A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos

Autores
Canizo, Jésica Romina; Ynsaurralde Rivolta, Amanda Eugenia; Vazquez Echegaray, Camila; Suvá, Mariana; Alberio, Virgilia; Aller Atucha, Juan Florencio; Guberman, Alejandra; Salamone, Daniel Felipe; Alberio, Ricardo; Alberio, Ramiro
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: The segregation of the hypoblast and the emergence of the pluripotent epiblast mark the final stages of blastocyst formation in mammalian embryos. In bovine embryos the formation of the hypoblast has been partially studied, and evidence shows that MEK signalling plays a limited role in the segregation of this lineage. Here we explored the role of different signalling pathways during lineage segregation in the bovine embryo using immunofluorescence analysis of NANOG and SOX17 as readouts of epiblast and hypoblast, respectively. Results We show that SOX17 starts to be expressed in 16–32-cell stage embryos, whereas NANOG is first detected from 8-cell stage. SOX17 is first co-expressed with NANOG, but these markers become mutually exclusive by the late blastocyst stage. By assessing the expression kinetics of NANOG/SOX17 we show that inhibition of MEK signalling can eliminate SOX17 expression in bovine blastocysts, without altering NANOG expression. Modulation of WNT, PKC and LIF did not affect NANOG expression in the epiblast when used in combination with the ERK inhibitor. Conclusions This study shows that SOX17 can be used as a reliable early marker of hypoblast in the bovine, and based on its expression profile we show that the hypoblast segregates in day 7 blastocysts. Furthermore, SOX17 expression is abolished using 1 μM of PD0325901, without affecting the NANOG population in the epiblast. Modulation of WNT, PKC and LIF are not sufficient to support enhanced NANOG expression in the epiblast when combined with ERK inhibitor, indicating that additional signalling pathways should be examined to determine their potential roles in epiblast expansion.
EEA Balcarce
Fil: Canizo, Jesica Romina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil: Ynsaurralde Rivolta, Amada Eugenia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vazquez Echegaray, Camila. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Suvá, Mariana. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alberio, Virgilia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aller, Juan Florencio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.
Fil: Guberman, Alejandra S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Salamone, Daniel.F. Universidad de Buenos Aires. Facultad de Agronomía; Argentina.
Fil: Alberio, Ricardo H. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina.
Fil: Alberio, Ramiro. University of Nottingham. School of Biosciences; Reino Unido
Fuente
BMC Developmental Biology 19 : 13 (2019)
Materia
Ganado Bovino
Embriones Animales
Segregación
Linaje
Inmunofluorescencia
Cattle
Animal Embryos
Segregation
Lineage
Immunofluorescence
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/6009
Acceder
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oai_identifier_str oai:localhost:20.500.12123/6009
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryosCanizo, Jésica RominaYnsaurralde Rivolta, Amanda EugeniaVazquez Echegaray, CamilaSuvá, MarianaAlberio, VirgiliaAller Atucha, Juan FlorencioGuberman, AlejandraSalamone, Daniel FelipeAlberio, RicardoAlberio, RamiroGanado BovinoEmbriones AnimalesSegregaciónLinajeInmunofluorescenciaCattleAnimal EmbryosSegregationLineageImmunofluorescenceBackground: The segregation of the hypoblast and the emergence of the pluripotent epiblast mark the final stages of blastocyst formation in mammalian embryos. In bovine embryos the formation of the hypoblast has been partially studied, and evidence shows that MEK signalling plays a limited role in the segregation of this lineage. Here we explored the role of different signalling pathways during lineage segregation in the bovine embryo using immunofluorescence analysis of NANOG and SOX17 as readouts of epiblast and hypoblast, respectively. Results We show that SOX17 starts to be expressed in 16–32-cell stage embryos, whereas NANOG is first detected from 8-cell stage. SOX17 is first co-expressed with NANOG, but these markers become mutually exclusive by the late blastocyst stage. By assessing the expression kinetics of NANOG/SOX17 we show that inhibition of MEK signalling can eliminate SOX17 expression in bovine blastocysts, without altering NANOG expression. Modulation of WNT, PKC and LIF did not affect NANOG expression in the epiblast when used in combination with the ERK inhibitor. Conclusions This study shows that SOX17 can be used as a reliable early marker of hypoblast in the bovine, and based on its expression profile we show that the hypoblast segregates in day 7 blastocysts. Furthermore, SOX17 expression is abolished using 1 μM of PD0325901, without affecting the NANOG population in the epiblast. Modulation of WNT, PKC and LIF are not sufficient to support enhanced NANOG expression in the epiblast when combined with ERK inhibitor, indicating that additional signalling pathways should be examined to determine their potential roles in epiblast expansion.EEA BalcarceFil: Canizo, Jesica Romina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil: Ynsaurralde Rivolta, Amada Eugenia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Vazquez Echegaray, Camila. