Effect of mouse oocyte vitrification on mitochondrial membrane potential and distribution

Abstract

The effects of mouse oocyte vitrification on mitochondrial membrane potential and distribution were explored in this study. The collected mouse oocytes were randomly divided into vitrification and control groups. Ethylene glycol (EG) and dimethylsulphoxide (DMSO) were used as cryoprotectants in the vitrification group. The mitochondrial function and distribution in the oocytes were examined by using the fluorescent probes, JC-1 and Mito Tracker green. The results showed that the ratio of red to green fluorescence in mouse oocytes was significantly decreased after thawing in the vitrification group as compared with the control group (1.28 vs. 1.70, P<0.05). The percentage of polarized distribution of the mitochondria in oocytes was conspicuously reduced in the vitrification group when compared with the control group (31% vs. 63%, P<0.05). It was suggested that vitrification significantly affects the mitochondrial function and distribution in oocytes and reduces the potential of oocyte fertilization and embryo development.

Keywords

oocytes / vitrification / mitochondria / mice

References

[1] ChenC. Pregnancy after human oocyte cryopreservation. Lancet, 1986, 1(8486): 884-886

[2] DoveyS. Oocyte cryopreservation: advances and drawbacks. Minerva Ginecol, 2012, 64(6): 485-500

[3] BoriniA, BianchiV. Cryopreservation of mature and immature oocytes. Clin Obstet Gynecol, 2010, 53(4): 763-774

[4] LornageJ, SalleB. Ovarian and oocyte cryopreservation. Curr Opin Obstet Gynecol, 2007, 19(4): 390-394

[5] KuwayamaM, VajtaG, KatoO, et al.. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online, 2005, 11(3): 300-308

[6] AntinoriM, LicataE, DaniG, et al.. Cryotop vitrification of human oocytes results in high survival rate and healthy deliveries. Reprod Biomed Online, 2007, 14(1): 72-79

[7] HomburgR, van der VeenF, SilberSJ. Oocyte vitrification—women’s emancipation set in stone. Fertil Steril, 2009, 91(4Suppl): 1319-1320

[8] BoldtJ. Current results with slow freezing and vitrification of the human oocyte. Reprod Biomed Online, 2011, 23(3): 314-322

[9] WangY, ZhuG, LiX, et al.. Experimental study on meiotic spindles and chromosomes of mouse mature (M II) stage oocytes under laser scanning confocal microscopy. J Huazhong Univ Sci Technol [Med Sci], 2006, 26(3): 353-355

[10] Berthelot-RicouA, PerrinJ, di GiorgioC, et al.. Genotoxicity assessment of mouse oocytes by comet assay before vitrification and after warming with three vitrification protocols. Fertil Steril, 2013, 100(3): 882-888

[11] OuXH, LiS, WangZB, et al.. Maternal insulin resistance causes oxidative stress and mitochondrial dysfunction in mouse oocytes. Hum Reprod, 2012, 27(7): 2130-2145

[12] YongshengJ, XiaoyunM, XiaoliW, et al.. Antitumor activity of erythromycin on human neuroblastoma cell line (SH-SY5Y). J Huazhong Univ Sci Technol [Med Sci], 2011, 31(1): 33-38

[13] ReersM, SmileyST, Mottola-HartshornC, et al.. Mitochondrial membrane potential monitored by JC-1 dye. Methods Enzymol, 1995, 260: 406-417

[14] GosdenRG. Fertility preservation: definition, history, and prospect. Semin Reprod Med, 2009, 27(6): 433-437

[15] HerreroL, MartinezM, Garcia-VelascoJA. Current status of human oocyte and embryo cryopreservation. Curr Opin Obstet Gynecol, 2011, 23(4): 245-250

[16] EdgarDH, GookDA. A critical appraisal of cryopreservation (slow cooling versus vitrification) of human oocytes and embryos. Hum Reprod Update, 2012, 18(5): 536-554

[17] SmithGD, SerafiniPC, FioravantiJ, et al.. Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification. Fertil Steril, 2010, 94(6): 2088-2095

[18] NagaiS, MabuchiT, HirataS, et al.. Oocyte mitochondria: strategies to improve embryogenesis. Hum Cell, 2004, 17(4): 195-201

[19] GualtieriR, IaccarinoM, MolloV, et al.. Slow cooling of human oocytes: ultrastructural injuries and apoptotic status. Fertil Steril, 2009, 91(4): 1023-1034

[20] Eichenlaub-RitterU, WieczorekM, LukeS, et al.. Age related changes in mitochondrial function and new approaches to study redox regulation in mammalian oocytes in response to age or maturation conditions. Mitochondrion, 2011, 11(5): 783-796

[21] BreviniTA, CilloF, AntoniniS, et al.. Cytoplasmic remodelling and the acquisition of developmental competence in pig oocytes. Anim Reprod Sci, 2007, 98(1–2): 23-38

[22] LiR, AlbertiniDF. The road to maturation: somatic cell interaction and self-organization of the mammalian oocyte. Nat Rev Mol Cell Biol, 2013, 14(3): 141-152

[23] DengWP, RenZR. Mitochondrial and oocyte development. Yi Chuan (Chinese), 2007, 29(12): 1429-1433

[24] KatayamaM, ZhongZ, LaiL, et al.. Mitochondrial distribution and microtubule organization in fertilized and cloned porcine embryos: implications for developmental potential. Dev Biol, 2006, 299(1): 206-220

[25] LiY, FengHL, CaoYJ, et al.. Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro. Fertil Steril, 2006, 85(4): 827-832

[26] YinH, DuffyDM, GosdenRG. Comparative maturation of cynomolgus monkey oocytes in vivo and in vitro. Reprod Biol Endocrinol, 2006, 4: 14

[27] TornerH, AlmH, KanitzW, et al.. Effect of initial cumulus morphology on meiotic dynamic and status of mitochondria in horse oocytes during IVM. Reprod Domest Anim, 2007, 42(2): 176-183

[28] TakahashiT, HammettMF, ChoMS. Multifaceted freezing injury in human polymorphonuclear cells at high subfreezing temperatures. Cryobiology, 1985, 22(3): 215-236

[29] LovelockLE. The haemolysis of human red blood cells by freezing and thawing. Biochim Biophys Acta, 1953, 10: 414-426