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线粒体能量代谢和DNA复制对小鼠卵子体外成熟、受精及发育的影响
葛红山* 李晓和 张帆 陈华 习海涛 黄检英 朱春芳 吕杰强
解剖学报 ›› 2013, Vol. 44 ›› Issue (5) : 675-679.
线粒体能量代谢和DNA复制对小鼠卵子体外成熟、受精及发育的影响
Effects of mitochondrial metabolic activity and mitochondrial DNA replication on maturation, fertilization and developmental competence in vitro of rat oocytes
目的 探讨胞质线粒体的氧化磷酸化（OXPHOS）活性或线粒体DNA复制在卵子成熟、受精和胚胎发育过程中的作用。方法 通过在小鼠体外成熟培养液中引入不同浓度的羰基氰4-(三氟甲氧基)苯腙 (FCCP, 10 nmol/L和100nmol/L)或2′,3′-双脱氧胞苷(ddC,10μmol/L和100μmol/L)，抑制线粒体OXPHOS活性或线粒体DNA复制，统计分析各组卵子的体外生发泡破裂（GVBD）率、核成熟率、受精率及囊胚形成率，以分析线粒体功能抑制对卵子成熟、受精和胚胎发育的影响。 结果 线粒体OXPHOS活性和DNA复制功能在卵子和胚胎中所发挥的作用并不完全相同。FCCP抑制线粒体OXPHOS活性可显著降低卵子的核成熟率和囊胚形成率；但对卵子的GVBD的发生率和受精率无显著影响。而ddC抑制线粒体DNA复制不影响卵子的体外成熟和受精，但可显著抑制囊胚的形成。 结论 OXPHOS活性主要影响卵子成熟及胚胎发育；线粒体DNA复制则主要影响胚胎发育；而线粒体功能抑制不影响卵子的成熟启动和体外受精。
Objective To investigate the roles of mitochondrion on whether either mitochondrial DNA copy number or its metabolic capacity is an essential factor for both oocyte maturation and embryo development. Methods Both the metabolic inhibitor (FCCP) and the inhibitor of mitochondrial DNA replication(ddC) were introduced into the culture medium in order to analyze the affection of mitochondria rates of germinal vesicle breakdown. Nuclear maturation, fertilization and blastulation were assessedin vitro. Results The inhibiting mitochondrial DNA replication did not coincide with the mitochondrial metabolic capacity in oocytes. FCCP treatment significantly reduced the rate of nuclear maturation of oocytes and the development potential of embryos compared to untreated controls. Treatment of ddC did not affect maturationin vitro of oocytes, but impaired blastulation. Neither FCCP nor ddC treatment had an effect on the rate of fertilization. Conclusion Inhibiting mitochondrial metabolic activity during oocyte maturation has a negative impact on oocyte maturation and ensuing embryo developmental competence. Reduction in mitochondrial DNA copy number mainly affects embryonic development potential, but has little effect on oocyte maturation and fertilizationin vitro.
线粒体能量代谢 / 线粒体DNA / 卵子成熟 / 发育潜能 / 体外受精 / 小鼠
Mitochondrial energy metabolism / Mitochondrial DNA / Oocyte maturation / Developmental competence / Fertilization in vitro / Mouse
［1］Bentov Y, Esfandiari N, Burstein E,et al. The use of mitochondrial nutrients to improve the outcome of infertility treatment in older patients ［J］. Fertil Steril， 2010, 93(1):272-275.
［2］Liu Sh, Li Y, Gao X, et al. Chang of the mitochondrial distribution in human oocytes before and after in vitro maturation［J］. Acta Anatomica Sinica,2007,38(5):593-596.(in Chinese)
刘姗, 李媛, 高选,等. 人卵母细胞体外成熟前后线粒体分布的变化［J］ 解剖学报,2007,38(5):593-596.
［3］Van Blerkom J, Davis P. Mitochondrial signaling and fertilization［J］. Mol Hum Reprod,2007, 13(11):759-770.
［4］Stojkovic M, Machado SA, Stojkovic P, et al. Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and afterin vitro maturation: correlation with morphological criteria and developmental capacity afterin vitro fertilization and culture［J］. Biol Reprod,2001, 64(3):904-909.
［5］Reynier P, May-Panloup P, Chretien M, et al. Mitochondrial DNA content affects the fertilizability of human oocytes［J］. Mol Hum Reprod, 2001, 7(5):425-429.
［6］Van Blerkom J, Davis PW, Lee J. ATP content of human oocytes and developmental potential and outcome afterin vitro fertilization and embryo transfer［J］. Hum Reprod, 1995, 10(2):415-424.
［7］Spikings EC, Alderson J, John JCS. Regulated mitochondrial DNA replication during oocyte maturation is essential for successful porcine embryonic development［J］. Biol Reprod, 2007, 76(2):327-335.
［8］Santos TA, El Shourbagy S, St John JC. Mitochondrial content reflects oocyte variability and fertilization outcome［J］. Fertil Steril, 2006, 85(3):584-591.
［9］Zeng H, Yeung WSB, Cheung MPL, et al.In vitro-matured rat oocytes have low mitochondrial deoxyribonucleic acid and adenosine triphosphate contents and have abnormal mitochondrial redistribution［J］. Fertil Steril,2009, 91(3):900-907.
［10］Liu S, Li Y, Gao X, et al. Changes in the distribution of mitochondria before and after in vitro maturation of human oocytes and the effect of in vitro maturation on mitochondria distribution［J］. Fertil Steril, 2010, 93(5):1550-1555.
［11］Chan C, Liu V, Lau E,et al. Mitochondrial DNA content and 4977 bp deletion in unfertilized oocytes［J］. Mol Hum Reprod, 2005, 11(12):843-846.
［12］Acton B, Jurisicova A, Jurisica I, et al. Alterations in mitochondrial membrane potential during preimplantation stages of mouse and human embryo development［J］. Mol Hum Reprod, 2004, 10(1):23-32.
［13］Van Blerkom J, Davis P. Mitochondrial signaling and fertilization［J］. Mol Hum Reprod, 2007, 13(11):759-770.
［14］Thouas GA, Trounson AO, Wolvetang EJ, et al. Mitochondrial dysfunction in mouse oocytes results in preimplantation embryo arrest in vitro［J］. Biol Reprod, 2004, 71(6):1936-1942.
［15］Pikó L, Taylor KD. Amounts of mitochondrial DNA and abundance of some mitochondrial gene transcripts in early mouse embryos［J］. Dev Biol,1987, 123(2):364-374.
［16］El Shourbagy SH, Spikings EC, Freitas M,et al. Mitochondria directly influence fertilisation outcome in the pig［J］. Reproduction, 2006, 131(2):233-245.
［17］Thouas GA, Trounson AO, Jones GM. Developmental effects of sublethal mitochondrial injury in mouse oocytes［J］. Biol Reprod, 2006, 74(5):969-977.
［18］Steuerwald N, Barritt JA, Adler R, et al. Quantification of mtDNA in single oocytes, polar bodies and subcellular components by real-time rapid cycle fluorescence monitored PCR［J］. Zygote, 2000, 8(3):209-215.
［19］Wai T, Ao A, Zhang X, et al. The role of mitochondrial DNA copy number in mammalian fertility［J］. Biol Reprod, 2010, 83(1):52-62.
［20］Zeng H, Ren Z, Yeung WSB, et al. Low mitochondrial DNA and ATP contents contribute to the absence of birefringent spindle imaged with PolScope in in vitromatured human oocytes［J］. Hum Reprod,2007, 22(6):1681-1686.
国家863计划基金资助项目
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