Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa

Autores
Laiz Quiroga, Lucia; Navarro, Micaela; Martínez León, Eduardo Antonio; Buffone, Mariano Gabriel; Mutto, Adrián Angel; Osycka Salut, Claudia Elena
Año de publicación
2023
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Spermatozoa must undergo three events to fertilize anoocyte in vivo: capacitation, hyperactivation and acrosome reaction. Therefore,it is necessary to induce these physiological events in vitro in themasculine gamete during assisted reproductive techniques. Even though the invitro conditions under which many species acquire their fertilizing abilityare well known, to this day, there is no standard protocol to induce theseevents in equine cryopreserved spermatozoa. Thus, there is no available protocolfor embryo production through in vitro fertilization (IVF) with these samples.It has been proven that brief exposure of murine andbovine spermatozoa to calcium ionophore A23187 and the consequent calcium influxto the cell cytoplasm increases their fertilizing ability in vitro andembryo production due to the induction of capacitation and hyperactivation. Moreover,A23187 increases events related to capacitation without inducing the classicalcapacitation signaling pathway such as PKA-activation (pPKA) or proteintyrosine phosphorylation (pTyr). Given the current difficulties in equine embryoproduction through IVF with cryopreserved spermatozoa and the previously showneffects of the calcium ionophore in spermatozoa from other species, this workaimed to study the potential fertilizing ability of equine cryopreserved spermatozoathat were previously exposed to A23187.First, we studied the effect of A23187 (1 µM) on spermatozoa.After 10 minutes of exposure, the ionophore decreased the spermatozoa motility(CASA, p<0,05) without affecting their viability (HOS test, p>0,05) or acrosomestatus (PSA-FITC, IF, p>0,05). After removing the ionophore, the spermatozoawere incubated under non-capacitating conditions for 20 minutes and motilitywas reassessed, and their fertilizing ability was studied. The previousincubation with A23187 increased the hyperactivated sperm population (CASA,p<0,05) and the induction of capacitation-associated events such as progesterone-inducedacrosome reaction (IF, p<0,05) and the sperm ability to bind to bovineoocyte zona pellucida (p<0,001) without activating the pTyr and pPKA pathways(IF, p>0,05). Our results suggest that brief exposure of cryopreservedequine spermatozoa to calcium ionophore A23187 could be incorporated into the assistedreproductive techniques to increase spermatozoa fertilizing ability in vitrodue to its effect on hyperactivation and capacitation in this species.
Fil: Laiz Quiroga, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Navarro, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Martínez León, Eduardo Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
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: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Osycka Salut, Claudia Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
XXIV Jornadas Anuales de la Sociedad Argentina de Biología
Ciudad de Buenos Aires
Argentina
Sociedad Argentina de Biología
Instituto de Biología y Medicina Experimental
Sociedad Uruguaya de Biociencias
Materia
EQUINE SPERMATOZOA
CAPACITATION
CALCIUM IONOPHORE
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:11336/247604
Acceder
id CONICETDig_d43ebe5c4075f449a667e195427b6d96
oai_identifier_str oai:ri.conicet.gov.ar:11336/247604
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoaLaiz Quiroga, LuciaNavarro, MicaelaMartínez León, Eduardo AntonioBuffone, Mariano GabrielMutto, Adrián AngelOsycka Salut, Claudia ElenaEQUINE SPERMATOZOACAPACITATIONCALCIUM IONOPHOREhttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Spermatozoa must undergo three events to fertilize anoocyte in vivo: capacitation, hyperactivation and acrosome reaction. Therefore,it is necessary to induce these physiological events in vitro in themasculine gamete during assisted reproductive techniques. Even though the invitro conditions under which many species acquire their fertilizing abilityare well known, to this day, there is no standard protocol to induce theseevents in equine cryopreserved spermatozoa. Thus, there is no available protocolfor embryo production through in vitro fertilization (IVF) with these samples.It has been proven that brief exposure of murine andbovine spermatozoa to calcium ionophore A23187 and the consequent calcium influxto the cell cytoplasm increases their fertilizing ability in vitro andembryo production due to the induction of capacitation and hyperactivation. Moreover,A23187 increases events related to capacitation without inducing the classicalcapacitation signaling pathway such as PKA-activation (pPKA) or proteintyrosine phosphorylation (pTyr). Given the current difficulties in equine embryoproduction through IVF with cryopreserved spermatozoa and the previously showneffects of the calcium ionophore in spermatozoa from other species, this workaimed to study the potential fertilizing ability of equine cryopreserved spermatozoathat were previously exposed to A23187.First, we studied the effect of A23187 (1 µM) on spermatozoa.After 10 minutes of exposure, the ionophore decreased the spermatozoa motility(CASA, p<0,05) without affecting their viability (HOS test, p>0,05) or acrosomestatus (PSA-FITC, IF, p>0,05). After removing the ionophore, the spermatozoawere incubated under non-capacitating conditions for 20 minutes and motilitywas