Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm

Autores
Balestrini, Paula Ania; Jabloñski, Martina; Gilio, Nicolas; Luque, Guillermina Maria; Krapf, Dari; Mariano Gabriel Buffone
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
To gain fertilization ability sperm must undergo a series of physiological modification inthe female tract called capacitation, which leads to the acquisition of hyperactivatedmotility (HA) and the ability to undergo acrosome reaction (AR). Membrane potential(Em) hyperpolarization and alkalinization of the intracellular pH are necessary for HAand AR to take place. It has been reported either pharmacologically or genetically(potassium channel Slo3 knockout mice) that hyperpolarization is necessary andsufficient for sperm acrosomal responsiveness. In addition, intracellular alkalinizationallows the opening of CatSper channels which lead to an increase in intracellular calciumthat is fundamental for the HA of sperm. The aim of this work was to elucidate theinterplay between Em and pH in mouse sperm. We used BCECF-AM and DISC3(5), probesthat measures changes in cytoplasmic pH and Em, respectively. First, we studied thedependence of pH and Em, by co-incubating sperm with both probes followed by FlowCytometry analysis. We observed that all the cells were grouped into two populations:1) low pH and depolarized Em or 2) high pH and hyperpolarized Em. We also determinedthat sperm incubated under non-capacitating conditions with the addition of 1 μMvalinomycin displayed a robust increase in pH suggesting that changes in Em arenecessary for the rise in pH observed during capacitation. This increase was abrogatedin the presence of high K+ in the extracellular space. In addition, we studied the kineticsof this changes by assessing Em and pH at different time points of capacitation (0 to 60min). Our results showed that there is a rapid pH alkalinization during the first 15 min ofincubation in capacitating medium that is followed by a transient acidification at 30 min.Alkalinization is then restored at 45 min and remain unchanged. Thus, our resultssuggested that changes in the Em induces modifications in intracellular pH during mousesperm capacitation.
Fil: Balestrini, Paula Ania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Jabloñski, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Gilio, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Luque, Guillermina Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Krapf, Dari. 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: Mariano Gabriel Buffone. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Asociación Argentina de Nanomedicinas
Materia
SPERM
CAPACITATION
HYPERPOLARIZATION
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:11336/132952
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/132952
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse spermBalestrini, Paula AniaJabloñski, MartinaGilio, NicolasLuque, Guillermina MariaKrapf, DariMariano Gabriel BuffoneSPERMCAPACITATIONHYPERPOLARIZATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1To gain fertilization ability sperm must undergo a series of physiological modification inthe female tract called capacitation, which leads to the acquisition of hyperactivatedmotility (HA) and the ability to undergo acrosome reaction (AR). Membrane potential(Em) hyperpolarization and alkalinization of the intracellular pH are necessary for HAand AR to take place. It has been reported either pharmacologically or genetically(potassium channel Slo3 knockout mice) that hyperpolarization is necessary andsufficient for sperm acrosomal responsiveness. In addition, intracellular alkalinizationallows the opening of CatSper channels which lead to an increase in intracellular calciumthat is fundamental for the HA of sperm. The aim of this work was to elucidate theinterplay between Em and pH in mouse sperm. We used BCECF-AM and DISC3(5), probesthat measures changes in cytoplasmic pH and Em, respectively. First, we studied thedependence of pH and Em, by co-incubating sperm with both probes followed by FlowCytometry analysis. We observed that all the cells were grouped into two populations:1) low pH and depolarized Em or 2) high pH and hyperpolarized Em. We also determinedthat sperm incubated under non-capacitating conditions with the addition of 1 μMvalinomycin displayed a robust increase in pH suggesting that changes in Em arenecessary for the rise in pH observed during capacitation. This increase was abrogatedin the presence of high K+ in the extracellular space. In addition, we studied the kineticsof this changes by assessing Em and pH at different time points of capacitation (0 to 60min). Our results showed that there is a rapid pH alkalinization during the first 15 min ofincubation in capacitating medium that is followed by a transient acidification at 30 min.Alkalinization is then restored at 45 min and remain unchanged. Thus, our resultssuggested that changes in the Em induces modifications in intracellular pH during mousesperm capacitation.Fil: Balestrini, Paula Ania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Jabloñski, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Gilio, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Luque, Guillermina Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Krapf, Dari. 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: Mariano Gabriel Buffone. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaLXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de BiologíaSociedad Argentina de ProtozoologíaAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioAsociación Argentina de NanomedicinasFundación Revista Medicina2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/132952Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio ; Mar del Plata; Argentina; 2019; 79-790025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.medicinabuenosaires.com/indices-de-2010-a-2019/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:42:23Zoai:ri.conicet.gov.ar:11336/132952instacron: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 10:42:24.071CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
title Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
