Rajagopalan Vidyalakshmi, Corpuz Elizabeth O, Hubenschmidt Mark J, Townsend Caroline R, Asai David J, Wilkes David E
Department of Biology, Indiana University-South Bend, South Bend, IN, USA.
Methods Mol Biol. 2009;586:283-99. doi: 10.1007/978-1-60761-376-3_16.
Cilia and eukaryotic flagella are important structures required for the motility of cells, the movement of medium across the surfaces of cells, and the connections between the receptor and synthetic portions of sensory cells. The axoneme forms the cytoskeleton of the cilium comprising several hundreds of proteins that assemble into the 9 + 2 arrangement of outer doublet and central pair microtubules, the inner and outer rows of dynein arms, and many other structures. Tetrahymena thermophila is an excellent model organism for the study of cilia and ciliogenesis. The cell is covered by about 1,000 cilia which are essential for survival. Additionally, the Tetrahymena genome is available and targeted genetic manipulations are straightforward. In this chapter, we describe five protocols that examine properties of cilia: (a) measuring mRNA levels to see the effect of deciliation on gene expression; (b) swimming velocity and linearity; (c) ciliary length and density; (d) phagocytosis that occurs through the ciliated oral apparatus; and (e) depolarization-induced ciliary reversal.
纤毛和真核生物鞭毛是细胞运动、介质跨细胞表面移动以及感觉细胞受体与合成部分之间连接所必需的重要结构。轴丝构成纤毛的细胞骨架，由数百种蛋白质组装成9+2排列的外双联微管和中央对微管、内排和外排动力蛋白臂以及许多其他结构。嗜热四膜虫是研究纤毛和纤毛发生的优秀模式生物。该细胞被约1000根对生存至关重要的纤毛所覆盖。此外，嗜热四膜虫的基因组可用，靶向基因操作也很简单。在本章中，我们描述了五个检测纤毛特性的方案：（a）测量mRNA水平以观察去纤毛对基因表达的影响；（b）游泳速度和线性度；（c）纤毛长度和密度；（d）通过有纤毛的口器发生的吞噬作用；以及（e）去极化诱导的纤毛反转。
