proteins. This process is also accompanied by the hyperpolarization of the sperm plasma membrane potential. Recently, we presented evidence showing that epithelial Na+ channels (ENaC) are present in mature sperm and that ENaCs are blocked during capacitation. In the present work, we used flow cytometry to analyze changes in intracellular Na+ concentration ([Na+](i)) during capacitation in individual cells. Our results indicate that capacitated sperm have lower Na+ concentrations. Using sperm with green fluorescent protein in their acrosomes, it was shown that the lower [Na+](i) concentration only occurs in sperm having intact acrosomes. ENaC inhibition has been shown in other cell types to depend on the activation of cystic fibrosis transmembrane conductance regulator (CFTR). In non-capacitated sperm, amiloride, an ENaC inhibitor, and genistein, a CFTR activator, caused a decrease in [Na+](i), suggesting that also in these cells [Na+](i) is dependent on the crosstalk between ENaC and CFTR. In addition, PKA inhibition blocked [Na+](i) decrease in capacitated sperm. Altogether, these data are consistent with the hypothesis that the capacitation-associated hyperpolarization involves a decrease in [Na+](i) mediated by inhibition of ENaC and regulated by PKA through activation of CFTR channels.
publishDate 2012
dc.date.none.fl_str_mv 2012-02
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/11336/269664
Escoffier, Jessica; Krapf, Dario; Navarrete, Felipe; Darszon, Alberto; Visconti, Pablo E.; Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation; Company of Biologists; Journal of Cell Science; 125; 2; 2-2012; 473-485
0021-9533
CONICET Digital
CONICET
url http://hdl.handle.net/11336/269664
identifier_str_mv Escoffier, Jessica; Krapf, Dario; Navarrete, Felipe; Darszon, Alberto; Visconti, Pablo E.; Flow cytometry analysis reveals a decrease in intracellular sodium during sperm capacitation; Company of Biologists; Journal of Cell Science; 125; 2; 2-2012; 473-485
0021-9533
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://pmc.ncbi.nlm.nih.gov/articles/PMC3283877/
info:eu-repo/semantics/altIdentifier/url/https://journals.biologists.com/jcs/article/125/2/473/33173/Flow-cytometry-analysis-reveals-a-decrease-in
info:eu-repo/semantics/altIdentifier/doi/10.1242/jcs.093344
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 Company of Biologists
publisher.none.fl_str_mv Company of Biologists
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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