title: DNA methylation pattern in mouse oocytes and their in vitro fertilized early embryos: effect of oocyte vitrification.
abstract: This study was conducted to investigate the pattern of DNA methylation in vitrified-thawed mouse oocytes and their in vitro fertilized early embryos. Firstly, mouse oocytes at metaphase II (MII) stage of meiosis were allocated randomly into three groups: (1) untreated (control); (2) exposed to vitrification solution without being plunged into liquid nitrogen (toxicity); or (3) vitrified by open-pulled straw (OPS) method (vitrification). Oocytes from all three groups were fertilized subsequently in vitro. The level of DNA methylation in the MII oocytes and their early embryos was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Developmental rates to 2-cell embryos (62.28%) and blastocysts (43.68%) of the vitrified-thawed oocytes were lower (P < 0.01) than those of fresh oocytes (81.47%, 61.99%) and vitrification solution treated (79.20%, 60.04%) oocytes. DNA methylation (as reflected by 5-MeC fluorescence intensity) in the vitrification group was less (P < 0.01) for MII oocyte and 2- to 8-cell stages compared with that in the control and toxicity groups. Accordingly, a reduction in global genomic methylation due to vitrification of MII oocytes may result in compromised in vitro developmental potential in early mouse embryos.
authors.affiliation: Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, and State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, and State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Xiang-Wei
authors.initials: XW
authors.lastname: Fu
authors.affiliation: Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, and State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Li
authors.affiliation: Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, and State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Dian-Shuai
authors.initials: DS
authors.lastname: Yuan
authors.affiliation: Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, and State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Shi-En
authors.initials: SE
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D015925:Cryopreservation; D019175:DNA Methylation; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D051379:Mice; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D058989:Vitrification