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5-AZA-2′-deoxycytidine (5-AZA-CdR) leads to down-regulation of Dnmt1o and gene expression in preimplantation mouse embryos

Published online by Cambridge University Press:  01 May 2009

Jian-Ning Yu
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China. Institute of Animal Science, Jiangsu Academy of Agri-cultural Sciences, Nanjing, Jiangsu, P.R. China.
Chun-Yang Xue
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China.
Xu-Guang Wang
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China.
Fei Lin
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China.
Chun-Yi Liu
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China.
Fu-Zeng Lu
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, Jiangsu, P.R China.
Hong-Lin Liu*
College of Animal Science and Technology, Nanjing Agriculture University, No. 1 WeiGang Street, Nanjing, Jiangsu, P.R. China210095.
All correspondence to Hong-Lin Liu. College of Animal Science and Technology, Nanjing Agriculture University, No. 1 WeiGang Street, Nanjing, Jiangsu, P.R. China 210095. Tel: +86 25 84395106. e-mail:


5-AZA-2′-deoxycytidine (5-AZA-CdR) is a demethylating, teratogenic agent and a mutagen, which causes defects in the developing mouse and rat after implantation. Our previous data indicated that 5-AZA-CdR (0.2 and 1.0 μM) inhibited the development of mouse preimplantation embryos. Pronuclear embryos exposed to 5-AZA-CdR at the pronuclear stage were unable to form 8-cell embryos, while 2-cell-stage embryos exposed to 5-AZA-CdR only developed into uncompacted 8-cell-stage embryos. And there was no formation of blastocysts when 4-cell embryos cultured in 5-AZA-CdR. In our present study, we detected Dnmt1o protein and some developmental gene expression in order to find the reasons for the developmental arrest. Dnmt1o could not traffic to 8-cell nuclei as control when embryos were exposed to 5-AZA-CdR. Dnmt1o was in cytoplasm at 2-cell and 4-cell stages before and after treated with 5-AZA-CdR. Gene expression changes were also detected in this research. Our data indicated that connexin 31 (Cx31), connexin 43 (Cx43), connexin 45 (Cx45), E-cadherin (Cdh1) and β-catenin (Ctnnb1) were all downregulated by 5-AZA-CdR. Cx31, Cx43 and Cx45 are members of connexins family, which have a central role in gap junctions. Cdh1 and Ctnnb1 are necessary for the foundation of tight junctions. Therefore, developmental arrest induced by 5-AZA-CdR may be caused by the failure of Dnmt1o cytoplasmic–nuclear traffic and the down-regulation of developmental gene expression. Normal compaction and blastocoel cavitation need Dnmt1o traffic to 8-cell nuclei and the right gene expression, especially the correlative genes in gap junctions and tight junctions.

Research Article
Copyright © Cambridge University Press 2009

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