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Oocyte Vitrification Reduces its Capability to Repair Sperm DNA Fragmentation and Impairs Embryonic Development

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Abstract

Oocytes play a crucial role in repairing sperm DNA damage, which can affect the next generation; however, certain factors can impair this ability. This study examined whether oocyte vitrification, a widely used method for fertility preservation, negatively affects repair ability. Male DBA/2 mice (n = 28) were injected with 101.60 µmol/100 g body weight of tert-Butyl hydroperoxide (tBHP) for 14 days to induce sperm DNA damage. Histological changes, sperm functions, and DNA fragmentation were assessed using the TUNEL assay. Cumulus-oocyte-complexes (COCs) of superovulated female DBA/2 mice (n = 28) were vitrified using the Cryotop method. Fresh and vitrified oocytes were then fertilized by tBHP-treated and untreated sperms, and subsequent embryonic development was monitored. Additionally, the expression of Mre11a, Rad51, Brca1, and Xrcc4 was assessed in resulting zygotes and blastocysts using real-time PCR. The sperm tBHP treatment reduced differentiated spermatogenic cells in the testicular tissue, sperm concentration, and motility, while increasing DNA fragmentation (P < 0.05). The fertilization rate was decreased in the tBHP-treated sperm–vitrified oocyte group (P < 0.05), and the two-cell rate diminished in tBHP-treated sperm–fresh and vitrified oocyte groups (P < 0.05). The four-cell to blastocyst rate decreased in the untreated sperm–vitrified oocyte and the tBHP-treated sperm–fresh and vitrified oocyte groups (P < 0.05), and the tBHP-treated sperm–vitrified oocyte groups had the lowest blastocyst rate. In zygotes, Brca1 was upregulated in the tBHP-treated sperm–vitrified oocyte group (P < 0.05). Also, in blastocysts, Rad51, Brca1, and Xrcc4 were significantly upregulated in the untreated sperm–vitrified oocytes group (P < 0.05). Damages to the oocyte due to vitrification can disrupt the repair of sperm DNA fragmentation and consequently impair the embryo development.

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Acknowledgements

The authors thank all colleagues in the embryology lab, sperm biology lab, animal science lab, molecular lab, and histology lab of Royan Institute.

Funding

This project (96000285) was funded by Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine.

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Authors and Affiliations

  1. Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Niloofar Khajedehi & Hamid Gourabi

  2. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

    Niloofar Khajedehi & Rouhollah Fathi

  3. Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Vahid Akbarinejad

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  1. Niloofar Khajedehi
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Contributions

N.K. conducted all of the experimental work and drafted the manuscript. R.F. designed and supervised the study and interpreted results, mentioned fundamental advice to develop the experiment, and reviewed the manuscript. V.A. ran a statistical analysis, mentioned fundamental advice to develop the experiment, and reviewed the manuscript. H.G. proposed the hypothesis, designed and supervised the study and interpreted results, mentioned fundamental advice to develop the experiment, and reviewed the manuscript. All authors contributed to the report.

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Correspondence to Rouhollah Fathi or Hamid Gourabi.

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Khajedehi, N., Fathi, R., Akbarinejad, V. et al. Oocyte Vitrification Reduces its Capability to Repair Sperm DNA Fragmentation and Impairs Embryonic Development. Reprod. Sci. (2023). https://doi.org/10.1007/s43032-023-01419-1

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