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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Mitochondrial DNA replication is initiated at blastocyst formation in equine embryos

W. Karin Hendriks https://orcid.org/0000-0002-0275-9278 A G , Silvia Colleoni B , Cesare Galli B C , Damien B. B. P. Paris https://orcid.org/0000-0001-7329-8985 D , Ben Colenbrander A and Tom A. E. Stout A E F H
+ Author Affiliations
- Author Affiliations

A Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM Utrecht, Netherlands.

B Avantea, Laboratorio di Technologie della Riproduzione, Via Porcellasco 7f, 26100 Cremona, Italy.

C Fondazione Avantea, Via Porcellasco 7f, 26100 Cremona, Italy.

D Discipline of Biomedical Science, College of Public Health, Medical and Veterinary Sciences, James Cook University, Solander Drive, Townsville, Qld 4811, Australia.

E Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, Netherlands.

F Section of Reproduction, Faculty of Veterinary Science, University of Pretoria, Private Bag XO4, Onderstepoort 0110, South Africa.

G Present address: Hendriks EQ Repro Consultancy, Zutphensestraatweg 32, 6955 AH Ellecom, Netherlands.

H Corresponding author. Email: t.a.e.stout@uu.nl

Reproduction, Fertility and Development 31(3) 570-578 https://doi.org/10.1071/RD17387
Submitted: 24 September 2017  Accepted: 13 September 2018   Published: 14 November 2018

Abstract

Intracytoplasmic sperm injection is the technique of choice for equine IVF and, in a research setting, 18–36% of injected oocytes develop to blastocysts. However, blastocyst development in clinical programs is lower, presumably due to a combination of variable oocyte quality (e.g. from old mares), suboptimal culture conditions and marginal fertility of some stallions. Furthermore, mitochondrial constitution appears to be critical to developmental competence, and both maternal aging and in vitro embryo production (IVEP) negatively affect mitochondrial number and function in murine and bovine embryos. The present study examined the onset of mitochondrial (mt) DNA replication in equine embryos and investigated whether IVEP affects the timing of this important event, or the expression of genes required for mtDNA replication (i.e. mitochondrial transcription factor (TFAM), mtDNA polymerase γ subunit B (mtPOLB) and single-stranded DNA binding protein (SSB)). We also investigated whether developmental arrest was associated with low mtDNA copy number. mtDNA copy number increased (P < 0.01) between the early and expanded blastocyst stages both in vivo and in vitro, whereas the mtDNA : total DNA ratio was higher in in vitro-produced embryos (P = 0.041). Mitochondrial replication was preceded by an increase in TFAM but, unexpectedly, not mtPOLB or SSB expression. There was no association between embryonic arrest and lower mtDNA copy numbers.

Additional keywords: embryo development, gene expression, horse, IVF, oocyte.


References

Badr, H., Bongioni, G., Abdoon, A. S., Kandil, O., and Puglisi, R. (2007). Gene expression in the in vitro-produced preimplantation bovine embryos. Zygote 15, 355–367.
Gene expression in the in vitro-produced preimplantation bovine embryos.Crossref | GoogleScholarGoogle Scholar |

Barritt, J., Willadsen, S., Brenner, C., and Cohen, J. (2001). Cytoplasmic transfer in assisted reproduction. Hum. Reprod. Update 7, 428–435.
Cytoplasmic transfer in assisted reproduction.Crossref | GoogleScholarGoogle Scholar |

Bentov, Y., Yavorska, T., Esfandiari, N., Jurisicova, A., and Casper, R. F. (2011). The contribution of mitochondrial function to reproductive aging. J. Assist. Reprod. Genet. 28, 773–783.
The contribution of mitochondrial function to reproductive aging.Crossref | GoogleScholarGoogle Scholar |

Bowles, E. J., Lee, J. H., Alberio, R., Lloyd, R. E., Stekel, D., Campbell, K. H., and St John, J. C. (2007). Contrasting effects of in vitro fertilization and nuclear transfer on the expression of mtDNA replication factors. Genetics 176, 1511–1526.
Contrasting effects of in vitro fertilization and nuclear transfer on the expression of mtDNA replication factors.Crossref | GoogleScholarGoogle Scholar |