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Suvá, Mariana. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alberio, Virgilia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Aller, Juan Florencio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.Fil: Guberman, Alejandra S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Salamone, Daniel.F. Universidad de Buenos Aires. Facultad de Agronomía; Argentina.Fil: Alberio, Ricardo H. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina.Fil: Alberio, Ramiro. University of Nottingham. School of Biosciences; Reino UnidoBioMed Central2019-09-30T11:08:27Z2019-09-30T11:08:27Z2019-07-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/6009https://bmcdevbiol.biomedcentral.com/articles/10.1186/s12861-019-0193-91471-213Xhttps://doi.org/10.1186/s12861-019-0193-9BMC Developmental Biology 19 : 13 (2019)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-10-23T11:17:05Zoai:localhost:20.500.12123/6009instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-10-23 11:17:05.631INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
title A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
spellingShingle A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
Canizo, Jésica Romina
Ganado Bovino
Embriones Animales
Segregación
Linaje
Inmunofluorescencia
Cattle
Animal Embryos
Segregation
Lineage
Immunofluorescence
title_short A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
title_full A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
title_fullStr A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
title_full_unstemmed A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
title_sort A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
dc.creator.none.fl_str_mv Canizo, Jésica Romina
Ynsaurralde Rivolta, Amanda Eugenia
Vazquez Echegaray, Camila
Suvá, Mariana
Alberio, Virgilia
Aller Atucha, Juan Florencio
Guberman, Alejandra
Salamone, Daniel Felipe
Alberio, Ricardo
Alberio, Ramiro
author Canizo, Jésica Romina
author_facet Canizo, Jésica Romina
Ynsaurralde Rivolta, Amanda Eugenia
Vazquez Echegaray, Camila
Suvá, Mariana
Alberio, Virgilia
Aller Atucha, Juan Florencio
Guberman, Alejandra
Salamone, Daniel Felipe
Alberio, Ricardo
Alberio, Ramiro
author_role author
author2 Ynsaurralde Rivolta, Amanda Eugenia
Vazquez Echegaray, Camila
Suvá, Mariana
Alberio, Virgilia
Aller Atucha, Juan Florencio
Guberman, Alejandra
Salamone, Daniel Felipe
Alberio, Ricardo
Alberio, Ramiro
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Ganado Bovino
Embriones Animales
Segregación
Linaje
Inmunofluorescencia
Cattle
Animal Embryos
Segregation
Lineage
Immunofluorescence
topic Ganado Bovino
Embriones Animales
Segregación
Linaje
Inmunofluorescencia
Cattle
Animal Embryos
Segregation
Lineage
Immunofluorescence
dc.description.none.fl_txt_mv Background: The segregation of the hypoblast and the emergence of the pluripotent epiblast mark the final stages of blastocyst formation in mammalian embryos. In bovine embryos the formation of the hypoblast has been partially studied, and evidence shows that MEK signalling plays a limited role in the segregation of this lineage. Here we explored the role of different signalling pathways during lineage segregation in the bovine embryo using immunofluorescence analysis of NANOG and SOX17 as readouts of epiblast and hypoblast, respectively. Results We show that SOX17 starts to be expressed in 16–32-cell stage embryos, whereas NANOG is first detected from 8-cell stage. SOX17 is first co-expressed with NANOG, but these markers become mutually exclusive by the late blastocyst stage. By assessing the expression kinetics of NANOG/SOX17 we show that inhibition of MEK signalling can eliminate SOX17 expression in bovine blastocysts, without altering NANOG expression. Modulation of WNT, PKC and LIF did not affect NANOG expression in the epiblast when used in combination with the ERK inhibitor. Conclusions This study shows that SOX17 can be used as a reliable early marker of hypoblast in the bovine, and based on its expression profile we show that the hypoblast segregates in day 7 blastocysts. Furthermore, SOX17 expression is abolished using 1 μM of PD0325901, without affecting the NANOG population in the epiblast. Modulation of WNT, PKC and LIF are not sufficient to support enhanced NANOG expression in the epiblast when combined with ERK inhibitor, indicating that additional signalling pathways should be examined to determine their potential roles in epiblast expansion.
