Using Xenopus oocyte extracts to study signal transduction.
Using Xenopus oocyte extracts to study signal transduction.
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DOI：
10.1007/978-1-59745-000-3_31
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发表时间：
2006
影响因子：
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通讯作者：
Richard Crane;J. Ruderman
中科院分区：
文献类型：
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作者：
Richard Crane;J. Ruderman
在减数分裂的I前期，异卵母细胞自然停留在G2/M。黄体酮刺激启动非转录信号通路，最终激活Cdc2/细胞周期蛋白B并重新进入减数分裂。该途径为类固醇激素的非基因组信号传导和G2/M细胞周期转变提供了范例。它已广泛研究使用完整的卵母细胞，这是适用于显微注射和生化分析，在本书的其他地方描述。然而，在使用由前驱卵母细胞的细胞质部分组成的体外系统中有几个实验优势。由于其同质性，提取物避免了单个卵母细胞之间信号不同步的困难。它们也适用于生化操作，如蛋白质免疫消耗，蛋白质和药理学试剂可以很容易地添加。尽管有这些特点，卵母细胞提取物还没有达到xenopusegg提取物的广泛应用，它可以在体外进行几轮脱氧核糖核酸（DNA）复制和有丝分裂。在这里，我们回顾了卵母细胞提取物的历史发展，并讨论了在体外成功复制信号通路和G2/M转化的最关键因素。
Xenopusoocytes are naturally arrested at G2/M in prophase I of meiosis. Stimulation with progesterone initiates a nontranscriptional signaling pathway that culminates in the activation of Cdc2/cyclin B and reentry into meiosis. This pathway presents a paradigm for nongenomic signaling by steroid hormones and for the G2/M cell cycle transition. It has been extensively studied using intact oocytes, which are amenable to microinjection and biochemical analyses described elsewhere in this book. However, there are several experimental advantages in using in vitro systems consisting of cytosolic fractions of prophase-arrested oocytes. Because of their homogeneous nature, extracts avoid the difficulties of signaling asynchrony between individual oocytes. They are also amenable to biochemical manipulations such as protein immunodepletions, and proteins and pharmacological agents can be added easily. Despite these features, oocyte extracts have yet to achieve the widespread utility ofXenopusegg extracts, which can proceed through rounds of deoxyribonucleic acid (DNA) replication and mitosis in vitro. Here, we review the historical development of oocyte extracts and discuss the factors most crucial to success in reproducing the signaling pathway and the G2/M transition in vitro.