title: The protective role of antifreeze protein 3 on the structure and function of mature mouse oocytes in vitrification.
abstract: Several studies have reported the oocyte damage in mice during vitrification; however, little has been known about the protective role that antifreeze protein 3 (Afp3) plays on their cellular structure and function during vitrification. In order to observe the extracellular cryo-protective role of Afp3, four groups were divided randomly. The observations were made for changes in cytoskeleton, expression of the related genes before and after vitrification, and also for changes in the in vitro developmental potential of oocytes. The outcomes were as follows: (i) microtubules, actin filaments and chromosomal integrity were more intact in the vitrification group supplemented with additional Afp3 compared to the vitrification group. In the fresh control group and the group with additional cryoprotectant containing ethylene glycol (EG), dimethyl sulfoxide (Me2SO) and sucrose, the organelles were more intact than the other two vitrification groups. (ii) Real-time PCR analysis revealed that the relative quantification of mitotic arrest deficient 2 (Mad2) and centromere protein E (Cenp-e) were significantly higher in the vitrification group with additional Afp3, the fresh control group and the one group with additional cryoprotectant, in comparison to the vitrification group. On the contrary, the expression of cold inducible RNA-binding protein (Cirbp) and kinesin-5 motor protein (Eg5) were up-regulated in the vitrification group compared to the remaining groups. (iii) The fertilization rate and the recovery rate in the fresh control group and the group with additional cryoprotectant were higher than the other two vitrification groups; furthermore, the recovery rate and the fertilization rate in the vitrification group with Afp3 were higher than the vitrification group. However, the blastocyst formation rate in all the four groups showed no statistical significance. In conclusion, Afp3 plays a positive role in the structure and function of mice oocytes in vitrification.
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wen
authors.affiliation: Center for Reproductive Medicine, Zaozhuang Maternal and Children Health Hospital, Zaozhuang 370400, Shandong, People's Republic of China.
authors.forename: Shuqin
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China.
authors.forename: Lan
authors.initials: L
authors.lastname: Chao
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China.
authors.forename: Hongling
authors.initials: H
authors.lastname: Yu
authors.affiliation: Academy of Medical Sciences, Jinan 250001, Shandong, People's Republic of China.
authors.forename: Changzheng
authors.initials: C
authors.lastname: Song
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China.
authors.forename: Honglei
authors.initials: H
authors.lastname: Chen
authors.affiliation: Center for Reproductive Medicine, Qilu Hospital of Shandong University, Jinan 250012, Shandong, People's Republic of China. Electronic address: dxh0122@163.com.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Deng
mesh_terms: D000818:Animals; D021343:Antifreeze Proteins, Type III / Q000378:metabolism*; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D008297:Male; D051379:Mice; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D058989:Vitrification
chemical_list: D021343:Antifreeze Proteins, Type III; D003451:Cryoprotective Agents
keywords: Antifreeze protein 3; Cytoskeleton; Mature oocyte; Vitrification