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Epigenetic effect of putrescine supplementation during in vitro maturation of oocytes on offspring in mice

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

Abstract

Purpose

To investigate the epigenetic safety of putrescine supplementation during in vitro maturation (IVM) to offspring.

Methods

Germinal vesicle oocytes retrieved from 12-week-old mice were randomly divided into two groups and cultured in IVM medium with or without 1 mmol/L putrescine for 16 h. Then, in vitro fertilization and embryo transplantation were conducted to produce the F1 offspring. The F1 mated with ordinary mice and bred the F2 offspring. The DNA methylation patterns in the brain and heart of F1 were investigated by reduced representation bisulfite sequencing. Imprinted gene expression levels of F1 oocytes were tested. The global methylation of F2 was examined by dot blot.

Results

The weight, organ coefficient, and histology were normal in the F1 and F2 offspring from the putrescine-treated oocytes. An overall methylation level of 31.23 to 32.53% was observed for all CpG sites in the brain and heart of the two groups. The DNA methylation patterns of the brain and heart in F1 were not altered in general, with subtle differences. The expression levels of imprinted genes including H19, Snrpn, Peg3, Igf2, and Igf2r did not statistically change. The global 5mC level of F2 was consistent with the control group.

Conclusion

Putrescine supplementation during IVM did not directly affect the development, health, and reproduction, and did not affect the genome and global epigenetics of mouse offspring derived from those oocytes. The transient putrescine treatment for improving oocyte maturation shows its long-term safety of genome and epigenetics in the offspring of mice.

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Data availability

The data underlying this article will be shared at reasonable request to the corresponding author.

Code availability

Not applicable.

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Acknowledgements

We thank Drs. Lingbo Cai and Yanqiu Hu at the Embryo Laboratory of our Center for their valuable help in the evaluation of oocyte quality and development potential, and Drs Lianju Qian and Chuyu Li in the cell culture.

Funding

This study was supported by grants from the National Nature and Science Foundation of China (81730041, 82171593) and the National Key Research and Development Program of China (2017YFC1001602, 2017YFC1001301).

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Authors and Affiliations

  1. State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, 210029, Jiangsu Province, China

    Chennan Shi, Jingyi Zhang, Zhengjie Yan, Li Gao, Chao Gao, Wei Wu, Jiayin Liu & Yugui Cui

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  1. Chennan Shi
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  2. Jingyi Zhang
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  8. Yugui Cui
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Contributions

Shi C., a graduate student of Professor Cui Y, performed the main experiments and wrote the first draft of the manuscript. Zhang J. and Yan Z. mainly helped in the embryonic experiments, Gao L. participated in the animal work, and Gao C. helped in the confocal and fluorescence work. Yan Z., Gao C., and Gao L. are technicians in Prof. Cui Lab. Wu W. participated in the putrescine analysis. Liu J. participated in the design and discussion of the manuscript. Cui Y. mainly designed the study, provided project funding, and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yugui Cui.

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All animal procedures and experiments were performed in accordance with the guidelines for animal care. The design and protocol of animal experiments were approved by the Animal Care Committee of Nanjing Medical University.

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The authors declare no competing interests.

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Shi, C., Zhang, J., Yan, Z. et al. Epigenetic effect of putrescine supplementation during in vitro maturation of oocytes on offspring in mice. J Assist Reprod Genet 39, 681–694 (2022). https://doi.org/10.1007/s10815-022-02448-6

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