Cagnone, G. L., Tsai, T. S., Makanji, Y., Matthews, P., Gould, J., Bonkowski, M. S., Elgass, K. D., Wong, A. S., Wu, L. E., McKenzie, M., Sinclair, D. A., and St John, J. C. (2016). Restoration of normal embryogenesis by mitochondrial supplementation in pig oocytes exhibiting mitochondrial DNA deficiency. Sci. Rep. 6, 23229.
Restoration of normal embryogenesis by mitochondrial supplementation in pig oocytes exhibiting mitochondrial DNA deficiency.Crossref | GoogleScholarGoogle Scholar |

Carnevale, E. M. (2008). The mare model for follicular maturation and reproductive aging in the woman. Theriogenology 69, 23–30.
The mare model for follicular maturation and reproductive aging in the woman.Crossref | GoogleScholarGoogle Scholar |

Chiaratti, M. R., Garcia, B. M., Carvalho, K. F., Machado, T. S., da Ribeiro, F. K. S., and Macabelli, C. H. (2018). The role of mitochondria in the female germline: implications to fertility and inheritance of mitochondrial diseases. Cell Biol. Int. 42, 711–724.
The role of mitochondria in the female germline: implications to fertility and inheritance of mitochondrial diseases.Crossref | GoogleScholarGoogle Scholar |

El Shourbagy, S. H., Spikings, E. C., Freitas, M., and St John, J. C. (2006). Mitochondria directly influence fertilisation outcome in the pig. Reproduction 131, 233–245.
Mitochondria directly influence fertilisation outcome in the pig.Crossref | GoogleScholarGoogle Scholar |

Facucho-Oliveira, J. M., Alderson, J., Spikings, E. C., Egginton, S., and St John, J. C. (2007). Mitochondrial DNA replication during differentiation of murine embryonic stem cells. J. Cell Sci. 120, 4025–4034.
Mitochondrial DNA replication during differentiation of murine embryonic stem cells.Crossref | GoogleScholarGoogle Scholar |

Fragouli, E., Spath, K., Alfarawati, S., Kaper, F., Craig, A., Michel, C. E., Kokocinski, F., Cohen, J., Munne, S., and Wells, D. (2015). Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genet. 11, e1005241.
Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential.Crossref | GoogleScholarGoogle Scholar |

Galli, C., Crotti, G., Notari, C., Turini, P., Duchi, R., and Lazzari, G. (2001). Embryo production by ovum pick up from live donors. Theriogenology 55, 1341–1357.
Embryo production by ovum pick up from live donors.Crossref | GoogleScholarGoogle Scholar |

Galli, C., Colleoni, S., Duchi, R., Lagutina, I., and Lazzari, G. (2007). Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer. Anim. Reprod. Sci. 98, 39–55.
Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.Crossref | GoogleScholarGoogle Scholar |

Giritharan, G., Talbi, S., Donjacour, A., Di Sebastiano, F., Dobson, A. T., and Rinaudo, P. F. (2007). Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos. Reproduction 134, 63–72.
Effect of in vitro fertilization on gene expression and development of mouse preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |

Giritharan, G., Li, M. W., Di Sebastiano, F., Esteban, F. J., Horcajadas, J. A., Lloyd, K. C., Donjacour, A., Maltepe, E., and Rinaudo, P. F. (2010). Effect of ICSI on gene expression and development of mouse preimplantation embryos. Hum. Reprod. 25, 3012–3024.
Effect of ICSI on gene expression and development of mouse preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |

Grøndahl, C., and Hyttel, P. (1996). Nucleologenesis and ribonucleic acid synthesis in preimplantation equine embryos. Biol. Reprod. 55, 769–774.
Nucleologenesis and ribonucleic acid synthesis in preimplantation equine embryos.Crossref | GoogleScholarGoogle Scholar |

