Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa

Autores
Diana, A.; Buttazzoni, A.; Sosa, M. A.; Vieira, L.; Aguilera, Andrea Carolina
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
In mammals, only a small number of ejaculated spermatozoa (SPZ) reach the region of the oviduct (ampulla) after the copula, where they encounterand fertilize the egg. It has been suggested that a sperm subpopulation is selected during their transit trough the female genital tract, so that only thosewith high fertilizing capability and the best skills for supporting embryo development can fertilize the egg. Fluids of the female genital tract, such asthe follicular (FF), the oviductal fluid (OF), and the secretion of cumulus-oocyte complex (COCs), could promote the SPZ chemotaxis to thefertilization site. Progesterone (P4) is considered as an effective chemoattractant in most mammalian species, though other components of the fluidscould attract SPZ even more efficiently. In the present study, we evaluated possible changes in carbohydrate composition of sperm surface aftercapacitation and chemotaxis. For this, we determined the content of N-acetyl-glucosamine (NAG) in porcine SPZ after the mentioned processes byusing WGA-FITC lectin and flow cytometry. We observed that NAG content was significantly higher in the capacitated SPZ (30 min in capacitationmedia TALP, at 38.5ºC and with 5% CO2) compared to fresh SPZ or SPZ stored in BTS (diluent media). For the chemotaxis assays, OF and FFcollected from prepubertal gilts (OF0 and FF0) and periovulatory phase (OF2 and FF2) were used as chemoattractants. Six wells were filled withfresh spermatozoa (20×106/mL) from fertile boars (N = 3) selected in a discontinuous percoll gradient and immediately transferred to TALP,previously equilibrated at 38.5ºC and 5% CO2. The opposite wells of the chemotaxis chamber (six) were filled with TALP (control group) or TALPsupplemented with the chemoattractants as indicated: (1) TALP (control), (2) FF0 (1.25%), (3) FF2 (1.25%), (4) OF0 (1.25%), (5) OF2 (1.25%), (6)P4 (28.3 pM). After 20 min at 38.5ºC and 5% CO2, the SPZ from the opposite wells were rescued, processed for NAG detection and analyzed byflow cytometry. We observed that NAG content was significantly lower in the SPZ obtained from the groups 3 and 6 compared to the control groupor the original SPZ (P < 0.05). These preliminary results suggest that FF2 and P4 can selectively attract a SPZ subpopulation with low content ofNAG in the plasma membrane under the in-vitro conditions.
Fil: Diana, A.. Universidad de Murcia; España
Fil: Buttazzoni, A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, M. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Vieira, L.. Universidad de Murcia; España
Fil: Aguilera, Andrea Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
IV Joint Meeting of the Biology Societies of Argentina
Mendoza
Argentina
Sociedad de Biología de Cuyo
Materia
N-ACETYL-GLUCOSAMINE
SPERMATOZOA
PORCINE
CAPACITATION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/178072

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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoaDiana, A.Buttazzoni, A.Sosa, M. A.Vieira, L.Aguilera, Andrea CarolinaN-ACETYL-GLUCOSAMINESPERMATOZOAPORCINECAPACITATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In mammals, only a small number of ejaculated spermatozoa (SPZ) reach the region of the oviduct (ampulla) after the copula, where they encounterand fertilize the egg. It has been suggested that a sperm subpopulation is selected during their transit trough the female genital tract, so that only thosewith high fertilizing capability and the best skills for supporting embryo development can fertilize the egg. Fluids of the female genital tract, such asthe follicular (FF), the oviductal fluid (OF), and the secretion of cumulus-oocyte complex (COCs), could promote the SPZ chemotaxis to thefertilization site. Progesterone (P4) is considered as an effective chemoattractant in most mammalian species, though other components of the fluidscould attract SPZ even more efficiently. In the present study, we evaluated possible changes in carbohydrate composition of sperm surface aftercapacitation and chemotaxis. For this, we determined the content of N-acetyl-glucosamine (NAG) in porcine SPZ after the mentioned processes byusing WGA-FITC lectin and flow cytometry. We observed that NAG content was significantly higher in the capacitated SPZ (30 min in capacitationmedia TALP, at 38.5ºC and with 5% CO2) compared to fresh SPZ or SPZ stored in BTS (diluent media). For the chemotaxis assays, OF and FFcollected from prepubertal gilts (OF0 and FF0) and periovulatory phase (OF2 and FF2) were used as chemoattractants. Six wells were filled withfresh spermatozoa (20×106/mL) from fertile boars (N = 3) selected in a discontinuous percoll gradient and immediately transferred to TALP,previously equilibrated at 38.5ºC and 5% CO2. The opposite wells of the chemotaxis chamber (six) were filled with TALP (control