reassessed, and their fertilizing ability was studied. The previousincubation with A23187 increased the hyperactivated sperm population (CASA,p<0,05) and the induction of capacitation-associated events such as progesterone-inducedacrosome reaction (IF, p<0,05) and the sperm ability to bind to bovineoocyte zona pellucida (p<0,001) without activating the pTyr and pPKA pathways(IF, p>0,05). Our results suggest that brief exposure of cryopreservedequine spermatozoa to calcium ionophore A23187 could be incorporated into the assistedreproductive techniques to increase spermatozoa fertilizing ability in vitrodue to its effect on hyperactivation and capacitation in this species.Fil: Laiz Quiroga, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Navarro, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Martínez León, Eduardo Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; ArgentinaFil: 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: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Osycka Salut, Claudia Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaXXIV Jornadas Anuales de la Sociedad Argentina de BiologíaCiudad de Buenos AiresArgentinaSociedad Argentina de BiologíaInstituto de Biología y Medicina ExperimentalSociedad Uruguaya de BiocienciasSociedad Argentina de Biología2023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectJornadaJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/247604Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa; XXIV Jornadas Anuales de la Sociedad Argentina de Biología; Ciudad de Buenos Aires; Argentina; 2022; 1-31667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biologia.org.ar/jornadas-anteriores/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:07:17Zoai:ri.conicet.gov.ar:11336/247604instacron: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 13:07:17.539CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
title Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
spellingShingle Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
Laiz Quiroga, Lucia
EQUINE SPERMATOZOA
CAPACITATION
CALCIUM IONOPHORE
title_short Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
title_full Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
title_fullStr Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
title_full_unstemmed Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
title_sort Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa
dc.creator.none.fl_str_mv Laiz Quiroga, Lucia
Navarro, Micaela
Martínez León, Eduardo Antonio
Buffone, Mariano Gabriel
Mutto, Adrián Angel
Osycka Salut, Claudia Elena
author Laiz Quiroga, Lucia
author_facet Laiz Quiroga, Lucia
Navarro, Micaela
Martínez León, Eduardo Antonio
Buffone, Mariano Gabriel
Mutto, Adrián Angel
Osycka Salut, Claudia Elena
author_role author
author2 Navarro, Micaela
Martínez León, Eduardo Antonio
Buffone, Mariano Gabriel
Mutto, Adrián Angel
Osycka Salut, Claudia Elena
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv EQUINE SPERMATOZOA
CAPACITATION
CALCIUM IONOPHORE
topic EQUINE SPERMATOZOA
CAPACITATION
CALCIUM IONOPHORE
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Spermatozoa must undergo three events to fertilize anoocyte in vivo: capacitation, hyperactivation and acrosome reaction. Therefore,it is necessary to induce these physiological events in vitro in themasculine gamete during assisted reproductive techniques. Even though the invitro conditions under which many species acquire their fertilizing abilityare well known, to this day, there is no standard protocol to induce theseevents in equine cryopreserved spermatozoa. Thus, there is no available protocolfor embryo production through in vitro fertilization (IVF) with these samples.It has been proven that brief exposure of murine andbovine spermatozoa to calcium ionophore A23187 and the consequent calcium influxto the cell cytoplasm increases their fertilizing ability in vitro andembryo production due to the induction of capacitation and hyperactivation. Moreover,A23187 increases events related to capacitation without inducing the classicalcapacitation signaling pathway such as PKA-activation (pPKA) or proteintyrosine phosphorylation (pTyr). Given the current difficulties in equine embryoproduction through IVF with cryopreserved spermatozoa and the previously showneffects of the calcium ionophore in spermatozoa from other species, this workaimed to study the potential fertilizing ability of equine cryopreserved spermatozoathat were previously exposed to A23187.First, we studied the effect of A23187 (1 µM) on spermatozoa.After 10 minutes of exposure, the ionophore decreased the spermatozoa motility(CASA, p<0,05) without affecting their viability (HOS test, p>0,05) or acrosomestatus (PSA-FITC, IF, p>0,05). After removing the ionophore, the spermatozoawere incubated under non-capacitating conditions for 20 minutes and motilitywas reassessed, and their fertilizing ability was studied. The previousincubation with A23187 increased the hyperactivated sperm population (CASA,p<0,05) and the induction of capacitation-associated events such as progesterone-inducedacrosome reaction (IF, p<0,05) and the sperm ability to bind to bovineoocyte zona pellucida (p<0,001) without activating the pTyr and pPKA pathways(IF, p>0,05). Our results suggest that brief exposure of cryopreservedequine spermatozoa to calcium ionophore A23187 could be incorporated into the assistedreproductive techniques to increase spermatozoa fertilizing ability in vitrodue to its effect on hyperactivation and capacitation in this species.