Hashimoto, S., Morimoto, N., Yamanaka, M., Matsumoto, H., Yamochi, T., Goto, H., Inoue, M., Nakaoka, Y., Shibahara, H., and Morimoto, Y. (2017). Quantitative and qualitative changes of mitochondria in human preimplantation embryos. J. Assist. Reprod. Genet. 34, 573–580.
Quantitative and qualitative changes of mitochondria in human preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |

Hendriks, W. K., Colleoni, S., Galli, C., Paris, D. B. B. P., Colenbrander, B., Roelen, B. A. J., and Stout, T. A. E. (2015). Maternal age and in vitro culture affect mitochondrial number and function in equine oocytes and embryos. Reprod. Fertil. Dev. 27, 957–968.
Maternal age and in vitro culture affect mitochondrial number and function in equine oocytes and embryos.Crossref | GoogleScholarGoogle Scholar |

Hinrichs, K. (2013). Assisted reproduction techniques in the horse. Reprod. Fertil. Dev. 25, 80–93.
Assisted reproduction techniques in the horse.Crossref | GoogleScholarGoogle Scholar |

Hinrichs, K., Schmidt, A. L., Friedman, P. P., Selgrath, J. P., and Martin, M. G. (1993). In vitro maturation of horse oocytes: characterization of chromatin configuration using fluorescence microscopy. Biol. Reprod. 48, 363–370.
In vitro maturation of horse oocytes: characterization of chromatin configuration using fluorescence microscopy.Crossref | GoogleScholarGoogle Scholar |

Kameyama, Y., Filion, F., Yoo, J. G., and Smith, L. C. (2007). Characterization of mitochondrial replication and transcription control during rat early development in vivo and in vitro. Reproduction 133, 423–432.
Characterization of mitochondrial replication and transcription control during rat early development in vivo and in vitro.Crossref | GoogleScholarGoogle Scholar |

Klein, J. P., and Moeschberger, M. L. (2003). ‘Survival Analysis. Techniques for Censored and Truncated Data.’ 2nd edn. (Springer-Verlag: New York.)

May-Panloup, P., Chretien, M. F., Malthiery, Y., and Reynier, P. (2007). Mitochondrial DNA in the oocyte and the developing embryo. Curr. Top. Dev. Biol. 77, 51–83.
Mitochondrial DNA in the oocyte and the developing embryo.Crossref | GoogleScholarGoogle Scholar |

McConnell, J. M., and Petrie, L. (2004). Mitochondrial DNA turnover occurs during preimplantation development and can be modulated by environmental factors. Reprod. Biomed. Online 9, 418–424.
Mitochondrial DNA turnover occurs during preimplantation development and can be modulated by environmental factors.Crossref | GoogleScholarGoogle Scholar |

Paris, D. B., Kuijk, E. W., Roelen, B. A., and Stout, T. A. (2011). Establishing reference genes for use in real-time quantitative PCR analysis of early equine embryos. Reprod. Fertil. Dev. 23, 353–363.
Establishing reference genes for use in real-time quantitative PCR analysis of early equine embryos.Crossref | GoogleScholarGoogle Scholar |

Pikó, L., and Taylor, K. D. (1987). Amounts of mitochondrial DNA and abundance of some mitochondrial gene transcripts in early mouse embryos. Dev. Biol. 123, 364–374.
Amounts of mitochondrial DNA and abundance of some mitochondrial gene transcripts in early mouse embryos.Crossref | GoogleScholarGoogle Scholar |

Rambags, B. P., Krijtenburg, P. J., van Drie, H. F., Lazzari, G., Galli, C., Pearson, P. L., Colenbrander, B., and Stout, T. A. E. (2005). Numerical chromosomal abnormalities in equine embryos produced in vivo and in vitro. Mol. Reprod. Dev. 72, 77–87.
Numerical chromosomal abnormalities in equine embryos produced in vivo and in vitro.Crossref | GoogleScholarGoogle Scholar |

Rambags, B. P., van Boxtel, D. C., Tharasanit, T., Lenstra, J. A., Colenbrander, B., and Stout, T. A. (2014). Advancing maternal age predisposes to mitochondrial damage and loss during maturation of equine oocytes in vitro. Theriogenology 81, 959–965.
Advancing maternal age predisposes to mitochondrial damage and loss during maturation of equine oocytes in vitro.Crossref | GoogleScholarGoogle Scholar |