spellingShingle Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
Balestrini, Paula Ania
SPERM
CAPACITATION
HYPERPOLARIZATION
title_short Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
title_full Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
title_fullStr Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
title_full_unstemmed Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
title_sort Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm
dc.creator.none.fl_str_mv Balestrini, Paula Ania
Jabloñski, Martina
Gilio, Nicolas
Luque, Guillermina Maria
Krapf, Dari
Mariano Gabriel Buffone
author Balestrini, Paula Ania
author_facet Balestrini, Paula Ania
Jabloñski, Martina
Gilio, Nicolas
Luque, Guillermina Maria
Krapf, Dari
Mariano Gabriel Buffone
author_role author
author2 Jabloñski, Martina
Gilio, Nicolas
Luque, Guillermina Maria
Krapf, Dari
Mariano Gabriel Buffone
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv SPERM
CAPACITATION
HYPERPOLARIZATION
topic SPERM
CAPACITATION
HYPERPOLARIZATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv To gain fertilization ability sperm must undergo a series of physiological modification inthe female tract called capacitation, which leads to the acquisition of hyperactivatedmotility (HA) and the ability to undergo acrosome reaction (AR). Membrane potential(Em) hyperpolarization and alkalinization of the intracellular pH are necessary for HAand AR to take place. It has been reported either pharmacologically or genetically(potassium channel Slo3 knockout mice) that hyperpolarization is necessary andsufficient for sperm acrosomal responsiveness. In addition, intracellular alkalinizationallows the opening of CatSper channels which lead to an increase in intracellular calciumthat is fundamental for the HA of sperm. The aim of this work was to elucidate theinterplay between Em and pH in mouse sperm. We used BCECF-AM and DISC3(5), probesthat measures changes in cytoplasmic pH and Em, respectively. First, we studied thedependence of pH and Em, by co-incubating sperm with both probes followed by FlowCytometry analysis. We observed that all the cells were grouped into two populations:1) low pH and depolarized Em or 2) high pH and hyperpolarized Em. We also determinedthat sperm incubated under non-capacitating conditions with the addition of 1 μMvalinomycin displayed a robust increase in pH suggesting that changes in Em arenecessary for the rise in pH observed during capacitation. This increase was abrogatedin the presence of high K+ in the extracellular space. In addition, we studied the kineticsof this changes by assessing Em and pH at different time points of capacitation (0 to 60min). Our results showed that there is a rapid pH alkalinization during the first 15 min ofincubation in capacitating medium that is followed by a transient acidification at 30 min.Alkalinization is then restored at 45 min and remain unchanged. Thus, our resultssuggested that changes in the Em induces modifications in intracellular pH during mousesperm capacitation.
Fil: Balestrini, Paula Ania. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Jabloñski, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Gilio, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Luque, Guillermina Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Krapf, Dari. 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: Mariano Gabriel Buffone. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Asociación Argentina de Nanomedicinas
description To gain fertilization ability sperm must undergo a series of physiological modification inthe female tract called capacitation, which leads to the acquisition of hyperactivatedmotility (HA) and the ability to undergo acrosome reaction (AR). Membrane potential(Em) hyperpolarization and alkalinization of the intracellular pH are necessary for HAand AR to take place. It has been reported either pharmacologically or genetically(potassium channel Slo3 knockout mice) that hyperpolarization is necessary andsufficient for sperm acrosomal responsiveness. In addition, intracellular alkalinizationallows the opening of CatSper channels which lead to an increase in intracellular calciumthat is fundamental for the HA of sperm. The aim of this work was to elucidate theinterplay between Em and pH in mouse sperm. We used BCECF-AM and DISC3(5), probesthat measures changes in cytoplasmic pH and Em, respectively. First, we studied thedependence of pH and Em, by co-incubating sperm with both probes followed by FlowCytometry analysis. We observed that all the cells were grouped into two populations:1) low pH and depolarized Em or 2) high pH and hyperpolarized Em. We also determinedthat sperm incubated under non-capacitating conditions with the addition of 1 μMvalinomycin displayed a robust increase in pH suggesting that changes in Em arenecessary for the rise in pH observed during capacitation. This increase was abrogatedin the presence of high K+ in the extracellular space. In addition, we studied the kineticsof this changes by assessing Em and pH at different time points of capacitation (0 to 60min). Our results showed that there is a rapid pH alkalinization during the first 15 min ofincubation in capacitating medium that is followed by a transient acidification at 30 min.Alkalinization is then restored at 45 min and remain unchanged. Thus, our resultssuggested that changes in the Em induces modifications in intracellular pH during mousesperm capacitation.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/132952
Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio ; Mar del Plata; Argentina; 2019; 79-79
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/132952
identifier_str_mv Membrane potential hyperpolarization induces cytoplasmic alkalinization of mouse sperm; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio ; Mar del Plata; Argentina; 2019; 79-79
0025-7680
1669-9106
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://www.medicinabuenosaires.com/indices-de-2010-a-2019/
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
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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 13.070432

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