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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Aggregation recovers developmental plasticity in mouse polyploid embryos

Hiroyuki Imai A B , Wataru Fujii C , Ken Takeshi Kusakabe A , Yasuo Kiso A and Kiyoshi Kano https://orcid.org/0000-0002-8967-9536 A B D
+ Author Affiliations
- Author Affiliations

A Laboratory of Veterinary Anatomy, Joint-Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi, 7538515, Japan.

B Laboratory of Developmental Biology, Joint-Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi, 7538515, Japan.

C Laboratory of Applied Genetics, Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8517, Japan.

D Corresponding author. Email: kanokiyo@yamaguchi-u.ac.jp

Reproduction, Fertility and Development 31(2) 404-411 https://doi.org/10.1071/RD18093
Submitted: 9 March 2018  Accepted: 21 July 2018   Published: 13 September 2018

Abstract

Tetraploid embryos normally develop into blastocysts and embryonic stem cells can be established from tetraploid blastocysts in mice. Thus, polyploidisation does not seem to be so harmful during preimplantation development. However, the mechanisms by which early mammalian development accepts polyploidisation are poorly understood. In this study, we aimed to elucidate the effect of polyploidisation on early mammalian development and to further comprehend its tolerance using hyperpolyploid embryos produced by repetitive whole genome duplication. We successfully established several types of polyploid embryos (tetraploid, octaploid and hexadecaploid) and studied their developmental potential in vitro. We demonstrated that all types of these polyploid embryos maintained the ability to develop to the blastocyst stage, which implies that mammalian cells might have basic cellular functions in implanted embryos, despite polyploidisation. However, the inner cell mass was absent in hexadecaploid blastocysts. To complement the total number of cells in blastocysts, a fused hexadecaploid embryo was produced by aggregating several hexadecaploid embryos. The results indicated that the fused hexadecaploid embryo finally recovered pluripotent cells in the blastocyst. Thus, our findings suggest that early mammalian embryos may have the tolerance and higher plasticity to adapt to hyperpolyploidisation for blastocyst formation, despite intense alteration of the genome volume.

Additional keywords : aggregated embryo, blastocyst, inner cell mass, polyploidy.


References

Berg, H. (1982). Biological implications of electric field effects: Part V. Fusion of blastomeres and blastocysts of mouse embryos. Bioelectrochem. Bioenerg. 9, 223–228.
Biological implications of electric field effects: Part V. Fusion of blastomeres and blastocysts of mouse embryos.Crossref | GoogleScholarGoogle Scholar |

Brodsky, V. Y., and Uryvaeva, I. V. (1985) ‘Genome Multiplication in Growth and Development: Biology of Polyploid and Polytene Cells’. (Cambridge University Press: London.)

Buemo, C. P., Gambini, A., Moro, L. N., Hiriart, M. I., Fernandez-Martin, R., Collas, P., and Salamone, D. F. (2016). Embryo aggregation in pig improves cloning efficiency and embryo quality. PLoS One 11, .
Embryo aggregation in pig improves cloning efficiency and embryo quality.Crossref | GoogleScholarGoogle Scholar |

Cleveland, B. M., and Weber, G. M. (2014). Ploidy effects on genes regulating growth mechanisms during fasting and refeeding in juvenile rainbow trout (Oncorhynchus mykiss). Mol. Cell. Endocrinol. 382, 139–149.
Ploidy effects on genes regulating growth mechanisms during fasting and refeeding in juvenile rainbow trout (Oncorhynchus mykiss).Crossref | GoogleScholarGoogle Scholar |

Dietrich, J. E., and Hiiragi, T. (2007). Stochastic patterning in the mouse pre-implantation embryo. Development 134, 4219–4231.
Stochastic patterning in the mouse pre-implantation embryo.Crossref | GoogleScholarGoogle Scholar |

Dyban, A., and Baranov, V. (1987) ‘Cytogenetics of Mammalian Embryonic Development’. (Oxford University Press: New York.)

