The role of RhoA in the germinal vesicle breakdown of mouse oocytes
The role of RhoA in the germinal vesicle breakdown of mouse oocytes
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DOI：
10.1006/bbrc.2000.3052
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发表时间：
2000-07-14
影响因子：
3.1
通讯作者：
Ha, KS
中科院分区：
文献类型：
--
作者：
Cheon, YP;Kim, SW;Ha, KS
我们研究了RhoA在小鼠卵母细胞生殖泡破裂（GVBD）中的新作用。首先，RhoA通过免疫染色和ADP-核糖基化在生殖囊泡（GV）阶段卵母细胞中鉴定。RhoA主要定位于卵浆区，而在生殖泡中很少检测到。卵母细胞提取物与C3转移酶孵育诱导了约25 kDa的ADP-核糖基化。GV期卵母细胞在培养液中孵育分别诱导了约78%和87%的卵母细胞在1和3 h自发成熟为GVBD。在GV期卵母细胞中微量注射C3转移酶可显著抑制GVBD，在1和3 h时GVBD分别为29%和49%。为探讨活性氧（ROS）在卵母细胞成熟过程中的作用，用ROS特异性荧光染料H（2）DCFDA测定了GV期卵母细胞成熟过程中卵母细胞内ROS的水平。6 h时，微注射C3转移酶可抑制卵母细胞内ROS的产生。与ROS清除剂，N-乙酰-L-半胱氨酸和过氧化氢酶孵育，阻止ROS的增加。ROS清除剂也显著抑制GVBD，C3转移酶也是如此。因此，有人提出RhoA可能通过小鼠卵母细胞中ROS的产生参与GVBD，（C）2000 Academic Press.
We have investigated a new role of RhoA in the germinal vesicle breakdown (GVBD) of mouse oocytes. First, RhoA was identified by immunostaining and ADP-ribosylation in germinal vesicle (GV) stage-oocytes. RhoA was mainly localized in the ooplasmic area, but rarely detected in germinal vesicle. Incubation of oocyte extract with C3 transferase induced a strong ADP-ribosylation at about 25 kDa, Incubation of GV-stage oocytes in culture medium induced the spontaneous maturation to GVBD by about 78 and 87% of total oocytes at 1 and 3 h, respectively, However, microinjection of C3 transferase into GV-stage oocytes significantly inhibited GVBD at 1 (GVBD = 29%) and 3 h (GVBD = 49%). To study the role of reactive oxygen species (ROS) in the oocyte maturation, the level of intra oocyte ROS was measured using a ROS-specific fluorescent dye H(2)DCFDA during the oocyte maturation, Spontaneous maturation of GV-stage oocytes induced a significant increase of ROS at 3 h by about twofold over the control level and then the increased level was maintained until. 6 h, However, microinjection of C3 transferase inhibited the production of intra-oocyte ROS. Incubation with ROS scavengers, N-acetyl-L-cysteine and catalase, blocked the ROS increase. The ROS scavengers also significantly inhibited GVBD, as did C3 transferase. Thus, it was proposed that RhoA was involved in the GVBD, possibly by the production of ROS in mouse oocytes, (C) 2000 Academic Press.