Effect of 5-aza-2'-deoxycytidine on methylation of the putative imprinted control region of H19 during the in vitro development of vitrified bovine two-cell embryos

H19 imprinted control region methylation levels significantly increased in vitrified two-cell embryos and their derived blastocysts; 5-aza-dC significantly reduced H19 imprinted control region methylation and increased H19 expression in blastocysts derived from vitrified embryos.

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Authors

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

Abstract

Objective:

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.

Design:

Experimental animal study.

Setting:

Academic institution.

Animal(s):

Abattoir-derived bovine ovaries.

Intervention(s):

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).

Result(s):

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.

Conclusion(s):

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


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Fertility and Sterility® is an international journal for obstetricians, gynecologists, reproductive endocrinologists, urologists, basic scientists and others who treat and investigate problems of infertility and human reproductive disorders. The journal publishes juried original scientific articles in clinical and laboratory research relevant to reproductive endocrinology, urology, andrology, physiology, immunology, genetics, contraception, and menopause. Fertility and Sterility® encourages and supports meaningful basic and clinical research, and facilitates and promotes excellence in professional education, in the field of reproductive medicine.

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