Estudio comparativo de la diferenciación de los primeros linajes celulares en embriones de Ovis orientalis aries y Mus musculus

Flores Borobia, Inés (2021). Estudio comparativo de la diferenciación de los primeros linajes celulares en embriones de Ovis orientalis aries y Mus musculus. Proyecto Fin de Carrera / Trabajo Fin de Grado, E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM), Madrid.

Description

Title: Estudio comparativo de la diferenciación de los primeros linajes celulares en embriones de Ovis orientalis aries y Mus musculus
Author/s:
  • Flores Borobia, Inés
Contributor/s:
  • García Rebollar, Pilar
  • Ramos Ibeas, Priscila
Item Type: Final Project
Degree: Grado en Biotecnología
Date: June 2021
Subjects:
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Producción Agraria
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Early embryonic losses represent a major factor affecting reproductive efficiency of farm animals and humans. In vitro produced embryos show high mortality rates during preimplantation development, when complex cell differentiation processes happen. The critical differences in embryo development between mice, the most studied mammalian model, and other mammals, have precluded the identification of the mechanisms governing early lineages differentiation in farm animals. Better understanding of these processes and identification of suitable lineage markers, as well as evaluation of development kinetics are necessary to assess embryo quality. The aim of this study was to characterize and compare development kinetics and the expression of early cell lineage markers in mouse and sheep preimplantation embryos. Sheep embryos were produced and cultivated in vitro until post blastocyst stage and in vivo derived mouse zygotes were recovered. Ovine embryos were sequentially fixed at days (D) 1.5, 3, 4, 5, 6, 7, 8, 10 and 12, and murine embryos at D1.5, 2, 2.5, 3.5 and 4.5. Embryos were analysed by immunostaining for SOX2 (epiblast); SOX17 or GATA6 (hypoblast) and CDX2 or GATA3 (trophoblast), and counterstained with DAPI. Z-stack imaging was performed and cells expressing each marker and total cells were counted. To compare markers expression between species, embryos were grouped according to total cell number in: 2c, 3-4c, 5-8c, 9-16c, 17-32c, early and late blastocysts. Sheep embryos showed asynchronous development, in contrast to mouse embryos. SOX2 expression started at 3-4c in sheep and was observed in some 2c mouse embryos, later disappearing until the 9-16c stage. In both species, SOX2 was restricted to the epiblast from the blastocyst stage onwards. SOX17 was detected from 3-4c stage in sheep and from 17-32c in mouse and was restricted to hypoblast cells from the blastocyst stage in both species. However, GATA6 expression started at 5-8c in both species but it was specific to hypoblast cells only in mouse blastocysts, while in sheep it was ubiquitously expressed until D10. Finally, trophectoderm markers were detected earlier in mouse than in sheep embryos and the % of GATA3+ and CDX2+/total cells was significantly higher in mouse than in sheep at all stages, suggesting that trophoblast specification occurs later in ungulates than in mice. In conclusion, we provide a development kinetics analysis and an expression timeline of well-known lineage markers in mouse and sheep embryos, remarking significant differences between species. While all analysed markers can be used to assess proper lineage commitment and embryo quality in mice, SOX2, SOX17, CDX2 and GATA3, but not GATA6, are lineage-specific in sheep blastocysts.

More information

Item ID: 69286
DC Identifier: https://oa.upm.es/69286/
OAI Identifier: oai:oa.upm.es:69286
Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 15 Dec 2021 12:44
Last Modified: 15 Feb 2022 23:30
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