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Published in: Journal of Assisted Reproduction and Genetics 6/2018

Open Access 01-06-2018 | Embryo Biology

Low-dose irradiation of mouse embryos increases Smad-p21 pathway activity and preserves pluripotency

Authors: Masami Hayashi, Kayo Yoshida, Kohei Kitada, Akane Kizu, Daisuke Tachibana, Mitsuru Fukui, Takashi Morita, Masayasu Koyama

Published in: Journal of Assisted Reproduction and Genetics | Issue 6/2018

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Abstract

Purpose

To study the outcomes of mouse preimplantation embryos irradiated with low doses of X-rays (≤ 1 Gy) and investigate apoptosis and pluripotency of the irradiated embryos.

Methods

Mouse embryos at the 2-cell stage were collected for in vitro culture. After reaching the 8-cell stage, embryos were irradiated with various low doses of X-rays (0–1 Gy). Blastocysts with a normal appearance were transferred into a pseudopregnant uterus. The developmental rate to blastocysts and the survival rate following embryo transfer were examined. Expression levels of p21, Smad2, Foxo1, Cdx2, Oct4, and Nanog genes were measured by RT-PCR. Apoptotic cells in mouse blastocysts were examined immunofluorescently by staining for cleaved caspase-3.

Results

More than 90% of non-irradiated and low-dose X-ray-irradiated preimplantation embryos developed to morphologically normal blastocysts that could be implanted and survive in the uterus. However, embryos irradiated with X-rays had more apoptotic cells in a dose-dependent manner. Expression of p21, Smad2, and Foxo1 genes in X-ray-irradiated embryos was increased significantly, while expression of Cdx2, Oct4, and Nanog genes was maintained in comparison with non-irradiated embryos.

Conclusions

Although irradiated embryos contained apoptotic cells, the low doses of irradiation did not disturb development of 8-cell stage embryos to blastocysts or their survival in utero. The underlying mechanisms might involve anti-apoptotic systems, including the Smad-p21 pathway, and preservation of pluripotency.
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Metadata
Title
Low-dose irradiation of mouse embryos increases Smad-p21 pathway activity and preserves pluripotency
Authors
Masami Hayashi
Kayo Yoshida
Kohei Kitada
Akane Kizu
Daisuke Tachibana
Mitsuru Fukui
Takashi Morita
Masayasu Koyama
Publication date
01-06-2018
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 6/2018
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-018-1156-y

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