A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos
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 t...
| Autores principales: | , , , , , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
BioMed Central
2019
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| Materias: | |
| Acceso en línea: | 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 |
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| author | 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_browse | Alberio, Ramiro Alberio, Ricardo Alberio, Virgilia Aller Atucha, Juan Florencio Canizo, Jésica Romina Guberman, Alejandra Salamone, Daniel Felipe Suvá, Mariana Vazquez Echegaray, Camila Ynsaurralde Rivolta, Amanda Eugenia |
| 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_sort | Canizo, Jésica Romina |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA6009 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | BioMed Central |
| publisherStr | BioMed Central |
| record_format | dspace |
| spelling | INTA60092024-06-07T11:34:13Z A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos 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 Ganado Bovino Embriones Animales Segregación Linaje Inmunofluorescencia Cattle Animal Embryos Segregation Lineage Immunofluorescence 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 2019-09-30T11:08:27Z 2019-09-30T11:08:27Z 2019-07-04 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf BioMed Central BMC Developmental Biology 19 : 13 (2019) |
| spellingShingle | Ganado Bovino Embriones Animales Segregación Linaje Inmunofluorescencia Cattle Animal Embryos Segregation Lineage Immunofluorescence 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 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 |
| 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_short | A dose-dependent response to MEK inhibition determines hypoblast fate in bovine embryos |
| title_sort | dose dependent response to mek inhibition determines hypoblast fate in bovine embryos |
| topic | Ganado Bovino Embriones Animales Segregación Linaje Inmunofluorescencia Cattle Animal Embryos Segregation Lineage Immunofluorescence |
| 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 |
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