Eakin, G. S., and Behringer, R. R. (2003). Tetraploid development in the mouse. Dev. Dyn. 228, 751–766.
Tetraploid development in the mouse.Crossref | GoogleScholarGoogle Scholar |

Evans, M. J., and Kaufman, M. H. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–156.
Establishment in culture of pluripotential cells from mouse embryos.Crossref | GoogleScholarGoogle Scholar |

Hayashi, K., Ohta, H., Kurimoto, K., Aramaki, S., and Saitou, M. (2011). Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells. Cell 146, 519–532.
Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells.Crossref | GoogleScholarGoogle Scholar |

Hayashi, K., Ogushi, S., Kurimoto, K., Shimamoto, S., Ohta, H., and Saitou, M. (2012). Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice. Science 338, 971–975.
Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice.Crossref | GoogleScholarGoogle Scholar |

Imai, H., Kano, K., Fujii, W., Takasawa, K., Wakitani, S., Hiyama, M., Nishino, K., Kusakabe, K. T., and Kiso, Y. (2015). Tetraploid embryonic stem cells maintain pluripotency and differentiation potency into three germ layers. PLoS One 10, .
Tetraploid embryonic stem cells maintain pluripotency and differentiation potency into three germ layers.Crossref | GoogleScholarGoogle Scholar |

Imai, H., Fujii, W., Kusakabe, K. T., Kiso, Y., and Kano, K. (2016). Effects of whole genome duplication on cell size and gene expression in mouse embryonic stem cells. J. Reprod. Dev. 62, 571–576.
Effects of whole genome duplication on cell size and gene expression in mouse embryonic stem cells.Crossref | GoogleScholarGoogle Scholar |

Ishiguro, N., Kano, K., Yamamoto, Y., and Taniguchi, K. (2005). Tetraploid cells of enhanced green fluorescent protein transgenic mice in tetraploid/diploid-chimeric embryos. J. Reprod. Dev. 51, 567–572.
Tetraploid cells of enhanced green fluorescent protein transgenic mice in tetraploid/diploid-chimeric embryos.Crossref | GoogleScholarGoogle Scholar |

James, R. M., Kaufman, M. H., Webb, S., and West, J. D. (1992). Electrofusion of mouse embryos results in uniform tetraploidy and not tetraploid/diploid mosaicism. Genet. Res. 60, 185–194.
Electrofusion of mouse embryos results in uniform tetraploidy and not tetraploid/diploid mosaicism.Crossref | GoogleScholarGoogle Scholar |

Kasahara, M. (2007). The 2R hypothesis: an update. Curr. Opin. Immunol. 19, 547–552.
The 2R hypothesis: an update.Crossref | GoogleScholarGoogle Scholar |

Kaufman, M. H. (1991). Histochemical identification of primordial germ cells and differentiation of the gonads in homozygous tetraploid mouse embryos. J. Anat. 179, 169–181.

Kaufman, M. H. (1992). Postcranial morphological features of homozygous tetraploid mouse embryos. J. Anat. 180, 521–534.

Kaufman, M. H., and Webb, S. (1990). Postimplantation development of tetraploid mouse embryos produced by electrofusion. Development 110, 1121–1132.

Kawaguchi, J., Kano, K., and Naito, K. (2009). Expression profiling of tetraploid mouse embryos in the developmental stages using a cDNA microarray analysis. J. Reprod. Dev. 55, 670–675.
Expression profiling of tetraploid mouse embryos in the developmental stages using a cDNA microarray analysis.Crossref | GoogleScholarGoogle Scholar |

Koizumi, N., and Fukuta, K. (1995). Preimplantation development of tetraploid mouse embryo produced by cytochalasin B. Exp. Anim. 44, 105–109.
Preimplantation development of tetraploid mouse embryo produced by cytochalasin B.Crossref | GoogleScholarGoogle Scholar |

Kubiak, J. Z., and Tarkowski, A. K. (1985). Electrofusion of mouse blastomeres. Exp. Cell Res. 157, 561–566.
Electrofusion of mouse blastomeres.Crossref | GoogleScholarGoogle Scholar |

Kurischko, A., and Berg, H. (1986). Electrofusion of rat and mouse blastomeres. Bioelectrochem. Bioenerg. 15, 513–519.
Electrofusion of rat and mouse blastomeres.Crossref | GoogleScholarGoogle Scholar |

Lin, M., Thorne, M. H., Martin, I. C., Sheldon, B. L., and Jones, R. C. (1995). Electron microscopy of the seminiferous epithelium in the triploid (ZZZ and ZZW) fowl, Gallus domesticus. J. Anat. 186, 563–576.

