Noninvasive visualization of molecular events in the mammalian zygote
Noninvasive visualization of molecular events in the mammalian zygote
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DOI：
10.1002/gene.20158
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发表时间：
2005-10-01
期刊：
影响因子：
1.5
通讯作者：
Ogura, A
中科院分区：
文献类型：
--
作者：
Yamagata, K;Yamazaki, T;Ogura, A
受精后，在哺乳动物受精卵中触发了许多分子事件。由于利用哺乳动物配子和受精卵进行生化研究存在固有的困难，这些过程的分子性质目前尚不清楚。我们已经开发了一种方法来可视化这些事件。在体外，将编码与绿色荧光蛋白融合的蛋白的转录的mRNAs显微注射到卵母细胞或胚胎中，观察到荧光信号。利用这项技术，我们成功地获得了活体小鼠和兔胚胎DNA甲基化状态的图像。此外，还获得了第二次减数分裂和第一次有丝分裂过程中纺锤体和核形成的时间推移图像。重要的是，即使在观察之后，显微注射的胚胎也能发育到正常的后代，这表明该技术相对来说是非侵入性的。因此，我们的方法可能有助于阐明受精和植入前发育的分子方面，并基于实时遗传和表观遗传状态，可能成为在植入前选择“高质量”胚胎的工具。创世纪43：71-79,2005。(C)2005年Wiley-Liss，Inc.
Following fertilization, a number of molecular events are triggered in the mammalian zygote. As biochemical studies using mammalian gametes and zygotes have inherent difficulties, the molecular nature of these processes is currently unclear. We have developed a method to visualize these events. In vitro transcribed mRNAs encoding for proteins fused with green fluorescent protein were microinjected into oocytes or embryos and fluorescence signals were observed. Using this technique we succeeded in obtaining images of the DNA methylation status in living mouse and rabbit embryos. Moreover, time-lapse images were acquired of spindle and nuclear formation during second meiosis and first mitosis. Importantly, the microinjected embryos developed to the normal offspring even after observation, suggesting that the technique is relatively noninvasive. Thus, our method may help elucidate the molecular aspects of fertilization and preimplantation development and, based on the real-time genetic and epigenetic status, could become a tool to select "good quality" embryos before implantation. genesis 43:71-79, 2005. (c) 2005 Wiley-Liss, Inc.