巯基乙酸对爪蟾卵母细胞体外孤雌激活的影响及其机制研究

doi:10.3969/j.issn.1004-616x.2010.05.007

癌变·畸变·突变 ›› 2010, Vol. 22 ›› Issue (5): 356-360.doi: 10.3969/j.issn.1004-616x.2010.05.007

巯基乙酸对爪蟾卵母细胞体外孤雌激活的影响及其机制研究

任晓梅1,夏　蕾2,王　栋1,张　岭1,侯绍英1,赵　巍3,吴　坤1,

1．哈尔滨医科大学公共卫生学院营养与食品卫生教研室，黑龙江哈尔滨 150081； 2．哈尔滨医科大学附属第一医院生殖医学中心，黑龙江哈尔滨 150001； 3．哈尔滨市第一医院骨外二科，黑龙江哈尔滨 150010

收稿日期:2010-05-28 修回日期:2010-06-30 出版日期:2010-09-30 发布日期:2010-09-30
通讯作者: 吴　坤

Effects of thioglycolic acid on parthenogenetic activation of xenopus oocytes

REN Xiao-mei1， XIA Lei2， WANG Dong1， ZHANG Ling1，HOU Shao-ying1， ZHAO Wei3， WU Kun1,

1. Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical University, Harbin 150081;2. Department of Reproductive Medicine, The First Affiliated Hospital of Harbin Medical University, Harbin 150001; 3.The Second Department of Orthopaedics,the First Hospital in Harbin,Harbin 150010, China

Received:2010-05-28 Revised:2010-06-30 Online:2010-09-30 Published:2010-09-30
Contact: WU Kun

摘要/Abstract

摘要： 目的：探讨巯基乙酸(thioglycolic acid, TGA)对钙离子载体A23187诱导的非洲爪蟾卵母细胞体外孤雌激活的影响，探讨孤雌激活的机制。方法：体外孕酮诱导爪蟾卵母细胞成熟后，选取MII期细胞用不同浓度TGA(0、5、25、125 μg/ml)处理2 h，再用钙离子载体A23187诱导激活，在加入激活剂后不同时间(0、10、20、30、60 min)记录激活的卵母细胞数量，计算其激活率；加入激活剂后60 min收集卵母细胞，进行荧光染色观察原核形成情况；采用Western Blotting检测CyclinB2、Mos、p_ERK1/ERK1的表达。结果：不同浓度的TGA均降低非洲爪蟾卵母细胞的孤雌激活率，与对照组相比差异具有统计学意义(P<0.05)；激活后对照组卵母细胞在动物极中央形成一个圆形的受精斑，而TGA125 μg/ml处理后形成的受精斑形状不规则且周围毛糙不齐；荧光染色发现对照组细胞中有清晰的原核形成，而TGA处理组则未发现；Western Blotting 分析结果显示，经TGA处理可抑制CyclinB2、Mos、p_ERK1在孤雌激活时的降解。结论： TGA可以抑制钙离子载体A23187诱导的非洲爪蟾卵母细胞体外孤雌激活，其作用机制可能与干扰成熟促进因子(MPF)和丝裂原活化蛋白激酶(MAPK)在孤雌激活时的正常降解有关。

关键词: 巯基乙酸, 卵母细胞, 孤雌激活

Abstract: OBJECTIVE: To investigate the effects of thioglycolic acid(TGA) on calcium ionophore A23187_ induced parthenogenetic activation of xenopus oocytes and its possible mechanism. METHODS: Xenopus oocytes were pretreated with TGA in vitro at the doses of 0, 5, 25, 125 μg/ml, and then activated by A23187. During this process, the activation rates were noted. Oocytes were collected 1h after the addition of A23187 and stained with Hoechst 33258 to check the formation of pronucleus. RESULTS: Following TGA treatment, the black spot appeared on the animal pole, known as fertilization coat, which was not in good condition. TGA significantly inhibited the rate of parthenogenetic activation(P<0.05) and the formation of pronucleus. Through Western Blotting analysis, TGA inhibited the degradation of Mos, p_ERK1 and CyclinB2. CONCLUSION:TGA treatment could inhibit the parthenogenetic activation of xenopus oocytes induced by A23187, possibly due to the down_regulation of MPF and MAPK degradation.

Key words: thioglycolic acid, oocytes, parthenogenetic activation

任晓梅, 夏　蕾, 王　栋, 张　岭, 侯绍英, 赵　巍, 吴　坤, . 巯基乙酸对爪蟾卵母细胞体外孤雌激活的影响及其机制研究[J]. 癌变·畸变·突变, 2010, 22(5): 356-360.

REN Xiao-mei, XIA Lei, WANG Dong, ZHANG Ling, HOU Shao-ying, ZHAO Wei, WU Kun, . Effects of thioglycolic acid on parthenogenetic activation of xenopus oocytes[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2010, 22(5): 356-360.

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巯基乙酸对爪蟾卵母细胞体外孤雌激活的影响及其机制研究

Effects of thioglycolic acid on parthenogenetic activation of xenopus oocytes

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