The effect of laser-assisted hatching on the methylation and expression pattern of imprinted gene IGF2/H19 in mouse blas
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EMBRYO BIOLOGY
The effect of laser-assisted hatching on the methylation and expression pattern of imprinted gene IGF2/H19 in mouse blastocysts and offspring Peng Huo 1 & Kai Deng 2 & Lulu Wang 2 & Man Li 3 & Jun Yao 3 & Jianghua Le 3 & Xiaocan Lei 4 & Shun Zhang 3 Received: 19 June 2020 / Accepted: 7 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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.
Peng Huo and Kai Deng contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10815-020-01975-4) contains supplementary material, which is available to authorized users. * Xiaocan Lei [email protected] * Shun Zhang [email protected]
Jianghua Le [email protected] 1
School of Public and Health, Guilin Medical University, Guilin 541004, China
2
Lulu Wang [email protected]
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
3
Man Li [email protected]
Department of Reproductive Medical Center, The Affiliated Hospital of Guilin Medical University, Guilin 541001, China
4
Clinical Anatomy & Reproductive Medicine Application Institute, Department of Histology and Embryology, University of South China, Hengyang 421001,
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