McFeely, R. A. (1969) Aneuploidy, polyploidy, and structural rearrangement of chromosomes in mammals other than man. In ‘Comparative Mammalian Cytogenetics’. (Ed. K. Benirschke.) pp. 434–444. (Springer-Verlag: Hanover, NH, USA.)

Ohno, S. (1970) ‘Evolution by Gene Duplication’. (Springer-Verlag: Berlin.)

Panopoulou, G., and Poustka, A. J. (2005). Timing and mechanism of ancient vertebrate genome duplications – the adventure of a hypothesis. Trends Genet. 21, 559–567.
Timing and mechanism of ancient vertebrate genome duplications – the adventure of a hypothesis.Crossref | GoogleScholarGoogle Scholar |

Pfeiffer, M. J., Esteves, T. C., Balbach, S. T., Arauzo-Bravo, M. J., Stehling, M., Jauch, A., Houghton, F. D., Schwarzer, C., and Boiani, M. (2013). Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells. Stem Cells 31, 2343–2353.
Reprogramming of two somatic nuclei in the same ooplasm leads to pluripotent embryonic stem cells.Crossref | GoogleScholarGoogle Scholar |

Piferrer, F., Beaumont, A., Falguière, J.-C., Flajšhans, M., Haffray, P., and Colombo, L. C. (2009). Polyploid fish and shellfish: production, biology and applications to aquaculture for performance improvement and genetic containment. Aquaculture 293, 125–156.
Polyploid fish and shellfish: production, biology and applications to aquaculture for performance improvement and genetic containment.Crossref | GoogleScholarGoogle Scholar |

Pralong, D., Lim, M. L., Vassiliev, I., Mrozik, K., Wijesundara, N., Rathjen, P., and Verma, P. J. (2005). Tetraploid embryonic stem cells contribute to the inner cell mass of mouse blastocysts. Cloning Stem Cells 7, 272–278.
Tetraploid embryonic stem cells contribute to the inner cell mass of mouse blastocysts.Crossref | GoogleScholarGoogle Scholar |

Snow, M. H. (1973). Tetraploid mouse embryos produced by cytochalasin B during cleavage. Nature 244, 513–515.
Tetraploid mouse embryos produced by cytochalasin B during cleavage.Crossref | GoogleScholarGoogle Scholar |

Snow, M. H. (1975). Embryonic development of tetraploid mice during the second half of gestation. J. Embryol. Exp. Morphol. 34, 707–721.

Tarkowski, A. K. (1959). Experiments on the development of isolated blastomeres of mouse eggs. Nature 184, 1286–1287.
Experiments on the development of isolated blastomeres of mouse eggs.Crossref | GoogleScholarGoogle Scholar |

Tarkowski, A. K., and Wroblewska, J. (1967). Development of blastomeres of mouse eggs isolated at the 4- and 8-cell stage. J. Embryol. Exp. Morphol. 18, 155–180.

Tarkowski, A. K., Witkowska, A., and Opas, J. (1977). Development of cytochalasin in B-induced tetraploid and diploid/tetraploid mosaic mouse embryos. J. Embryol. Exp. Morphol. 41, 47–64.

Wen, D., Saiz, N., Rosenwaks, Z., Hadjantonakis, A. K., and Rafii, S. (2014). Completely ES cell-derived mice produced by tetraploid complementation using inner cell mass (ICM) deficient blastocysts. PLoS One 9, .
Completely ES cell-derived mice produced by tetraploid complementation using inner cell mass (ICM) deficient blastocysts.Crossref | GoogleScholarGoogle Scholar |

Wennekamp, S., Mesecke, S., Nedelec, F., and Hiiragi, T. (2013). A self-organization framework for symmetry breaking in the mammalian embryo. Nat. Rev. Mol. Cell Biol. 14, 452–459.
A self-organization framework for symmetry breaking in the mammalian embryo.Crossref | GoogleScholarGoogle Scholar |

Wu, B. J., Zhao, L. X., Zhu, C. C., Chen, Y. L., Wei, M. Y., Bao, S. Q., Sun, S. C., and Li, X. H. (2017). Altered apoptosis/autophagy and epigenetic modifications cause the impaired postimplantation octaploid embryonic development in mice. Cell Cycle 16, 82–90.
Altered apoptosis/autophagy and epigenetic modifications cause the impaired postimplantation octaploid embryonic development in mice.Crossref | GoogleScholarGoogle Scholar |