Mitochondrial aggregation caused by cytochalasin B compromises the efficiency and safety of three-parent embryo.
Mitochondrial aggregation caused by cytochalasin B compromises the efficiency and safety of three-parent embryo.
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细胞松弛素 B 引起的线粒体聚集损害三亲胚胎的效率和安全性
DOI：
10.1093/molehr/gaac036
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发表时间：
2022-10-28
影响因子：
4
通讯作者：
Ji, Yazhong
中科院分区：
文献类型：
--
作者：
Li, Ying;Shi, Sanbao;Yuan, Jin;Xiao, Xi;Ji, Dongmei;Pan, Jianxin;Min, Zhunyuan;Wang, Hao;Sha, Hongying;Ji, Yazhong
细胞松弛素B（CB）是卵母细胞去核过程中稳定细胞质所必需的。然而，CB处理导致线粒体分布不均匀，向细胞核聚集，这可能会损害三亲胚胎的效率和安全性。在这里，我们证明了CB处理影响线粒体动力学，纺锤体形态和线粒体DNA携带在浓度依赖性的方式。我们的研究结果表明，小鼠卵母细胞处理超过1 μg/ml CB表现出更多的线粒体聚集模式和减少丝状肌动蛋白的表达。随着CB浓度的升高，线粒体的异常分裂和纺锤体形态的变化也随之增加。在小鼠实验结果的基础上，我们进一步揭示了这些发现在人类卵母细胞中的实用价值。基于芯片的数字PCR和焦磷酸测序显示，通过在纺锤体移植和原核移植前将CB的浓度从标准的5 μg/ml改变为1 μg/ml，重构人胚胎中的线粒体携带显著降低。总之，我们的研究结果为提高线粒体替代疗法的效率和安全性提供了最佳操作。
It is widely accepted that cytochalasin B (CB) is required in enucleation of the oocyte in order to stabilize the cytoplasm. However, CB treatment results in the uneven distribution of mitochondria, with aggregation towards the nucleus, which might compromise the efficiency and safety of a three-parent embryo. Here, we demonstrated that CB treatment affected mitochondrial dynamics, spindle morphology and mitochondrial DNA carryover in a concentration-dependent manner. Our results showed that mouse oocytes treated with over 1 μg/ml CB exhibited a more aggregated pattern of mitochondria and diminished filamentous actin expression. Abnormal fission of mitochondria together with changes in spindle morphology increased as CB concentration escalated. Based on the results of mouse experiments, we further revealed the practical value of these findings in human oocytes. Chip-based digital PCR and pyrosequencing revealed that the mitochondrial carryover in reconstituted human embryos was significantly reduced by modifying the concentration of CB from the standard 5 μg/ml to 1 μg/ml before spindle transfer and pronuclear transfer. In conclusion, our findings provide an optimal manipulation for improving the efficiency and safety of mitochondrial replacement therapy.
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影响因子：
82.9
作者：
Gammage PA;Viscomi C;Simard ML;Costa ASH;Gaude E;Powell CA;Van Haute L;McCann BJ;Rebelo-Guiomar P;Cerutti R;Zhang L;Rebar EJ;Zeviani M;Frezza C;Stewart JB;Minczuk M
通讯作者：
Minczuk M
影响因子：
3.8
作者：
Fulka, H
通讯作者：
Fulka, H
影响因子：
4.6
作者：
Azoury, Jessica;Lee, Karen Wingman;Verlhac, Marie-Helene
通讯作者：
Verlhac, Marie-Helene
影响因子：
13.3
作者：
May-Panloup, Pascale;Boucret, Lisa;Reynier, Pascal
通讯作者：
Reynier, Pascal
影响因子：
11.8
作者：
Deng, Manqi;Suraneni, Praveen;Li, Rong
通讯作者：
Li, Rong