Kobayashi Manami, Jamieson-Lucy Allison, Mullins Mary C
Department of Cell and Developmental Biology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, United States.
Front Cell Dev Biol. 2021 Nov 8;9:753642. doi: 10.3389/fcell.2021.753642. eCollection 2021.
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.
在卵子发生早期积累并建立卵母细胞极性的母体因素在胚胎发育以及生殖细胞形成过程中发挥着关键作用。然而，由于获取这些卵母细胞存在困难且缺乏分析方法，脊椎动物的卵子发生，尤其是卵子发生的早期阶段，尚未得到充分了解。在此，我们报告一种用于分析斑马鱼早期卵母细胞的显微注射方法以及在进行卵母细胞注射或分析卵母细胞时需要注意的一些假象。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，并在活卵母细胞中观察到亚细胞定位。预计这种方法将推动对卵子发生相关基因的功能分析。
