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The effect of dual inhibition of Ras–MEK–ERK and GSK3β pathways on development of in vitro cultured rabbit embryos

Published online by Cambridge University Press:  20 March 2020

Babett Bontovics ,
Pouneh Maraghechi ,
Bence Lázár ,
Mahek Anand ,
Kinga Németh ,
Renáta Fábián ,
Jaromír Vašíček ,
Alexander V. Makarevich ,
Elen Gócza Open the ORCID record for Elen Gócza [Opens in a new window]  and
Peter Chrenek
Babett Bontovics
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Pouneh Maraghechi
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Bence Lázár
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Mahek Anand
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Kinga Németh
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary Department of Laboratory Medicine, Semmelweis University, 1088Budapest, Szentkiralyi str. 46, Hungary
Renáta Fábián
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Jaromír Vašíček
Affiliation:
Research Institute for Animal Production Nitra, NPPC, Hlohovecka 2, 951 41Lužianky, Slovak Republic Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Hlinku 2, 949 76, Nitra, Slovak Republic
Alexander V. Makarevich
Affiliation:
Research Institute for Animal Production Nitra, NPPC, Hlohovecka 2, 951 41Lužianky, Slovak Republic
Elen Gócza*
Affiliation:
NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100Gödöllő, Szent-Györgyi A. str. 4., Hungary
Peter Chrenek
Affiliation:
Research Institute for Animal Production Nitra, NPPC, Hlohovecka 2, 951 41Lužianky, Slovak Republic Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Hlinku 2, 949 76, Nitra, Slovak Republic Faculty of Animal Breeding and Biology, UTP University of Science and Technology, Mazowiecka 28, 85-084Bydgoszcz, Poland
*
Author for correspondence: Elen Gócza, NARIC, Agricultural Biotechnology Institute, Animal Biotechnology Department, 2100 Gödöllő, Szent-Györgyi A. str. 4., Hungary Tel: +36 28 526 162. Fax: +36 28 526 151. E-mail: gocza.elen@abc.naik.hu
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Summary

Dual inhibition (2i) of Ras–MEK–ERK and GSK3β pathways enables the derivation of embryo stem cells (ESCs) from refractory mouse strains and, for permissive strains, allows ESC derivation with no external protein factor stimuli involvement. In addition, blocking of ERK signalling in 8-cell-stage mouse embryos leads to ablation of GATA4/6 expression in hypoblasts, suggesting fibroblast growth factor (FGF) dependence of hypoblast formation in the mouse. In human, bovine or porcine embryos, the hypoblast remains unaffected or displays slight-to-moderate reduction in cell number. In this study, we demonstrated that segregation of the hypoblast and the epiblast in rabbit embryos is FGF independent and 2i treatment elicits only a limited reinforcement in favour of OCT4-positive epiblast populations against the GATA4-/6-positive hypoblast population. It has been previously shown that TGFβ/Activin A inhibition overcomes the pervasive differentiation and inhomogeneity of rat iPSCs, rat ESCs and human iPSCs while prompting them to acquire naïve properties. However, TGFβ/Activin A inhibition, alone or together with Rho-associated, coiled-coil containing protein kinase (ROCK) inhibition, was not compatible with the viability of rabbit embryos according to the ultrastructural analysis of preimplantation rabbit embryos by electron microscopy. In rabbit models ovulation upon mating allows the precise timing of progression of the pregnancy. It produces several embryos of the desired stage in one pregnancy and a relatively short gestation period, making the rabbit embryo a suitable model to discover the cellular functions and mechanisms of maintenance of pluripotency in embryonic cells and the embryo-derived stem cells of other mammals.

Keywords

BlastocystEpiblastGATA6OCT4Small molecule inhibition
Type
Research Article
Information
Zygote , Volume 28 , Issue 3 , June 2020 , pp. 183 - 190
Copyright
© Cambridge University Press 2020

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Footnotes

*

These authors contributed equally to this work.

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