group) or TALPsupplemented with the chemoattractants as indicated: (1) TALP (control), (2) FF0 (1.25%), (3) FF2 (1.25%), (4) OF0 (1.25%), (5) OF2 (1.25%), (6)P4 (28.3 pM). After 20 min at 38.5ºC and 5% CO2, the SPZ from the opposite wells were rescued, processed for NAG detection and analyzed byflow cytometry. We observed that NAG content was significantly lower in the SPZ obtained from the groups 3 and 6 compared to the control groupor the original SPZ (P < 0.05). These preliminary results suggest that FF2 and P4 can selectively attract a SPZ subpopulation with low content ofNAG in the plasma membrane under the in-vitro conditions.Fil: Diana, A.. Universidad de Murcia; EspañaFil: Buttazzoni, A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Sosa, M. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Vieira, L.. Universidad de Murcia; EspañaFil: Aguilera, Andrea Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaIV Joint Meeting of the Biology Societies of ArgentinaMendozaArgentinaSociedad de Biología de CuyoTech Science Press2021info: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/11336/178072Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa; IV Joint Meeting of the Biology Societies of Argentina; Mendoza; Argentina; 2020; 28-280327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://sbcuyo.org.ar/wp-content/uploads/2021/08/BIOCELL-Vol_-45-Suppl_-3-2021.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:10:59Zoai:ri.conicet.gov.ar:11336/178072instacron: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:34982025-09-03 10:11:00.046CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
title Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
spellingShingle Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
Diana, A.
N-ACETYL-GLUCOSAMINE
SPERMATOZOA
PORCINE
CAPACITATION
title_short Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
title_full Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
title_fullStr Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
title_full_unstemmed Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
title_sort Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa
dc.creator.none.fl_str_mv Diana, A.
Buttazzoni, A.
Sosa, M. A.
Vieira, L.
Aguilera, Andrea Carolina
author Diana, A.
author_facet Diana, A.
Buttazzoni, A.
Sosa, M. A.
Vieira, L.
Aguilera, Andrea Carolina
author_role author
author2 Buttazzoni, A.
Sosa, M. A.
Vieira, L.
Aguilera, Andrea Carolina
author2_role author
author
author
author
dc.subject.none.fl_str_mv N-ACETYL-GLUCOSAMINE
SPERMATOZOA
PORCINE
CAPACITATION
topic N-ACETYL-GLUCOSAMINE
SPERMATOZOA
PORCINE
CAPACITATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In mammals, only a small number of ejaculated spermatozoa (SPZ) reach the region of the oviduct (ampulla) after the copula, where they encounterand fertilize the egg. It has been suggested that a sperm subpopulation is selected during their transit trough the female genital tract, so that only thosewith high fertilizing capability and the best skills for supporting embryo development can fertilize the egg. Fluids of the female genital tract, such asthe follicular (FF), the oviductal fluid (OF), and the secretion of cumulus-oocyte complex (COCs), could promote the SPZ chemotaxis to thefertilization site. Progesterone (P4) is considered as an effective chemoattractant in most mammalian species, though other components of the fluidscould attract SPZ even more efficiently. In the present study, we evaluated possible changes in carbohydrate composition of sperm surface aftercapacitation and chemotaxis. For this, we determined the content of N-acetyl-glucosamine (NAG) in porcine SPZ after the mentioned processes byusing WGA-FITC lectin and flow cytometry. We observed that NAG content was significantly higher in the capacitated SPZ (30 min in capacitationmedia TALP, at 38.5ºC and with 5% CO2) compared to fresh SPZ or SPZ stored in BTS (diluent media). For the chemotaxis assays, OF and FFcollected from prepubertal gilts (OF0 and FF0) and periovulatory phase (OF2 and FF2) were used as chemoattractants. Six wells were filled withfresh spermatozoa (20×106/mL) from fertile boars (N = 3) selected in a discontinuous percoll gradient and immediately transferred to TALP,previously equilibrated at 38.5ºC and 5% CO2. The opposite wells of the chemotaxis chamber (six) were filled with TALP (control group) or TALPsupplemented with the chemoattractants as indicated: (1) TALP (control), (2) FF0 (1.25%), (3) FF2 (1.25%), (4) OF0 (1.25%), (5) OF2 (1.25%), (6)P4 (28.3 pM). After 20 min at 38.5ºC and 5% CO2, the SPZ from the opposite wells were rescued, processed for NAG detection and analyzed byflow cytometry. We observed that NAG content was significantly lower in the SPZ obtained from the groups 3 and 6 compared to the control groupor the original SPZ (P < 0.05). These preliminary results suggest that FF2 and P4 can selectively attract a SPZ subpopulation with low content ofNAG in the plasma membrane under the in-vitro conditions.
