ROS和丁内酯-I抑制山羊卵母细胞的减数分裂  

Inhibiting the meiotic resumption of goat oocytes with roscovitine (ROS) and butyrolactoneI (BL-I)

作　　者：韩东[1] 吴延光[1] 兰国成[1] 韩正滨[1] 周佳勃[1] 谭景和[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《动物学报》2006年第2期342-348,共7页ACTA ZOOLOGICA SINICA

基　　金：国家自然科学基金项目(No.30430530);国家科技部"973"项目(No.G200016107)资助~~

摘　　要：本文研究了ROS(Roscovitine)和丁内酯-I(ButyrolactoneI,BL-I)两种细胞周期依赖性激酶抑制剂对山羊卵母细胞减数分裂恢复的抑制作用,并研究了抑制对卵母细胞成熟、激活和发育的影响。结果表明:ROS和BL-I对山羊卵母细胞减数分裂恢复的抑制作用具有浓度依赖性;200μmol/LROS、100μmol/LBL-I、100μmol/LROS+6·25μmol/LBL-I和50μmol/LROS+25μmol/LBL-I都能有效抑制山羊卵母细胞减数分裂的恢复,24h的抑制率分别为78·4%、80·9%、80·3%和77·8%。用ROS和BL-I抑制24h后转为正常培养24h,各处理组卵母细胞的成熟率(分别为81·3%、81·9%、83·2%和85·2%)与对照组(83·0%)无显著差异;成熟卵母细胞的化学激活率分别为93·3%、96·2%、92·5%和90·5%,与对照组(97·8%)无显著差异。然而,抑制处理后卵母细胞的卵裂率和桑椹胚率降低,未能发育到囊胚。ROS和BL-I抑制山羊卵丘扩展,并且转为正常培养后卵丘不能再扩展。ROS和BL-I能够浓度依赖性地抑制山羊卵母细胞减数分裂,二者既可单独,又可降低浓度联合使用,但抑制山羊卵母细胞的浓度远高于牛和猪卵母细胞的;ROS和BL-I抑制24h不影响山羊卵母细胞的成熟和激活能力,但影响卵母细胞的卵丘扩展和胚胎发育能力。因此,山羊卵母细胞减数分裂调控可能比它动物更精细[动物学报52(2):342-348,2006]。In vitro, when oocytes are removed from follicles, they accomplish nuclear maturation quickly with insufficient cytoplastic maturity, and this may lead to decreased developmental competence of IVM cocytes. Attempts have been made to prevent precocious meiotic resumption of oocytes and allow for adequate cytoplastic maturation in vitro. However, no such attempts were reported in goats. In this study, ROS and BL-Ⅰ, two specific and potent inhibitors of maturation-pro- moting factor （MPF） activity, were used to block germinal vesicle breakdown （GVBD） of goat oocytes, and their effects on cocyte maturation, activation and embryo development were studied. The results showed that both drugs blocked GV-BD of goat oocytes efficiently but their action was dose-dependent. Regimens of 200 μmol/L ROS, 100 μmol/L BL-Ⅰ, 100 μmol/L ROS ＋ 6.25 μmol/L BL-Ⅰ and 50 μmol/L ROS ＋ 25 μmol/L BL-Ⅰ blocked, respectively, 78.4%, 80.9%, 80.3% and 77.8% of oocytes in germinal vesicle （GV） stage during a 24-h culture period. Following a further 24 h culture in maturation medium, almost all （81.3%, 81.9%, 83.2% and 85.2% ） of the inhibited oocytes resumed meiosis and developed to the metaphase Ⅱ （MⅡ ） stage. When oocytes matured after inhibition were activated with ionomycin and 6-DMAP, activation rates in the 4 treatment groups were 93.3%, 96.2%, 92.5% and 90.5%, not different from that （97.8%） of control group. However, rates of cleavage and morulae of treated oocytes were lower than those of control oocytes. ROS and BL-Ⅰ inhibited cumulus expansion and the inhibitory effect was not reversible after maturation culture. In conclusion, both ROS and BL-Ⅰ inhibited meiotic resumption of goat oocytes in a dose-dependent manner and the dosage of ROS and BL-Ⅰ can be reduced to a big margin when used in combinations. Inhibition with ROS and BL-Ⅰ for 24 h did not compromise subsequent maturation and activation, but affected cumulus expansion and embryo development of goat oocytes .

关 键 词：山羊 卵母细胞 体外成熟 减数分裂 ROS丁内酯-Ⅰ 

分 类 号：Q813[生物学—生物工程]