Fil: Laiz Quiroga, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Navarro, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Martínez León, Eduardo Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina
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: Mutto, Adrián Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Osycka Salut, Claudia Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
XXIV Jornadas Anuales de la Sociedad Argentina de Biología
Ciudad de Buenos Aires
Argentina
Sociedad Argentina de Biología
Instituto de Biología y Medicina Experimental
Sociedad Uruguaya de Biociencias
description Spermatozoa must undergo three events to fertilize anoocyte in vivo: capacitation, hyperactivation and acrosome reaction. Therefore,it is necessary to induce these physiological events in vitro in themasculine gamete during assisted reproductive techniques. Even though the invitro conditions under which many species acquire their fertilizing abilityare well known, to this day, there is no standard protocol to induce theseevents in equine cryopreserved spermatozoa. Thus, there is no available protocolfor embryo production through in vitro fertilization (IVF) with these samples.It has been proven that brief exposure of murine andbovine spermatozoa to calcium ionophore A23187 and the consequent calcium influxto the cell cytoplasm increases their fertilizing ability in vitro andembryo production due to the induction of capacitation and hyperactivation. Moreover,A23187 increases events related to capacitation without inducing the classicalcapacitation signaling pathway such as PKA-activation (pPKA) or proteintyrosine phosphorylation (pTyr). Given the current difficulties in equine embryoproduction through IVF with cryopreserved spermatozoa and the previously showneffects of the calcium ionophore in spermatozoa from other species, this workaimed to study the potential fertilizing ability of equine cryopreserved spermatozoathat were previously exposed to A23187.First, we studied the effect of A23187 (1 µM) on spermatozoa.After 10 minutes of exposure, the ionophore decreased the spermatozoa motility(CASA, p<0,05) without affecting their viability (HOS test, p>0,05) or acrosomestatus (PSA-FITC, IF, p>0,05). After removing the ionophore, the spermatozoawere incubated under non-capacitating conditions for 20 minutes and motilitywas reassessed, and their fertilizing ability was studied. The previousincubation with A23187 increased the hyperactivated sperm population (CASA,p<0,05) and the induction of capacitation-associated events such as progesterone-inducedacrosome reaction (IF, p<0,05) and the sperm ability to bind to bovineoocyte zona pellucida (p<0,001) without activating the pTyr and pPKA pathways(IF, p>0,05). Our results suggest that brief exposure of cryopreservedequine spermatozoa to calcium ionophore A23187 could be incorporated into the assistedreproductive techniques to increase spermatozoa fertilizing ability in vitrodue to its effect on hyperactivation and capacitation in this species.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Jornada
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/11336/247604
Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa; XXIV Jornadas Anuales de la Sociedad Argentina de Biología; Ciudad de Buenos Aires; Argentina; 2022; 1-3
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/247604
identifier_str_mv Calcium ionophore a23187 effect on acquisition of fertilizing ability in equine cryopreserved spermatozoa; XXIV Jornadas Anuales de la Sociedad Argentina de Biología; Ciudad de Buenos Aires; Argentina; 2022; 1-3
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Biología
publisher.none.fl_str_mv Sociedad Argentina de Biología
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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