Improvement in the in vitro maturation rate of porcine oocytes vitrified at the germinal vesicle stage by treatment with a mitochondrial permeability transition inhibitor
Improvement in the in vitro maturation rate of porcine oocytes vitrified at the germinal vesicle stage by treatment with a mitochondrial permeability transition inhibitor
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DOI：
10.1016/j.cryobiol.2008.09.008
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发表时间：
2008-12-01
期刊：
影响因子：
2.7
通讯作者：
Watanabe, Tornornasa
中科院分区：
文献类型：
--
作者：
Nakagawa, Shoma;Yoneda, Akihiro;Watanabe, Tornornasa
本研究通过形态学观察和体外成熟（IVM）方法，研究了线粒体通透性转换（MPT）抑制剂、caspase活性抑制剂、胞内Ca 2+螯合剂和线粒体Ca 2+单向转运体对猪玻璃化冷冻胚泡（GV）卵母细胞存活率的影响。玻璃化冷冻GV期卵母细胞在添加MPT抑制剂环孢菌素A（CsA）的IVM培养液中成熟时，其存活率和IVM率（分别为36.1%和26.8%）显著高于其他玻璃化冷冻组（分别为10.3-12.3%和6.2- 10.3%）（P < 0.05）。然而，半胱天冬酶抑制剂Z-VAD-fastrin（Z-VAD）并没有提高存活率和IVM率（分别为11.7-21.6%和8.5- 155%）。当IVM培养液中加入胞内钙离子螯合剂BAPTA-AM时，GV期卵母细胞玻璃化冷冻后的存活率和IVM率（分别为34.5-36.2%和25.0- 26.9%）显著高于不加玻璃化冷冻组（分别为17.2-24.2%和12.9- 19.3%）（P < 0.05）。当线粒体Ca ~（2+）单向转运体的抑制剂钌红（RR）仅存在于IVM培养液中时，存活率和IVM率（分别为54.5%和39.4%）显著高于其他玻璃化冷冻组（分别为25.8-38.4%和14.4- 24.2%）。结果表明，CsA、BAPTA-AM和RR能提高猪GV期玻璃化冷冻卵母细胞的存活率和体外成熟率，而Z-VAD不能提高猪GV期玻璃化冷冻卵母细胞的体外成熟率。(C)2008年爱思唯尔公司All rights reserved.
In this study, we examined the effects of inhibitors of mitochondrial permeability transition (MPT), caspase activity, intracellular Ca2+ chelator and mitochondrial Ca2+ uniporter on survival assessed by morphological observation and in vitro maturation (IVM) of porcine vitrified germinal vesicle (GV) oocytes. When vitrified GV oocytes were matured only present in the IVM medium with an MPT inhibitor, cyclosporin A (CsA), the survival and IVM rates (36.1% and 26.8%, respectively) were significantly higher (P < 0.05) than those in the other vitrified groups (10.3-12.3% and 6.2-10.3%, respectively). However, Z-VAD-fmk (Z-VAD), a caspase inhibitor, did not improve the survival and IVM rates (11.7-21.6% and 8.5-155%, respectively). When BAPTA-AM, an intracellular Ca2+ chelator, was present in the IVM medium, the survival and IVM rates of vitrified GV oocytes (34.5-36.2% and 25.0-26.9%, respectively) were significantly higher (P < 0.05) than those in the absent vitrified groups (17.2-24.2% and 12.9-19.3%, respectively). When ruthenium red (RR), an inhibitor of mitochondrial Ca2+ uniporter, was present only in the IVM medium, the survival and IVM rates (54.5% and 39.4%, respectively) were significantly higher than those in the other vitrified groups (25.8-38.4% and 14.4-24.2%, respectively). Furthermore, blastocysts were successfully produced using porcine vitrified GV oocytes matured in the IVM medium with RR after in vitro fertilization.These results suggested that CsA, BAPTA-AM and RR but not Z-VAD have improved the survival and IVM rates of porcine vitrified GV oocytes. (C) 2008 Elsevier Inc. All rights reserved.