Parthenogenetic activation of mouse oocytes by strontium chloride: A search for the best conditions
Parthenogenetic activation of mouse oocytes by strontium chloride: A search for the best conditions
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DOI：
10.1016/j.theriogenology.2005.03.002
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发表时间：
2005-09-15
期刊：
影响因子：
2.8
通讯作者：
Tan, JH
中科院分区：
文献类型：
--
作者：
Ma, SF;Liu, XY;Tan, JH
在核移植等实验中，锶已成功地用于诱导小鼠卵母细胞活化，但最佳处理条件尚未得到系统研究。当用10 mM SrCl 2处理无卵丘卵母细胞0.5-5 h时，激活率（88.4 +/- 4.1至91.2 +/- 2.7%）没有差异（平均S.E.; P > 0.2），但囊胚形成率（57.3 ± 3.5%）和每个囊胚细胞数（45.0 ± 2.4）在处理2.5 h后最高。在1-20 mM SrCl_2处理2.5h时，10 mM SrCl_2处理组的激活率和囊胚细胞数显著高于其他处理组（P < 0.02）。无Ca ~（2+）的Sr ~（2+）培养基的激活率最高，但在含Ca ~（2+）的培养基中激活率较低（37.7 ± 1.6%）。激活率是相同的，无论是否存在细胞松弛素B（CB）在激活培养基中，但囊胚形成率较高（P < 0.001）在CB的存在。只有70%的卵丘封闭的卵母细胞被激活，10%的囊胚后，暴露于1.6 mM的SrCl 2，许多裂解，增加强度的Sr 2+治疗。CB在SrCl 2培养基中的存在显着减少卵丘封闭的卵母细胞的裂解。当用作活化培养基时，培养基M16和CZB没有差异。在hCG后13 h收集的卵母细胞中，仅10.5%被Sr 2+单独处理激活，囊胚形成率为34%，但当它们进一步用6-二甲基氨基嘌呤（6-DMAP）处理时，激活率和囊胚形成率分别增加（P < 0.001）至94%和60%。孤雌生殖胚胎的总细胞数和ICM细胞数均低于体内受精胚胎（P < 0.001）。总之，SrCl 2处理的浓度和持续时间以及激活培养基和卵丘细胞中CB的存在或不存在对小鼠卵母细胞的激活和发育有显着影响。为了获得最佳的激活和发育，hCG后18小时收集的无卵丘卵母细胞应在补充有5 μ g/mL CB的无Ca 2+培养基中用10 mM SrCl 2处理2.5小时。(c)2005年爱思唯尔公司All rights reserved.
Strontium has been successfully used to induce activation of mouse oocytes in nuclear transfer and other experiments, but the optimum treatment conditions have not been studied systematically. When cumulus-free oocytes were treated with 10 mM SrCl2 for 0.5-5 h, activation rates (88.4 +/- 4.1 to 91.2 +/- 2.7%) did not differ (mean S.E.; P > 0.2), but rate of blastulation (57.3 +/- 3.5%) and cell number per blastocyst (45.0 +/- 2.4) were the highest after treatment for 2.5 h. When treated with 1-20 mM SrCl2 for 2.5 h, the activation rate and cell number per blastocyst were higher (P < 0.02) after 10 mM SrCl2 treatment than other treatments. The best activation and development were obtained :with Ca2+-free Sr2+ medium, but the activation rate was low (37.7 +/- 1.6%) in Ca2+-containing medium. Activation rates were the same, regardless of the presence or absence of cytochalasin B (CB) in the activating medium, but the blastulation rate was higher(P < 0.001) in the presence of CB. Only 70% of the cumulus-enclosed oocytes were activated and 10% blastulated after a 10 min exposure to 1.6 mM SrCl2, and many lysed, with increased intensity of Sr2+ treatment. The presence of CB in SrCl2 medium markedly reduced lysis of cumulus-enclosed oocytes. Media M16 and CZB did not differ when used as activating media. Only 10.5% of the oocytes collected 13 h post hCG were activated by Sr2+ treatment alone, with 34% blastulating, but rates of activation and blastulation increased (P < 0.001) to 94 and 60%, respectively, when they were further treated with 6-dimethylaminopurine (6-DMAP). The total and ICM cell numbers were less (P < 0.001) in parthenotes than in the in vivo fertilized embryos. In conclusion, the concentration and duration of SrCl2 treatment and the presence or absence of CB in activating medium and cumulus cells had marked effects on mouse oocyte activation and development. To obtain the best activation and development, cumulus-free oocytes collected 18 h post hCG should be treated for 2.5 h with 10 mM SrCl2 in Ca2+ free medium supplemented with 5 mu g/mL of CB. (c) 2005 Elsevier Inc. All rights reserved.