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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/269664
Ver los metadatos del registro completo
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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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1844614507096178688 |
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13.070432 |