Microinjection Method for Analyzing Zebrafish Early Stage Oocytes.

Microinjection Method for Analyzing Zebrafish Early Stage Oocytes.
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分析斑马鱼早期卵母细胞的微注射方法。

DOI:
10.3389/fcell.2021.753642
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发表时间:
2021
影响因子:
5.5
通讯作者:
Mullins MC
Mullins MC
中科院分区:
生物学2区
文献类型:
--
作者:
Kobayashi M;Jamieson-Lucy A;Mullins MC

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在卵子发生的早期阶段积累并建立卵母细胞极性的母体因子在胚胎发育和生殖细胞形成中起关键作用。然而,脊椎动物卵子发生,特别是卵子发生的早期阶段,由于难以获得这些卵母细胞和缺乏分析方法,没有得到很好的理解。在这里,我们报告的显微注射方法分析斑马鱼早期卵母细胞和一些文物时要知道进行卵母细胞注射或分析卵母细胞。使用这种方法,我们成功地将编码荧光标记蛋白的mRNA注射到早期卵母细胞中,并观察到活卵母细胞中的亚细胞定位。该方法有望为进一步研究卵子发生相关基因的功能奠定基础。
Maternal factors which accumulate and establish oocyte polarity during the early stages of oogenesis play key roles in embryonic development, as well as germ cell formation. However, vertebrate oogenesis, especially early stages of oogenesis, is not well understood due to the difficulty of accessing these oocytes and the lack of analytical methods. Here, we report on a microinjection method for analyzing zebrafish early-stage oocytes and some artifacts to be aware of when performing oocyte injections or analyzing oocytes. Using this method, we successfully injected mRNAs encoding fluorescent-tagged proteins into early-stage oocytes and observed subcellular localization in the live oocytes. This method is expected to advance the functional analysis of genes involved in oogenesis.
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