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Seminal cell–free DNA as a potential marker for in vitro fertility of Nellore bulls

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

This study aimed to identify a marker for freezability and in vitro fertility of sperm samples before freezing.

Methods

Semen was collected from nine Nelore bulls; half of the ejaculate was used for seminal plasma cell–free DNA (cfDNA) quantification, and the other half was cryopreserved. Evaluation of sperm movement using computer-assisted semen analysis and plasma membrane integrity and stability, acrosomal integrity, apoptosis, and mitochondrial potential using flow cytometry were performed on fresh and frozen/thawed semen at 0, 3, 6, and 12 h after thawing. Frozen/thawed sperm was also used for in vitro embryo production. cfDNA was extracted from each bull, and the total DNA and number of cell-free mitochondrial DNA (cfmtDNA) copies were quantified. Semen from each animal was used for IVF, and cleavage, blastocyst formation, and cell counts were evaluated.

Results

Two groups were formed and compared based on the concentrations of cfDNA and cfmDNA present: low-cfDNA and high-cfDNA and low-cfmtDNA and high-cfmtDNA. Up to 12 h post-thawing, there were no differences between the groups in the majority of the sperm parameters evaluated. Cleavage, day 6 and 7 blastocyst rates, and the number of cells were higher in the high cfDNA group than in the low cfDNA group. Similar results were observed for cfmtDNA, except for the number of cells, which was similar between the groups.

Conclusion

The concentration of cfDNA and the relative number of copies of cfmtDNA in seminal plasma cannot predict the freezability of semen but can be used to predict in vitro embryo production.

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Acknowledgements

The authors would like to thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior —Brazil (CAPES) and Embrapa, Brazil. Margot Alves Nunes Dode and Maurício Machaim Franco were CNPq research fellows.

Funding

This study was financially supported by a grant from the Fundação de Apoio a Pesquisa do Distrito Federal (FAPDF — grant no. 019300001128).

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Authors and Affiliations

  1. Institute of Biology, University of Brasilia, Brasília, DF, Brazil

    Margot A. N. Dode, Natalia Capobianco & Nayara Ribeiro Kussano

  2. Laboratory of Animal Reproduction, Embrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil

    Margot A. N. Dode, Natalia Capobianco, Luna Nascimento Vargas, Nayara Ribeiro Kussano & Mauricio Machaim Franco

  3. Department of Animal Science, University of Guelph, Guelph, ON, Canada

    Bruna Mion

  4. School of Veterinary and Animal Science, Federal University of Goiás, Goiania, GO, Brazil

    José Felipe Spricigo

  5. School of Veterinary Medicine, Federal University of Uberlândia, Uberlandia, Minas Gerais, Brazil

    Mauricio Machaim Franco

  6. Institute of Biotechnology, Federal University of Uberlândia, Uberlandia, Minas Gerais, Brazil

    Mauricio Machaim Franco

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  1. Margot A. N. Dode
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Correspondence to Margot A. N. Dode.

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Dode, M.A.N., Capobianco, N., Vargas, L.N. et al. Seminal cell–free DNA as a potential marker for in vitro fertility of Nellore bulls. J Assist Reprod Genet 41, 1357–1370 (2024). https://doi.org/10.1007/s10815-024-03068-y

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