Varjabedian Ani, Kita Angela, Bement William
Laboratory of Cell and Molecular Biology, University of Wisconsin-Madison, Madison, WI, United States; Graduate Program in Cell and Molecular Biology, University of Wisconsin-Madison, Madison, WI, United States.
Graduate Program in Cell and Molecular Biology, University of Wisconsin-Madison, Madison, WI, United States; Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI, United States.
Methods Cell Biol. 2018;144:259-285. doi: 10.1016/bs.mcb.2018.03.013. Epub 2018 Apr 25.
Xenopus laevis has long been a popular model for studies of development and, based on the use of cell-free extracts derived from its eggs, as a model for reconstitution of cell cycle regulation and other basic cellular processes. However, work over the last several years has shown that intact Xenopus eggs and embryos are also powerful models for visualization and characterization of cell cycle-regulated cytoskeletal dynamics. These findings were something of a surprise, given that the relatively low opacity of Xenopus eggs and embryos was assumed to make them poor subjects for live-cell imaging. In fact, however, the high tolerance for light exposure, the development of new imaging approaches, new probes for cytoskeletal components and cytoskeletal regulators, and the ease of microinjection make the Xenopus oocytes, eggs, and embryos one of the most useful live-cell imaging models among the vertebrates. In this review, we describe the basics of using X. laevis as a model organism for studying cell division and outline experimental approaches for imaging cytoskeletal components in vivo in X. laevis embryos and eggs.
非洲爪蟾长期以来一直是发育研究的常用模型，并且基于其卵源无细胞提取物的应用，它也是细胞周期调控及其他基本细胞过程重建研究的模型。然而，过去几年的研究表明，完整的非洲爪蟾卵和胚胎也是用于可视化和表征细胞周期调控的细胞骨架动力学的强大模型。鉴于非洲爪蟾卵和胚胎相对较低的透明度被认为使其成为活细胞成像的不良对象，这些发现有些出人意料。然而，事实上，其对光暴露的高耐受性、新成像方法的发展、用于细胞骨架成分和细胞骨架调节剂的新型探针以及显微注射的简便性，使得非洲爪蟾卵母细胞、卵和胚胎成为脊椎动物中最有用的活细胞成像模型之一。在本综述中，我们描述了将非洲爪蟾用作研究细胞分裂的模式生物的基础知识，并概述了在非洲爪蟾胚胎和卵中对细胞骨架成分进行体内成像的实验方法。