Reynier, P., May-Panloup, P., Chretien, M. F., Morgan, C. J., Jean, M., Savagner, F., Barriere, P., and Malthiery, Y. (2001). Mitochondrial DNA content affects the fertilizability of human oocytes. Mol. Hum. Reprod. 7, 425–429.
Mitochondrial DNA content affects the fertilizability of human oocytes.Crossref | GoogleScholarGoogle Scholar |

Shoubridge, E. A., and Wai, T. (2007). Mitochondrial DNA and the mammalian oocyte. Curr. Top. Dev. Biol. 77, 87–111.
Mitochondrial DNA and the mammalian oocyte.Crossref | GoogleScholarGoogle Scholar |

Song, W. H., Ballard, J. W., Yi, Y. J., and Sutovsky, P. (2014). Regulation of mitochondrial genome inheritance by autophagy and ubiquitin-proteasome system: implications for health, fitness, and fertility. BioMed Res. Int. 2014, 981867.
Regulation of mitochondrial genome inheritance by autophagy and ubiquitin-proteasome system: implications for health, fitness, and fertility.Crossref | GoogleScholarGoogle Scholar |

Spikings, E. C., Alderson, J., and St John, J. C. (2007). Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development. Biol. Reprod. 76, 327–335.
Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development.Crossref | GoogleScholarGoogle Scholar |

St. John, J. C., Facucho-Oliveira, J., Jiang, Y., Kelly, R., and Salah, R. (2010). Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells. Hum. Reprod. Update 16, 488–509.
Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells.Crossref | GoogleScholarGoogle Scholar |

Tervit, H. R., Whittingham, D. G., and Rowson, L. E. (1972). Successful culture in vitro of sheep and cattle ova. J. Reprod. Fertil. 30, 493–497.
Successful culture in vitro of sheep and cattle ova.Crossref | GoogleScholarGoogle Scholar |

Tharasanit, T., Colenbrander, B., and Stout, T. A. (2005). Effect of cryopreservation on the cellular integrity of equine embryos. Reproduction 129, 789–798.
Effect of cryopreservation on the cellular integrity of equine embryos.Crossref | GoogleScholarGoogle Scholar |

Thundathil, J., Filion, F., and Smith, L. C. (2005). Molecular control of mitochondrial function in preimplantation mouse embryos. Mol. Reprod. Dev. 71, 405–413.
Molecular control of mitochondrial function in preimplantation mouse embryos.Crossref | GoogleScholarGoogle Scholar |

Tremoleda, J. L., Stout, T. A., Lagutina, I., Lazzari, G., Bevers, M. M., Colenbrander, B., and Galli, C. (2003). Effects of in vitro production on horse embryo morphology, cytoskeletal characteristics, and blastocyst capsule formation. Biol. Reprod. 69, 1895–1906.
Effects of in vitro production on horse embryo morphology, cytoskeletal characteristics, and blastocyst capsule formation.Crossref | GoogleScholarGoogle Scholar |

Wai, T., Ao, A., Zhang, X., Cyr, D., Dufort, D., and Shoubridge, E. A. (2010). The role of mitochondrial DNA copy number in mammalian fertility. Biol. Reprod. 83, 52–62.
The role of mitochondrial DNA copy number in mammalian fertility.Crossref | GoogleScholarGoogle Scholar |

Wang, L. Y., Wang, D. H., Zou, X. Y., and Xu, C. M. (2009). Mitochondrial functions on oocytes and preimplantation embryos. J. Zhejiang Univ. Sci. B 10, 483–492.
Mitochondrial functions on oocytes and preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |

Wilding, M., Dale, B., Marino, M., di Matteo, L., Alviggi, C., Pisaturo, M. L., Lombardi, L., and De Placido, G. (2001). Mitochondrial aggregation patterns and activity in human oocytes and preimplantation embryos. Hum. Reprod. 16, 909–917.
Mitochondrial aggregation patterns and activity in human oocytes and preimplantation embryos.Crossref | GoogleScholarGoogle Scholar |