Vitrification negatively affects the Ca2+-releasing and activation potential of mouse oocytes, but vitrified oocytes are potentially useful for diagnostic purposes
Vitrification negatively affects the Ca2+-releasing and activation potential of mouse oocytes, but vitrified oocytes are potentially useful for diagnostic purposes
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DOI：
10.1016/j.rbmo.2019.09.012
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发表时间：
2020-01-01
影响因子：
4
通讯作者：
Heindryckx, Bjoern
中科院分区：
文献类型：
--
作者：
Bonte, Davina;Thys, Vanessa;Heindryckx, Bjoern
研究问题：玻璃化冷冻在多大程度上影响小鼠卵母细胞的Ca 2+释放和激活潜力，这通常用于确定人类精子的卵母细胞激活潜力？设计图：玻璃化冷冻对小鼠卵母细胞激活后钙离子动态和发育能力的影响进行了评估，并与新鲜小鼠卵母细胞进行了比较。此外，在玻璃化-加温过程中的不同时间点的内质网的钙库含量进行了测定。结果：人精子注射入小鼠卵母细胞后，玻璃化温度升高可显著改变精子内钙离子浓度，但对受精率无显著影响（P> 0.05）。Ca 2+动态响应SrCl 2或离子霉素也改变了卵母细胞玻璃化冷冻。相比之下，SrCl 2暴露后的激活率和囊胚率没有受到影响（P > 0.05），而离子霉素暴露后的激活率在玻璃化冷冻-温热的卵母细胞中显著降低（P < 0.05）;囊胚率没有受到影响（P > 0.05）。冷冻保护剂暴露与内质网Ca 2+库含量的强烈下降有关。卵母细胞迅速恢复在温暖和恢复含钙的媒体;阈值面积曲线下的钙离子动力学获得激活率高于90%determined.Conclusions：玻璃化温热小鼠卵母细胞显示减少钙释放潜力卵母细胞激活后，冷冻保护剂暴露所造成的。采用适当的分类标准，这些卵母细胞可用于诊断患者的卵母细胞激活缺陷。评估钙2+信号机制玻璃化温热的人类卵母细胞是必要的。
Research question: To what extent does vitrification affect the Ca2+-releasing and activation potential of mouse oocytes, which are commonly used to determine the oocyte activation potential of human spermatozoa?Design: The effect of mouse oocyte vitrification on Ca2+ dynamics and developmental competence after oocyte activation was assessed and compared with fresh mouse oocytes. Moreover, the Ca2+ store content of the endoplasmic reticulum was determined at different time points during the vitrification-warming procedure. Finally, the Ca2+ pattern induced by cryoprotectant exposure was determined.Results: After human sperm injection into mouse oocytes, Ca2+ dynamics but not fertilization rates were significantly altered by vitrification warming (P < 0.05). Ca2+ dynamics in response to SrCl2 or ionomycin were also altered by oocyte vitrification. In contrast, activation and blastocyst rates after SrCl2 exposure were not affected (P > 0.05), whereas activation rates after ionomycin exposure were significantly lower in vitrified-warmed oocytes (P < 0.05); blastocyst rates were not affected (P > 0.05). Cryoprotectant exposure was associated with a strong drop in endoplasmic reticulum Ca2+ store content. Oocytes rapidly recovered during warming and recovery in Ca2+-containing media; a threshold area under the curve of Ca2+ dynamics to obtain activation rates above 90% was determined.Conclusions: Vitrified-warmed mouse oocytes display reduced Ca2+-releasing potential upon oocyte activation, caused by cryoprotectant exposure. With adapted classification criteria, these oocytes could be used for diagnosing oocyte activation deficiencies in patients. Evaluating the Ca2+-signalling machinery in vitrified-warmed human oocytes is required.