S-adenosylhomocysteine treatment of adult female fibroblasts alters X-chromosome inactivation and improves in vitro embryo development after somatic cell nuclear transfer.
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 treatment with S-adenosylhomocysteine (SAH) a DNA demethylation agent, on DNA methylation levels and X-chromosome inactivation status of bovine female fibroblast donor cells and the subsequent impact on developmental potential after SCNT. Compared with non-treated controls, the cells treated with SAH revealed (i) significantly (P<0.05) reduced global DNA methylation, (ii) significantly (approximately 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, and (v) elevation of transcript levels for X-chromosome linked genes, ANT3, 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, and (iii) 1.5-fold increase in telomerase activity. We concluded that SAH induces global DNA demethylation that partially reactivates the Xi, and that a hypomethylated genome may facilitate the nuclear reprogramming process.
Jeon
Byeong-Gyun
BG
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Coppola
Gianfranco
G
Perrault
Steven D
SD
Rho
Gyu-Jin
GJ
Betts
Dean H
DH
King
W Allan
WA
D044503:5-Methylcytosine / Q000032:analysis; D000818:Animals; D002417:Cattle; D065150:Cellular Reprogramming; D019175:DNA Methylation; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D006657:Histones / Q000032:analysis / Q000378:metabolism; D008677:Metaphase; D008856:Microscopy, Fluorescence; D053652:Nuclear Transfer Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012435:S-Adenosylhomocysteine / Q000494:pharmacology*; D019098:Telomerase / Q000378:metabolism; D014158:Transcription, Genetic; D049951:X Chromosome Inactivation / Q000187:drug effects*
D006657:Histones; D044503:5-Methylcytosine; D012435:S-Adenosylhomocysteine; D019098:Telomerase