title: Vitrification transiently alters Oct-4, Bcl2 and P53 expression in mouse morulae but does not affect embryo development in vitro.
abstract: This study was conducted to determine the impact of vitrification on the expression of genes regulating pluripotency and apoptosis in mouse morulae. The morulae were randomly allocated into three groups: (1) untreated (control), (2) exposed to vitrification solution without freezing (toxicity), or (3) vitrified by open-pulled straw method (vitrification). In vitro development was evaluated by morphology and assessed by the blastocyst rate and the blastocyst total cell number. Gene expression in morulae and blastocysts was assessed by quantitative Real Time-PCR (qRT-PCR) and western blot. The results showed that at morulae stage, the POU class 5 homeobox1 (Oct-4) and B-cell lymphoma2 (Bcl2) mRNA levels of vitrification group were significantly lower (P < 0.05) than those of control. Strikingly, the p53 mRNA level was significantly higher in vitrification group. However, the Oct-4, Bcl2 and p53 mRNA levels in mouse blastocysts were not statistically different. Furthermore, western blot results showed that there was no significant difference in Oct-4, Bcl2 and p53 expression at protein level in mouse morulae among three groups. Additionally, the blastocyst rate (96.67%-100.00%) and the average cell number of blastocysts (89.67-92.33) were similar between all groups. The data demonstrate that vitrification transiently changes the mRNA expression of several key genes in mouse morulae regulating early embryo development but does not affect embryo developmental potential in vitro.
authors: Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, PR China. Electronic address: zguangbin@sicau.edu.cn.
authors: Guangbin
authors: G
authors: Zhou
authors: College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China; College of Animal Science and Technology, Southwest University, Chongqing 400715, PR China.
authors: Yan
authors: Y
authors: Zeng
authors: Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, PR China.
authors: Jiang
authors: J
authors: Guo
authors: Department of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, UT, USA.
authors: Qinggang
authors: Q
authors: Meng
authors: State Key Laboratory of AgroBiotechnology, China Agricultural University, Beijing 100193, PR China.
authors: Qingyong
authors: Q
authors: Meng
authors: Key Laboratory of Adaption and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, PR China.
authors: Gongxue
authors: G
authors: Jia
authors: College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors: Keren
authors: K
authors: Cheng
authors: Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, PR China.
authors: Changjun
authors: C
authors: Zeng
authors: Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, PR China.
authors: Ming
authors: M
authors: Zhang
authors: College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors: Guoshi
authors: G
authors: Liu
authors: College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors: Shi'en
authors: S
authors: Zhu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001755:Blastocyst / Q000502:physiology*; D002452:Cell Count; D015925:Cryopreservation / Q000379:methods; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005615:Freezing; D015870:Gene Expression; D051379:Mice; D009028:Morula / Q000502:physiology*; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis / Q000235:genetics*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics*; D058989:Vitrification
chemical_list: D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; C052617:Bcl2 protein, mouse
keywords: Apoptosis genes; Blastocyst rate; Mouse morulae; Oct-4; Vitrification