Singh P, Das A, Li R
Johns Hopkins University, Baltimore, MD, United States.
Methods Cell Biol. 2017;139:23-50. doi: 10.1016/bs.mcb.2016.12.001. Epub 2017 Jan 23.
Diverse cellular functions require the establishment of cell polarity. Although the morphological manifestation of cell polarity can be complex, common principles and conserved regulatory pathways can be studied leveraging the powerful genetic tools in the budding yeast Saccharomyces cerevisiae. A vast array of genetic and cell biological tools and interdisciplinary approaches have been employed to understand the establishment and maintenance of cell polarity in budding yeast. These tools enabled the identification of key molecular components, their mechanisms of action, and the pathway governing the spatiotemporal properties of cell polarity. In this review, we describe a set of useful methods with special focus on the combination of genetic manipulation and quantitative live-cell imaging, as well as advanced microscopy-based quantitative analysis, for studying symmetry-breaking process in budding yeast. Specific examples are included to illustrate the utilities of each method.
多种细胞功能需要建立细胞极性。尽管细胞极性的形态表现可能很复杂,但利用芽殖酵母酿酒酵母中强大的遗传工具,可以研究其共同原理和保守的调控途径。人们已经采用了大量的遗传和细胞生物学工具以及跨学科方法来理解芽殖酵母中细胞极性的建立和维持。这些工具使得关键分子成分、它们的作用机制以及控制细胞极性时空特性的途径得以确定。在这篇综述中,我们描述了一组有用的方法,特别关注遗传操作与定量活细胞成像的结合,以及基于先进显微镜的定量分析,用于研究芽殖酵母中的对称性破缺过程。文中包含了具体例子来说明每种方法的实用性。