Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm

Autores
Stival, Cintia Estefanía; la Spina, Florenza Antonella; Baró Graf, Carolina; Arcelay, Enid; Arranz, Silvia Eda; Ferreira, Juan J.; Le Grand, Sibylle; Dzikunu, Victor A.; Santi, Celia M.; Visconti, Pablo E.; Buffone, Mariano Gabriel; Krapf, Dario
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among signalling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have previously shown that mice deficient on the tyr-kinase cSrc are infertile, and exhibit an improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti phosphor Y416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompany capacitation. In addition, we show that cSrc inhibition also blocked the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that the capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on heterologously expressed SLO3 channel. Our results indicate that, similarly to SLO1 K+ channels, cSrc blockers decreased significantly SLO3-mediated currents. Altogether these results are consistent with findings that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.
Fil: Stival, Cintia Estefanía. 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: la Spina, Florenza Antonella. 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: Baró Graf, Carolina. 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: Arcelay, Enid. University of Massachussets; Estados Unidos
Fil: Arranz, Silvia Eda. 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: Ferreira, Juan J.. University of Washington. School of Medicine; Estados Unidos
Fil: Le Grand, Sibylle. University of Washington. School of Medicine; Estados Unidos
Fil: Dzikunu, Victor A.. University of Washington. School of Medicine; Estados Unidos
Fil: Santi, Celia M.. University of Washington. School of Medicine; Estados Unidos
Fil: Visconti, Pablo E.. University of Massachussets; Estados Unidos
Fil: Buffone, Mariano Gabriel. 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, 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
Materia
Src
Reaccion Acrosomal
Fecundacion
Espermatozoide
Pka
Membrane Potential
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:11336/23062
Acceder
id CONICETDig_29fc3876f959ed49778f462ca085f987
oai_identifier_str oai:ri.conicet.gov.ar:11336/23062
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse spermStival, Cintia Estefaníala Spina, Florenza AntonellaBaró Graf, CarolinaArcelay, EnidArranz, Silvia EdaFerreira, Juan J.Le Grand, SibylleDzikunu, Victor A.Santi, Celia M.Visconti, Pablo E.Buffone, Mariano GabrielKrapf, DarioSrcReaccion AcrosomalFecundacionEspermatozoidePkaMembrane Potentialhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among signalling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have previously shown that mice deficient on the tyr-kinase cSrc are infertile, and exhibit an improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti phosphor Y416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompany capacitation. In addition, we show that cSrc inhibition also blocked the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that the capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on heterologously expressed SLO3 channel. Our results indicate that, similarly to SLO1 K+ channels, cSrc blockers decreased significantly SLO3-mediated currents. Altogether these results are consistent with findings that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.Fil: Stival, Cintia Estefanía. 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: la Spina, Florenza Antonella. 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: Baró Graf, Carolina. 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: Arcelay, Enid. University of Massachussets; Estados UnidosFil: Arranz, Silvia Eda. 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: Ferreira, Juan J.. University of Washington. School of Medicine; Estados UnidosFil: Le Grand, Sibylle. University of Washington. School of Medicine; Estados UnidosFil: Dzikunu, Victor A.. University of Washington. School of Medicine; Estados UnidosFil: Santi, Celia M.. University of Washington. School of Medicine; Estados UnidosFil: Visconti, Pablo E.. University of Massachussets; Estados UnidosFil: Buffone, Mariano Gabriel. 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, 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; ArgentinaAmerican Society for Biochemistry and Molecular Biology2015-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/23062Stival, Cintia Estefanía; la Spina, Florenza Antonella; Baró Graf, Carolina; Arcelay, Enid; Arranz, Silvia Eda; et al.; Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 24; 30; 6-2015; 18855-188640021-92581083-351XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/290/30/18855.longinfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M115.640326info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4513139/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-23T13:43:52Zoai:ri.conicet.gov.ar:11336/23062instacron: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:34982025-12-23 13:43:52.45CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
title Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
spellingShingle Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
Stival, Cintia Estefanía
Src
Reaccion Acrosomal
Fecundacion
Espermatozoide
Pka
Membrane Potential
title_short Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
title_full Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
title_fullStr Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
title_full_unstemmed Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
title_sort Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm
dc.creator.none.fl_str_mv Stival, Cintia Estefanía
la Spina, Florenza Antonella
Baró Graf, Carolina
Arcelay, Enid
Arranz, Silvia Eda
Ferreira, Juan J.
Le Grand, Sibylle
Dzikunu, Victor A.
Santi, Celia M.
Visconti, Pablo E.
