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The effect of laser-assisted hatching on the methylation and expression pattern of imprinted gene IGF2/H19 in mouse blastocysts and offspring

  • Embryo Biology
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Abstract

Purpose

This study aimed to determine the effects of drilling and thinning treatment of laser-assisted hatching on the expression and methylation of imprinted gene IGF2/H19 in embryos and offspring.

Methods

The prehatching blastocysts with treatment of drilling or thinning, or control prehatching blastocysts, were transplanted in surrogate uteri. The DNA methylation of IGF2/H19 imprinting control region (ICR) and the expression of IGF2 and H19 were respectively evaluated using bisulfite conversion-mediated sequencing and real-time PCR.

Results

The drilling group showed a significant increase in the development rate of hatched blastocysts in comparison with the control and thinning group. DNA methylation level of IGF2/H19 ICR of hatched blastocysts in the thinning group was 27.33% in comparison with the 38.67% and 36% observed in the control and drilling group. The thinning treatment increased the DNA methylation level of IGF2/H19 ICR in the placenta in comparison with the control and drilling group. The drilling and thinning treatment decreased the expression level of H19 mRNA in prehatching and hatched blastocysts as well as placenta, while a significant increase in the expression level of H19 mRNA of offspring was observed in the thinning group. The thinning treatment increased the expression level of IGF2 mRNA of prehatching blastocysts and offspring and a significant decrease in placenta, while the drilling treatment resulted in a significant increase in the expression level of IGF2 mRNA of hatched blastocysts and placenta.

Conclusion

These observations suggested that drilling used for hatching of in vitro cultured mouse blastocysts may improve the production of offspring.

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Funding

The present study was supported by Guangxi Science Foundation (No. 2017GXNSFBA198189), the China National Natural Science Fund (No. 81960274, 81860733 and 81860285), Guangxi Bagui Scholar Program and the Key Project of Guangxi Natural Science Foundation (No. 2018GXNSFDA050003).

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Author notes

  1. Peng Huo and Kai Deng contributed equally to this work.

Authors and Affiliations

  1. School of Public and Health, Guilin Medical University, Guilin, 541004, China

    Peng Huo

  2. Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University, Nanning, China

    Kai Deng & Lulu Wang

  3. Department of Reproductive Medical Center, The Affiliated Hospital of Guilin Medical University, Guilin, 541001, China

    Man Li, Jun Yao, Jianghua Le & Shun Zhang

  4. Clinical Anatomy & Reproductive Medicine Application Institute, Department of Histology and Embryology, University of South China, Hengyang, 421001, China

    Xiaocan Lei

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  1. Peng Huo
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  4. Man Li
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Contributions

PH, XL, and SZ directed the study; PH, KD, and ML performed the experiments; JY and JL performed the data collection and statistical analysis; JL edited the language; all the authors approved the manuscript for submission.

Corresponding authors

Correspondence to Xiaocan Lei or Shun Zhang.

Ethics declarations

All mice were raised in the Experimental Animal Centre of Guilin under a good feeding condition. All procedures described within were reviewed and approved by the Experimental Animal Care and Use Committee of Guilin and were performed in accordance with the Guiding Principles for the Care and Use of Laboratory Animals.

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The authors declare that they have no conflict of interest.

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Huo, P., Deng, K., Wang, L. et al. The effect of laser-assisted hatching on the methylation and expression pattern of imprinted gene IGF2/H19 in mouse blastocysts and offspring. J Assist Reprod Genet 37, 3057–3067 (2020). https://doi.org/10.1007/s10815-020-01975-4

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