Impaired active DNA demethylation in zygotes generated by round spermatid injection

Human Reproduction
Yoko Kakino KurotakiAtsuo Ogura

Abstract

Is the poor development of embryos generated from round spermatid injection (ROSI) in humans and animals associated with abnormal active DNA demethylation? A significant proportion of ROSI-derived embryos failed to undergo active DNA demethylation. Active DNA demethylation is initiated by the conversion of 5-methylcytosine (5mC) to 5-hydroxycytosine (5hmC) by the Tet3 enzyme. Active demethylation proceeds in a more pronounced manner in the male pronucleus than in the female one. Mouse zygotes generated by ICSI or ROSI were analyzed for active DNA methylation by quantification of 5mC and 5hmC using specific antibodies. Some ROSI-derived embryos were subjected to time-lapse imaging for DNA methylation levels and were transferred into recipient pseudo-pregnant female mice. In ICSI-derived embryos, the male:female pronucleus (M/F) ratio of 5mC immunostaining intensity was decreased while that of 5hmC was increased. However, a significant proportion of ROSI-derived embryos showed unchanged M/F ratios for 5mC and 5hmC even at the late zygotic period, indicating that they failed to undergo asymmetric active DNA demethylation. Consistent with this, some ROSI-derived embryos did not show preferential localization of Tet3 to the male pro...Continue Reading

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Citations

Oct 27, 2015·The Journal of Reproduction and Development·Pedro MoreiraAlfonso Gutiérrez-Adán
Feb 24, 2016·Systems Biology in Reproductive Medicine·Hoda RajabiMohammad Salehi
May 12, 2016·Theriogenology·Sebastian Canovas, Pablo Juan Ross
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Feb 20, 2018·Molecular Reproduction and Development·Narumi OgonukiAtsuo Ogura
Jun 20, 2019·Molecular Reproduction and Development·Narumi OgonukiAtsuo Ogura
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Oct 29, 2020·Andrology·Antoine ObletteChristine Rondanino
Apr 3, 2021·Frontiers in Cell and Developmental Biology·Haibo ZhuQi Zhou

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