Buffone, Mariano Gabriel
Krapf, Dario
author Stival, Cintia Estefanía
author_facet Stival, Cintia Estefanía
la Spina, Florenza Antonella
Baró Graf, Carolina
Arcelay, Enid
Arranz, Silvia Eda
Ferreira, Juan J.
Le Grand, Sibylle
Dzikunu, Victor A.
Santi, Celia M.
Visconti, Pablo E.
Buffone, Mariano Gabriel
Krapf, Dario
author_role author
author2 la Spina, Florenza Antonella
Baró Graf, Carolina
Arcelay, Enid
Arranz, Silvia Eda
Ferreira, Juan J.
Le Grand, Sibylle
Dzikunu, Victor A.
Santi, Celia M.
Visconti, Pablo E.
Buffone, Mariano Gabriel
Krapf, Dario
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Src
Reaccion Acrosomal
Fecundacion
Espermatozoide
Pka
Membrane Potential
topic Src
Reaccion Acrosomal
Fecundacion
Espermatozoide
Pka
Membrane Potential
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among signalling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have previously shown that mice deficient on the tyr-kinase cSrc are infertile, and exhibit an improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti phosphor Y416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompany capacitation. In addition, we show that cSrc inhibition also blocked the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that the capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on heterologously expressed SLO3 channel. Our results indicate that, similarly to SLO1 K+ channels, cSrc blockers decreased significantly SLO3-mediated currents. Altogether these results are consistent with findings that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.
Fil: Stival, Cintia Estefanía. 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: la Spina, Florenza Antonella. 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: Baró Graf, Carolina. 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: Arcelay, Enid. University of Massachussets; Estados Unidos
Fil: Arranz, Silvia Eda. 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: Ferreira, Juan J.. University of Washington. School of Medicine; Estados Unidos
Fil: Le Grand, Sibylle. University of Washington. School of Medicine; Estados Unidos
Fil: Dzikunu, Victor A.. University of Washington. School of Medicine; Estados Unidos
Fil: Santi, Celia M.. University of Washington. School of Medicine; Estados Unidos
Fil: Visconti, Pablo E.. University of Massachussets; Estados Unidos
Fil: Buffone, Mariano Gabriel. 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, 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
description Plasma membrane hyperpolarization is crucial for mammalian sperm to acquire acrosomal responsiveness during capacitation. Among signalling events leading to mammalian sperm capacitation, the immediate activation of protein kinase A plays a pivotal role, promoting the subsequent stimulation of protein tyrosine phosphorylation that associates with fertilizing capacity. We have previously shown that mice deficient on the tyr-kinase cSrc are infertile, and exhibit an improper cauda epididymis development. It is therefore not clear whether lack of sperm functionality is due to problems in epididymal maturation or to the absence of cSrc in sperm. To further address this problem, we investigated the kinetics of cSrc activation using anti phosphor Y416-cSrc antibodies that only recognize active cSrc. Our results provide evidence that cSrc is activated downstream of PKA and that inhibition of its activity blocks the capacitation-induced hyperpolarization of the sperm plasma membrane without blocking the increase in tyrosine phosphorylation that accompany capacitation. In addition, we show that cSrc inhibition also blocked the agonist-induced acrosome reaction and that this inhibition is overcome by pharmacological hyperpolarization. Considering that the capacitation-induced hyperpolarization is mediated by SLO3, we evaluated the action of cSrc inhibitors on heterologously expressed SLO3 channel. Our results indicate that, similarly to SLO1 K+ channels, cSrc blockers decreased significantly SLO3-mediated currents. Altogether these results are consistent with findings that hyperpolarization of the sperm plasma membrane is necessary and sufficient to prepare the sperm for the acrosome reaction and suggest that changes in sperm membrane potential are mediated by cSrc activation.
publishDate 2015
dc.date.none.fl_str_mv 2015-06
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/11336/23062
Stival, Cintia Estefanía; la Spina, Florenza Antonella; Baró Graf, Carolina; Arcelay, Enid; Arranz, Silvia Eda; et al.; Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 24; 30; 6-2015; 18855-18864
0021-9258
1083-351X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/23062
identifier_str_mv Stival, Cintia Estefanía; la Spina, Florenza Antonella; Baró Graf, Carolina; Arcelay, Enid; Arranz, Silvia Eda; et al.; Src is the connecting player between PKA activation and hyperpolarization through SLO3 regulation in mouse sperm; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 24; 30; 6-2015; 18855-18864
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/url/http://www.jbc.org/content/290/30/18855.long
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M115.640326
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4513139/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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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)
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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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