EEA Balcarce
Fil: Canizo, Jesica Romina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
Fil: Ynsaurralde Rivolta, Amada Eugenia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Vazquez Echegaray, Camila. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Suvá, Mariana. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alberio, Virgilia. Universidad de Buenos Aires. Facultad de Agronomía; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Aller, Juan Florencio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina.
Fil: Guberman, Alejandra S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Salamone, Daniel.F. Universidad de Buenos Aires. Facultad de Agronomía; Argentina.
Fil: Alberio, Ricardo H. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina.
Fil: Alberio, Ramiro. University of Nottingham. School of Biosciences; Reino Unido
description Background: The segregation of the hypoblast and the emergence of the pluripotent epiblast mark the final stages of blastocyst formation in mammalian embryos. In bovine embryos the formation of the hypoblast has been partially studied, and evidence shows that MEK signalling plays a limited role in the segregation of this lineage. Here we explored the role of different signalling pathways during lineage segregation in the bovine embryo using immunofluorescence analysis of NANOG and SOX17 as readouts of epiblast and hypoblast, respectively. Results We show that SOX17 starts to be expressed in 16–32-cell stage embryos, whereas NANOG is first detected from 8-cell stage. SOX17 is first co-expressed with NANOG, but these markers become mutually exclusive by the late blastocyst stage. By assessing the expression kinetics of NANOG/SOX17 we show that inhibition of MEK signalling can eliminate SOX17 expression in bovine blastocysts, without altering NANOG expression. Modulation of WNT, PKC and LIF did not affect NANOG expression in the epiblast when used in combination with the ERK inhibitor. Conclusions This study shows that SOX17 can be used as a reliable early marker of hypoblast in the bovine, and based on its expression profile we show that the hypoblast segregates in day 7 blastocysts. Furthermore, SOX17 expression is abolished using 1 μM of PD0325901, without affecting the NANOG population in the epiblast. Modulation of WNT, PKC and LIF are not sufficient to support enhanced NANOG expression in the epiblast when combined with ERK inhibitor, indicating that additional signalling pathways should be examined to determine their potential roles in epiblast expansion.
publishDate 2019
dc.date.none.fl_str_mv 2019-09-30T11:08:27Z
2019-09-30T11:08:27Z
2019-07-04
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/20.500.12123/6009
https://bmcdevbiol.biomedcentral.com/articles/10.1186/s12861-019-0193-9
1471-213X
https://doi.org/10.1186/s12861-019-0193-9
url http://hdl.handle.net/20.500.12123/6009
https://bmcdevbiol.biomedcentral.com/articles/10.1186/s12861-019-0193-9
https://doi.org/10.1186/s12861-019-0193-9
identifier_str_mv 1471-213X
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv BMC Developmental Biology 19 : 13 (2019)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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