Ye Xiao-Fang, Chen Shi-Bin, Wang Li-Qin, Zhao Yun-Cheng, Lv Xue-Feng, Liu Ming-Jun, Huang Jun-Cheng
Xinjiang Key Laboratory of Animal Biotechnology, Xinjiang Academy of Animal Sciences, Urumqi 830000, China.
Reprod Fertil Dev. 2010;22(8):1254-61. doi: 10.1071/RD10062.
The intracellular glutathione levels and developmental competence of aged oocytes after parthenogenetic activation, somatic cell nuclear transfer and intracytoplasmic sperm injection in the presence or absence of caffeine or dithiothreitol (DTT) were examined. The following results were found: (1) ovine oocytes were fully aged 30 h post-onset of maturation culture; (2) the appropriate concentrations of caffeine and DTT for oocyte culture were 5 mM and 1 mM, respectively; (3) when nuclear transfer-reconstructed embryos were treated with caffeine or DTT following fusion, no increase in the frequency of development to blastocyst was observed (P > 0.05), but the cell numbers of blastocysts increased (P < 0.05); (4) both caffeine and DTT increased the blastocyst formation rates of intracytoplasmic sperm-injected embryos (P < 0.05); (5) caffeine increased the glutathione content of aged oocytes (P < 0.05). The glutathione content of DTT-treated aged oocytes was higher than that of oocytes matured for 36 h (P < 0.05). In conclusion, caffeine and dithiothreitol delay oocyte ageing but only to a limited extent.
研究了在有或没有咖啡因或二硫苏糖醇(DTT)存在的情况下,孤雌激活、体细胞核移植和胞浆内精子注射后老龄卵母细胞的细胞内谷胱甘肽水平和发育能力。结果如下:(1)绵羊卵母细胞在成熟培养开始后30小时完全老化;(2)用于卵母细胞培养的咖啡因和DTT的适宜浓度分别为5 mM和1 mM;(3)当核移植重构胚胎在融合后用咖啡因或DTT处理时,观察到发育至囊胚的频率没有增加(P>0.05),但囊胚的细胞数量增加(P<0.05);(4)咖啡因和DTT均提高了胞浆内精子注射胚胎的囊胚形成率(P<0.05);(5)咖啡因增加了老龄卵母细胞的谷胱甘肽含量(P<0.05)。DTT处理的老龄卵母细胞的谷胱甘肽含量高于成熟36小时的卵母细胞(P<0.05)。总之,咖啡因和二硫苏糖醇可延缓卵母细胞老化,但程度有限。