Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation

Autores
Roldan Olarte, Eugenia Mariela; Maillo, Verónica; Sánchez Calabuig, María Jesús; Beltrán Breña, Paula; Rizos, Dimitrios; Gutiérrez Adan, Alfonso
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
This study examines the impacts of the urokinase-type plasminogen activator (uPA) on the in vitro maturation (IVM) of bovine oocytes. Cumulus–oocyte complexes in IVM medium were treated with uPA, amiloride (an uPA inhibitor), dimethyl sulfoxide (DMSO) or left untreated (control group). After 24 h of IVM, oocytes were recovered for testing or were in vitro fertilized and cultured to the blastocyst stage. The factors examined in all groups were: (i) oocyte nuclear maturation (Hoëscht staining); (ii) oocyte cytoplasmic maturation (cortical granules, CGs, distribution assessed by LCA-FITC); (iii) oocyte and cumulus cell (CC) gene expression (RT-qPCR); and (iv) embryo development (cleavage rate and blastocyst yield). Oocytes subjected to uPA treatment showed rates of nuclear maturation and CG distribution patterns similar to controls (P> 0.05), whereas lower rates of oocyte maturation were recorded in the amiloride group (P< 0.05). Both in oocytes and CC, treatment with uPA did not affect the transcription of genes related to apoptosis, cell junctions, cell cycle or serpin protease inhibitors. In contrast, amiloride altered the expression of genes associated with cell junctions, cell cycle, oxidative stress and CC serpins. No differences were observed between the control and uPA group in cleavage rate or in blastocyst yield recorded on Days 7, 8 or 9 post-insemination. However, amiloride led to drastically reduced cleavage rate (28.5% vs 83.2%) and Day 9 embryo production (6.0% vs 21.0%) over the rates recorded for DMSO. These results indicate that the proteolytic activity of uPA is needed for successful oocyte maturation in bovine.
Fil: Roldan Olarte, Eugenia Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Maillo, Verónica. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Sánchez Calabuig, María Jesús. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Beltrán Breña, Paula. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Rizos, Dimitrios. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Gutiérrez Adan, Alfonso. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Materia
OOCYTE
UROKINASE
AMILORIDE
IN VITRO MATURATION
Nivel de accesibilidad
acceso embargado
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:11336/53719
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/53719
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturationRoldan Olarte, Eugenia MarielaMaillo, VerónicaSánchez Calabuig, María JesúsBeltrán Breña, PaulaRizos, DimitriosGutiérrez Adan, AlfonsoOOCYTEUROKINASEAMILORIDEIN VITRO MATURATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1This study examines the impacts of the urokinase-type plasminogen activator (uPA) on the in vitro maturation (IVM) of bovine oocytes. Cumulus–oocyte complexes in IVM medium were treated with uPA, amiloride (an uPA inhibitor), dimethyl sulfoxide (DMSO) or left untreated (control group). After 24 h of IVM, oocytes were recovered for testing or were in vitro fertilized and cultured to the blastocyst stage. The factors examined in all groups were: (i) oocyte nuclear maturation (Hoëscht staining); (ii) oocyte cytoplasmic maturation (cortical granules, CGs, distribution assessed by LCA-FITC); (iii) oocyte and cumulus cell (CC) gene expression (RT-qPCR); and (iv) embryo development (cleavage rate and blastocyst yield). Oocytes subjected to uPA treatment showed rates of nuclear maturation and CG distribution patterns similar to controls (P> 0.05), whereas lower rates of oocyte maturation were recorded in the amiloride group (P< 0.05). Both in oocytes and CC, treatment with uPA did not affect the transcription of genes related to apoptosis, cell junctions, cell cycle or serpin protease inhibitors. In contrast, amiloride altered the expression of genes associated with cell junctions, cell cycle, oxidative stress and CC serpins. No differences were observed between the control and uPA group in cleavage rate or in blastocyst yield recorded on Days 7, 8 or 9 post-insemination. However, amiloride led to drastically reduced cleavage rate (28.5% vs 83.2%) and Day 9 embryo production (6.0% vs 21.0%) over the rates recorded for DMSO. These results indicate that the proteolytic activity of uPA is needed for successful oocyte maturation in bovine.Fil: Roldan Olarte, Eugenia Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaFil: Maillo, Verónica. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaFil: Sánchez Calabuig, María Jesús. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaFil: Beltrán Breña, Paula. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaFil: Rizos, Dimitrios. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaFil: Gutiérrez Adan, Alfonso. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaBioScientifica2017-09info:eu-repo/date/embargoEnd/2018-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53719Roldan Olarte, Eugenia Mariela; Maillo, Verónica; Sánchez Calabuig, María Jesús; Beltrán Breña, Paula; Rizos, Dimitrios; et al.; Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation; BioScientifica; Reproduction; 154; 3; 9-2017; 331-3401470-1626CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.reproduction-online.org/content/154/3/331info:eu-repo/semantics/altIdentifier/doi/10.1530/REP-17-0204info:eu-repo/semantics/embargoedAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:52:09Zoai:ri.conicet.gov.ar:11336/53719instacron: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 09:52:09.697CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
title Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
spellingShingle Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
Roldan Olarte, Eugenia Mariela
OOCYTE
UROKINASE
AMILORIDE
IN VITRO MATURATION
title_short Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
title_full Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
