Wakabayashi Ken-ichi, Kamiya Ritsu
Chemical Resources Laboratory, Tokyo Institute of Technology, Yokohama, Japan.
Department of Life Science, Faculty of Science, Gakushuin University, Tokyo, Japan.
Methods Cell Biol. 2015;127:387-402. doi: 10.1016/bs.mcb.2014.12.002. Epub 2015 Mar 6.
Motile cilia and flagella rapidly propagate bending waves and produce water flow over the cell surface. Their function is important for the physiology and development of various organisms including humans. The movement is based on the sliding between outer doublet microtubules driven by axonemal dyneins, and is regulated by various axonemal components and environmental factors. For studies aiming to elucidate the mechanism of cilia/flagella movement and regulation, Chlamydomonas is an invaluable model organism that offers a variety of mutants. This chapter introduces standard methods for studying Chlamydomonas flagellar motility including analysis of swimming paths, measurements of swimming speed and beat frequency, motility reactivation in demembranated cells (cell models), and observation of microtubule sliding in disintegrating axonemes. Most methods may be easily applied to other organisms with slight modifications of the medium conditions.
能动纤毛和鞭毛能快速传播弯曲波，并在细胞表面产生水流。它们的功能对于包括人类在内的各种生物体的生理和发育都很重要。这种运动基于轴丝动力蛋白驱动的外双联微管之间的滑动，并受到各种轴丝成分和环境因素的调节。对于旨在阐明纤毛/鞭毛运动和调节机制的研究，衣藻是一种非常有价值的模式生物，它提供了多种突变体。本章介绍了研究衣藻鞭毛运动的标准方法，包括游泳路径分析、游泳速度和搏动频率测量、去膜细胞（细胞模型）中的运动再激活以及在解体轴丝中观察微管滑动。大多数方法稍作培养基条件的修改就可轻松应用于其他生物体。
