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

Canizo, Jésica; Rivolta, A. E. Ynsaurralde; Echegaray, Camila Vázquez; Suvá, M.; Alberio, V.; Aller, Juan F.; Guberman, Alejandra; Salamone, D.; Alberio, R. H.; Alberio, Ramiro

doi:10.1186/s12861-019-0193-9

Cited by 29 publications

(64 citation statements)

References 37 publications

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“…In contrast to PHD medium, N2B27 allows both complete hypoblast migration and epiblast development. Hypoblast proliferation, previously reported in the PHD system based on conventional histology and electron microscopy ( Brandao et al 2004 ), was confirmed by immunostaining for SOX17, an hypoblast marker ( Canizo et al 2019 ). However, irrespective of the substrate used, hypoblast cells did not cover the entire surface of the embryos developed in PHD medium, while all embryos developed in N2B27 medium showed complete hypoblast migration.…”

Section: Discussionsupporting

confidence: 63%

Embryonic disc formation following post-hatching bovine embryo development in vitro

et al. 2020

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Failures during conceptus elongation are a major cause of pregnancy losses in ungulates, exerting a relevant economic impact on farming. The developmental events occurring during this period are poorly understood, mainly because this process cannot be recapitulated in vitro. Previous studies have established an in vitro post-hatching development (PHD) system that supports bovine embryo development beyond the blastocyst stage, based on agarose gel tunnels and serum and glucose-enriched medium. Unfortunately, under this system embryonic disc formation is not achieved and embryos show notorious signs of apoptosis and necrosis. The objective of this study has been to develop an in vitro system able to support embryonic disc formation. We first compared post-hatching development inside agarose tunnels or free-floating over an agarose-coated dish in serum and glucose-enriched medium (PHD medium). Culture inside agarose tunnels shaped embryo morphology by physical constriction, but it restricted embryo growth and did not provide any significant advantage in terms of development of hypoblast and epiblast lineages. In contrast to PHD medium, a chemically defined and enriched medium (N2B27) supported complete hypoblast migration and epiblast survival in vitro, even in the absence of agarose coating. Cells expressing the pluripotency marker SOX2 were observed in ~56 % of the embryos and ~25 % developed embryonic disc-like structures formed by SOX2+ cells. In summary, here we provide a culture system that supports trophectoderm proliferation, hypoblast migration and epiblast survival after the blastocyst stage.

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Section: Discussionsupporting

confidence: 63%

Embryonic disc formation following post-hatching bovine embryo development in vitro

et al. 2020

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“…Later, during the second lineage segregation, ERK inhibition abrogates hypoblast formation in mouse and rabbit embryos ( Nichols et al., 2009 ; Piliszek et al., 2017 ). In contrast, in human ( Kuijk et al., 2012 ; Roode et al., 2012 ), pig ( Rodriguez et al., 2012 ), and cattle ( Canizo et al., 2019 ; Kuijk et al., 2012 ), ERK inhibition only partially blocks the hypoblast program, suggesting that additional signals regulate segregation of this lineage in non-murine embryos.…”

Section: Peri-implantation Development In Mammalian Embryos With Bilaminar Discsmentioning

confidence: 99%

“…Functional complementarity between human and other large mammalian embryos also find parallels in the signaling principles governing lineage decisions in the early embryo. During the first lineage decision segregating the TE and ICM, for example, inhibition of ERK signaling from the morula stage in pig and cattle promotes expansion of the TE in blastocysts (Canizo et al, 2019;Rodriguez et al, 2012). Although information on how human embryos respond to this treatment is lacking, experiments using naive human ESCs show that they readily differentiate into TE upon ERK inhibition alone (Guo et al, 2020).…”

Section: Reviewmentioning

confidence: 99%

Conserved features of non-primate bilaminar disc embryos and the germline

2021

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Post-implantation embryo development commences with a bilaminar disc in most mammals, including humans. Whereas access to early human embryos is limited and subject to greater ethical scrutiny, studies on non-primate embryos developing as bilaminar discs offer exceptional opportunities for advances in gastrulation, the germline, and the basis for evolutionary divergence applicable to human development. Here, we discuss the advantages of investigations in the pig embryo as an exemplar of development of a bilaminar disc embryo with relevance to early human development. Besides, the pig has the potential for the creation of humanized organs for xenotransplantation. Precise genetic engineering approaches, imaging, and single-cell analysis are cost effective and efficient, enabling research into some outstanding questions on human development and for developing authentic models of early human development with stem cells.

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“…On the contrary, expression of NANOG fails in OCT4-deficient bovine blastocysts, while the early presumptive hypoblast marker GATA6 is still present. Yet, it remains unclear if OCT4 has a role in the second lineage differentiation in bovine embryos, as GATA6 does not exclusively mark cells committed to the hypoblast, but also cells in the TE [9,10].…”

Section: Introductionmentioning

confidence: 99%

The second lineage differentiation of bovine embryos fails in the absence of OCT4/POU5F1

Simmet¹,

Kurome²,

Zakhartchenko³

et al. 2021

Preprint

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The mammalian blastocyst undergoes two lineage segregations, i.e., formation of the trophectoderm and subsequently differentiation of the hypoblast (HB) from the inner cell mass, leaving the epiblast (EPI) the remaining pluripotent lineage. To clarify expression patterns of markers specific for these lineages in bovine embryos, we analyzed day 7, 9 and 12 blastocysts completely derived ex vivo by staining for OCT4, NANOG, SOX2 (EPI) and GATA6, SOX17 (HB) and identified genes specific for these developmental stages in a global transcriptomics approach. To study the role of OCT4, we generated OCT4-deficient (OCT4 KO) embryos via somatic cell nuclear transfer or in vitro fertilization. OCT4 KO embryos reached the expanded blastocyst stage by day 8 but lost of NANOG and SOX17 expression, while SOX2 and GATA6 were unaffected. Blastocysts transferred to recipient cows from day 6 to 9 expanded, but the OCT4 KO phenotype was not rescued by the uterine environment. Exposure of OCT4 KO embryos to exogenous FGF4 or chimeric complementation with OCT4 intact embryos did not restore NANOG or SOX17 in OCT4-deficient cells. Our data show, that OCT4 is required cell-autonomously for the maintenance of pluripotency of the EPI and differentiation of the HB in bovine embryos.

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

Cited by 29 publications

References 37 publications

Embryonic disc formation following post-hatching bovine embryo development in vitro

Embryonic disc formation following post-hatching bovine embryo development in vitro

Conserved features of non-primate bilaminar disc embryos and the germline

The second lineage differentiation of bovine embryos fails in the absence of OCT4/POU5F1

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