title_fullStr Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
title_full_unstemmed Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
title_sort Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation
dc.creator.none.fl_str_mv Roldan Olarte, Eugenia Mariela
Maillo, Verónica
Sánchez Calabuig, María Jesús
Beltrán Breña, Paula
Rizos, Dimitrios
Gutiérrez Adan, Alfonso
author Roldan Olarte, Eugenia Mariela
author_facet Roldan Olarte, Eugenia Mariela
Maillo, Verónica
Sánchez Calabuig, María Jesús
Beltrán Breña, Paula
Rizos, Dimitrios
Gutiérrez Adan, Alfonso
author_role author
author2 Maillo, Verónica
Sánchez Calabuig, María Jesús
Beltrán Breña, Paula
Rizos, Dimitrios
Gutiérrez Adan, Alfonso
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv OOCYTE
UROKINASE
AMILORIDE
IN VITRO MATURATION
topic OOCYTE
UROKINASE
AMILORIDE
IN VITRO MATURATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv This study examines the impacts of the urokinase-type plasminogen activator (uPA) on the in vitro maturation (IVM) of bovine oocytes. Cumulus–oocyte complexes in IVM medium were treated with uPA, amiloride (an uPA inhibitor), dimethyl sulfoxide (DMSO) or left untreated (control group). After 24 h of IVM, oocytes were recovered for testing or were in vitro fertilized and cultured to the blastocyst stage. The factors examined in all groups were: (i) oocyte nuclear maturation (Hoëscht staining); (ii) oocyte cytoplasmic maturation (cortical granules, CGs, distribution assessed by LCA-FITC); (iii) oocyte and cumulus cell (CC) gene expression (RT-qPCR); and (iv) embryo development (cleavage rate and blastocyst yield). Oocytes subjected to uPA treatment showed rates of nuclear maturation and CG distribution patterns similar to controls (P> 0.05), whereas lower rates of oocyte maturation were recorded in the amiloride group (P< 0.05). Both in oocytes and CC, treatment with uPA did not affect the transcription of genes related to apoptosis, cell junctions, cell cycle or serpin protease inhibitors. In contrast, amiloride altered the expression of genes associated with cell junctions, cell cycle, oxidative stress and CC serpins. No differences were observed between the control and uPA group in cleavage rate or in blastocyst yield recorded on Days 7, 8 or 9 post-insemination. However, amiloride led to drastically reduced cleavage rate (28.5% vs 83.2%) and Day 9 embryo production (6.0% vs 21.0%) over the rates recorded for DMSO. These results indicate that the proteolytic activity of uPA is needed for successful oocyte maturation in bovine.
Fil: Roldan Olarte, Eugenia Mariela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Investigaciones Biológicas. Universidad Nacional de Tucumán. Instituto Superior de Investigaciones Biológicas; Argentina. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Maillo, Verónica. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Sánchez Calabuig, María Jesús. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Beltrán Breña, Paula. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Rizos, Dimitrios. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Fil: Gutiérrez Adan, Alfonso. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
description This study examines the impacts of the urokinase-type plasminogen activator (uPA) on the in vitro maturation (IVM) of bovine oocytes. Cumulus–oocyte complexes in IVM medium were treated with uPA, amiloride (an uPA inhibitor), dimethyl sulfoxide (DMSO) or left untreated (control group). After 24 h of IVM, oocytes were recovered for testing or were in vitro fertilized and cultured to the blastocyst stage. The factors examined in all groups were: (i) oocyte nuclear maturation (Hoëscht staining); (ii) oocyte cytoplasmic maturation (cortical granules, CGs, distribution assessed by LCA-FITC); (iii) oocyte and cumulus cell (CC) gene expression (RT-qPCR); and (iv) embryo development (cleavage rate and blastocyst yield). Oocytes subjected to uPA treatment showed rates of nuclear maturation and CG distribution patterns similar to controls (P> 0.05), whereas lower rates of oocyte maturation were recorded in the amiloride group (P< 0.05). Both in oocytes and CC, treatment with uPA did not affect the transcription of genes related to apoptosis, cell junctions, cell cycle or serpin protease inhibitors. In contrast, amiloride altered the expression of genes associated with cell junctions, cell cycle, oxidative stress and CC serpins. No differences were observed between the control and uPA group in cleavage rate or in blastocyst yield recorded on Days 7, 8 or 9 post-insemination. However, amiloride led to drastically reduced cleavage rate (28.5% vs 83.2%) and Day 9 embryo production (6.0% vs 21.0%) over the rates recorded for DMSO. These results indicate that the proteolytic activity of uPA is needed for successful oocyte maturation in bovine.
publishDate 2017
dc.date.none.fl_str_mv 2017-09
info:eu-repo/date/embargoEnd/2018-10-01
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/11336/53719
Roldan Olarte, Eugenia Mariela; Maillo, Verónica; Sánchez Calabuig, María Jesús; Beltrán Breña, Paula; Rizos, Dimitrios; et al.; Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation; BioScientifica; Reproduction; 154; 3; 9-2017; 331-340
1470-1626
CONICET Digital
CONICET
url http://hdl.handle.net/11336/53719
identifier_str_mv Roldan Olarte, Eugenia Mariela; Maillo, Verónica; Sánchez Calabuig, María Jesús; Beltrán Breña, Paula; Rizos, Dimitrios; et al.; Effect of urokinase type plasminogen activator on in vitro bovine oocyte maturation; BioScientifica; Reproduction; 154; 3; 9-2017; 331-340
1470-1626
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.reproduction-online.org/content/154/3/331
info:eu-repo/semantics/altIdentifier/doi/10.1530/REP-17-0204
dc.rights.none.fl_str_mv info:eu-repo/semantics/embargoedAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv embargoedAccess
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.publisher.none.fl_str_mv BioScientifica
publisher.none.fl_str_mv BioScientifica
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 13.13397

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