A New Toolbox in Experimental Embryology—Alternative Model Organisms for Studying Preimplantation Development

Springer, Claudia; Wolf, Eckhard; Simmet, Kilian

doi:10.3390/jdb9020015

Cited by 4 publications

(10 citation statements)

References 205 publications

(228 reference statements)

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“…To investigate the regulation of differentiation and maintenance of pluripotency during mammalian preimplantation development, it is vital to examine models other than mouse, as species-specific differences exist. The bovine embryo is a very suitable alternative, as IVP procedures are highly developed and similarities to human embryo development have been reported (reviewed in [ 23 ]).…”

Section: Discussionmentioning

confidence: 99%

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Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

Springer

Zakhartchenko

Wolf

et al. 2021

Cells

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The role of the pluripotency factor NANOG during the second embryonic lineage differentiation has been studied extensively in mouse, although species-specific differences exist. To elucidate the role of NANOG in an alternative model organism, we knocked out NANOG in fibroblast cells and produced bovine NANOG-knockout (KO) embryos via somatic cell nuclear transfer (SCNT). At day 8, NANOG-KO blastocysts showed a decreased total cell number when compared to controls from SCNT (NT Ctrl). The pluripotency factors OCT4 and SOX2 as well as the hypoblast (HB) marker GATA6 were co-expressed in all cells of the inner cell mass (ICM) and, in contrast to mouse Nanog-KO, expression of the late HB marker SOX17 was still present. We blocked the MEK-pathway with a MEK 1/2 inhibitor, and control embryos showed an increase in NANOG positive cells, but SOX17 expressing HB precursor cells were still present. NANOG-KO together with MEK-inhibition was lethal before blastocyst stage, similarly to findings in mouse. Supplementation of exogenous FGF4 to NANOG-KO embryos did not change SOX17 expression in the ICM, unlike mouse Nanog-KO embryos, where missing SOX17 expression was completely rescued by FGF4. We conclude that NANOG mediated FGF/MEK signaling is not required for HB formation in the bovine embryo and that another—so far unknown—pathway regulates HB differentiation.

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Section: Discussionmentioning

confidence: 99%

“…Supplementing exogenous FGF4 from morula to blastocyst stage leads to ubiquitous GATA6 expression in mouse, bovine, pig and rabbit embryos (reviewed in [ 23 ]), revealing a non-cell-autonomous role for FGF4.…”

Section: Introductionmentioning

confidence: 99%

Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

Springer

Zakhartchenko

Wolf

et al. 2021

Cells

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“…The second lineage differentiation, when the PE/HB and EPI segregate, is regulated by EPI precursor cells expressing FGF4, which via the MEK‐pathway induces the differentiation of PE/HB precursor cells. Preimplantation embryos cultured with exogenous FGF4 develop an ICM entirely made up of PE/HB cells 4 6 .…”

Section: Introductionmentioning

confidence: 99%

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

et al. 2022

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The mammalian blastocyst undergoes two lineage segregations, that is, formation of the trophectoderm and subsequently differentiation of the hypoblast (HB) from the inner cell mass, leaving the epiblast (EPI) as the remaining pluripotent lineage. To clarify the expression patterns of markers specific for these lineages in bovine embryos, we analyzed day 7, 9, and 12 blastocysts completely produced in 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 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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“…Given the substantial differences in regulation of lineage differentiation and maintenance of pluripotency between mouse and other mammalian species, this progress harbors the prospect of a deeper understanding of preimplantation development, also in human. Because in vitro embryo production techniques are highly advanced in bovine, this species offers great opportunities as a model for preimplantation development [3][4][5].…”

Section: Introductionmentioning

confidence: 99%

“…The second lineage differentiation, when the PE/HB and EPI segregate, is regulated by EPI precursor cells expressing FGF4, which via the MEK-pathway induces the differentiation of PE/HB precursor cells. Preimplantation embryos cultured with exogenous FGF4 develop an ICM entirely made up of PE/HB cells [4]. The transcription factor OCT4/POU5F1 plays a pivotal role in mammalian embryo development, as it regulates both the maintenance of pluripotency as well as differentiation events [6].…”

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 New Toolbox in Experimental Embryology—Alternative Model Organisms for Studying Preimplantation Development

Cited by 4 publications

References 205 publications

Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

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

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