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S-adenosyl-homocysteine treatment of adult female fibroblasts alters X chromosome inactivation and improves in vitro embryo development after somatic cell nuclear transfer

B Jeon, Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada
G Coppola, Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada
S Perrault, Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada
G Rho, College of Veterinary Medicine, Gyeongsang National University, Jinju, Korea, Republic of
D Betts, Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada
W King, Biomedical Sciences, University of Guelph, Guelph, Ontario, NiG 2W1, Canada

Correspondence: Wa King, Email: waking{at}uoguelph.ca

Abstract

The poor outcome of somatic cell nuclear transfer (SCNT) is thought to be a consequence of incomplete reprogramming of the donor cell. The objective of this study was to investigate the effects of S-adenoysl homocysteine (SAH) treatment, a DNA demethylation agent, on DNA methylation levels and X-chromosome inactivation (XCI) status of bovine female fibroblast donor cell and the subsequent impact on developmental potential after SCNT. Compared with non treated controls, cells treated with SAH revealed i) significantly (P<0.05) reduced global DNA methylation, ii) significantly (1.5-fold) increased telomerase activity, iii) diminished distribution signals of methylated histones H3-3mK9 and H3-3mK27 on the presumptive inactive X chromosome (Xi) iv) alteration in the replication pattern of the Xi, v) elevation of transcript levels for X-chromosome linked genes, ANT3, HPRT, MeCP2, XIAP, XIST and HPRT. SCNT embryos produced with SAH treated donor cells compared with those derived from untreated donor cells revealed i) similar cleavage frequencies ii) significant elevation in the frequencies of development of cleaved embryos to hatched blastocyst stage iii) 1.5 fold increase in telomerase activity. We concluded that SAH induces global DNA demethylation, which partially reactivates the Xi and that a hypomethylated genome may facilitate the nuclear reprogramming process.