Xue-Ming Zhao, Ph.D., Jing-Jing Ren, M.Sc., Wei-Hua Du, Ph.D., Hai-Sheng Hao, Ph.D., Dong Wang, Ph.D., Yan Liu, Ph.D., Tong Qin, Ph.D., Hua-Bin Zhu, Ph.D.
Vol 98, Issue 1 , Pages 222-227
To determine the effect of vitrification and 5-aza-2'-deoxycytidine (5-aza-dC) on the methylation levels of the putative imprinted control region (ICR) of H19 and H19 expression in bovine two-cell embryos and their derived blastocysts.
Experimental animal study.
Abattoir-derived bovine ovaries.
Vitrified two-cell embryos were cultured in vitro to blastocysts with 0.01 μM 5-aza-dC (5-aza-dC group) or without 5-aza-dC (vitrification group). Fresh embryos and their derived blastocysts were used as control.
Main Outcome Measure(s):
Putative ICR methylation of H19 was measured by bisulfate mutagenesis and sequencing, blastocyst development rate; total cell number were determined; and H19 expression was measured by real-time reverse transcriptase-polymerase chain reaction (PCR).
Vitrification significantly increased putative ICR methylation of H19 in two-cell embryos and their derived blastocysts; 5-aza-dC significantly reduced putative ICR methylation of H19 in vitrified two-cell embryos and their derived blastocysts. The H19 expression level was significantly higher in blastocysts from the 5-aza-dC group than the vitrification group. The blastocyst development rate and total cell number in the 5-aza-dC and vitrification groups were similar.
Putative ICR methylation levels of H19 significantly increased in vitrified two-cell embryos and their derived blastocysts; 5-aza-dC significantly reduced putative ICR methylation of H19 and increased H19 expression in blastocysts derived from vitrified two-cell embryos.
Read the full text at: http://www.fertstert.org/article/S0015-0282(12)00446-3/fulltext