Fil: Diana, A.. Universidad de Murcia; España
Fil: Buttazzoni, A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Sosa, M. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Vieira, L.. Universidad de Murcia; España
Fil: Aguilera, Andrea Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
IV Joint Meeting of the Biology Societies of Argentina
Mendoza
Argentina
Sociedad de Biología de Cuyo
description In mammals, only a small number of ejaculated spermatozoa (SPZ) reach the region of the oviduct (ampulla) after the copula, where they encounterand fertilize the egg. It has been suggested that a sperm subpopulation is selected during their transit trough the female genital tract, so that only thosewith high fertilizing capability and the best skills for supporting embryo development can fertilize the egg. Fluids of the female genital tract, such asthe follicular (FF), the oviductal fluid (OF), and the secretion of cumulus-oocyte complex (COCs), could promote the SPZ chemotaxis to thefertilization site. Progesterone (P4) is considered as an effective chemoattractant in most mammalian species, though other components of the fluidscould attract SPZ even more efficiently. In the present study, we evaluated possible changes in carbohydrate composition of sperm surface aftercapacitation and chemotaxis. For this, we determined the content of N-acetyl-glucosamine (NAG) in porcine SPZ after the mentioned processes byusing WGA-FITC lectin and flow cytometry. We observed that NAG content was significantly higher in the capacitated SPZ (30 min in capacitationmedia TALP, at 38.5ºC and with 5% CO2) compared to fresh SPZ or SPZ stored in BTS (diluent media). For the chemotaxis assays, OF and FFcollected from prepubertal gilts (OF0 and FF0) and periovulatory phase (OF2 and FF2) were used as chemoattractants. Six wells were filled withfresh spermatozoa (20×106/mL) from fertile boars (N = 3) selected in a discontinuous percoll gradient and immediately transferred to TALP,previously equilibrated at 38.5ºC and 5% CO2. The opposite wells of the chemotaxis chamber (six) were filled with TALP (control group) or TALPsupplemented with the chemoattractants as indicated: (1) TALP (control), (2) FF0 (1.25%), (3) FF2 (1.25%), (4) OF0 (1.25%), (5) OF2 (1.25%), (6)P4 (28.3 pM). After 20 min at 38.5ºC and 5% CO2, the SPZ from the opposite wells were rescued, processed for NAG detection and analyzed byflow cytometry. We observed that NAG content was significantly lower in the SPZ obtained from the groups 3 and 6 compared to the control groupor the original SPZ (P < 0.05). These preliminary results suggest that FF2 and P4 can selectively attract a SPZ subpopulation with low content ofNAG in the plasma membrane under the in-vitro conditions.
publishDate 2021
dc.date.none.fl_str_mv 2021
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/11336/178072
Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa; IV Joint Meeting of the Biology Societies of Argentina; Mendoza; Argentina; 2020; 28-28
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/178072
identifier_str_mv Changes in n-acetyglucosamine content associated with capacitation and chemotaix in porcine spermatozoa; IV Joint Meeting of the Biology Societies of Argentina; Mendoza; Argentina; 2020; 28-28
0327-9545
1667-5746
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://sbcuyo.org.ar/wp-content/uploads/2021/08/BIOCELL-Vol_-45-Suppl_-3-2021.pdf
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https://creativecommons.org/licenses/by/2.5/ar/
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Tech Science Press
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dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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