Sato Ken-Ichi, Tokmakov Alexander A
Faculty of Life Sciences, Laboratory of Cell Signaling and Development, Department of Molecular Biosciences, Kyoto Sangyo University, Kyoto, Japan.
Methods Mol Biol. 2019;1920:59-73. doi: 10.1007/978-1-4939-9009-2_5.
Studies on the egg plasma membrane-associated tyrosine kinase Src have shed light on the identity of the molecular machinery that is responsible for gamete interaction and possibly fusion in African clawed frog Xenopus laevis. Here we describe our protocol for identifying and analyzing molecular and cellular machinery that contributes to a variety of biological processes in the course of oogenesis, oocyte maturation, egg fertilization, and early embryogenesis in Xenopus. Our current special interest is to evaluate the hypothesis that the oocyte/egg membrane microdomain (MD)-associated uroplakin III-Src system is responsible for mediating sperm-egg membrane interaction/fusion signal to the oocyte/egg cytoplasm to initiate embryonic and zygotic development in this species. Therefore, this chapter contains a brief introduction to biology of oocytes and eggs in Xenopus and addresses the following questions: (1) What is oocyte/egg MD? (2) Why do we study oocyte/egg MD? (3) How to manipulate oocyte/egg MD? (4) What has been achieved by oocyte/egg MD studies? (5) What are the next steps in oocyte/egg MD studies?
对卵质膜相关酪氨酸激酶Src的研究,揭示了非洲爪蟾(非洲爪蟾)中负责配子相互作用以及可能的融合的分子机制的特性。在此,我们描述了用于鉴定和分析在非洲爪蟾卵子发生、卵母细胞成熟、卵子受精和早期胚胎发育过程中,对多种生物学过程有贡献的分子和细胞机制的实验方案。我们当前特别感兴趣的是评估这样一个假说:卵母细胞/卵子膜微区(MD)相关的uroplakin III-Src系统负责将精卵膜相互作用/融合信号介导至卵母细胞/卵子细胞质,从而启动该物种的胚胎和合子发育。因此,本章简要介绍了非洲爪蟾卵母细胞和卵子的生物学,并探讨以下问题:(1)什么是卵母细胞/卵子MD?(2)我们为什么研究卵母细胞/卵子MD?(3)如何操纵卵母细胞/卵子MD?(4)卵母细胞/卵子MD研究取得了哪些成果?(5)卵母细胞/卵子MD研究的下一步是什么?