Second Division, Department of Internal Medircine, Hamamatsu University School of Medicine, Hamamatsu, Japan.
Second Division, Department of Internal Medircine, Hamamatsu University School of Medicine, Hamamatsu, Japan.
Methods Cell Biol. 2023;176:159-180. doi: 10.1016/bs.mcb.2022.12.015. Epub 2023 Feb 8.
The airway epithelium contains numerous multiciliated cells. The apical surface of multiciliated cells is covered with cilia that move at 15-25Hz. Ciliary movement is not a simple reciprocal movement and distinctly has forward and reverse movements called effective and recovery strokes, respectively. These "asymmetric" ciliary strokes push away the mucus covering the mucosa of the airway epithelium. Mucus flow created by ciliary stroke is important for capturing and expelling dust, pollen, PM2.5, pathogens, and other particles that enter the airways from outside the body. This mechanism for protecting the airways produced by ciliary movement is called mucociliary function. Defects in ciliary motility lead to impairment of mucociliary function, resulting in recurrent airway infections such as bronchitis and pneumonia, and consequently, bronchiectasis. While the analysis of ciliary beat frequency is relatively easy, the analyses of the amplitude, velocities of strokes, and the asymmetric level require specific techniques and tips. In this chapter, we present methods for the analysis of ciliary movements of a group of cilia on the luminal surface of the trachea ex vivo and individually isolated and ATP-reactivated cilia in vitro. In addition, a method for the analysis of mucociliary function is also presented.
气道上皮含有众多的纤毛细胞。纤毛细胞的顶表面覆盖着纤毛，纤毛以 15-25Hz 的频率运动。纤毛运动不是简单的往复运动，而是具有明显的前进和后退运动，分别称为有效运动和恢复运动。这些“不对称”的纤毛运动将覆盖气道上皮黏膜的黏液推开。纤毛运动产生的黏液流对于捕获和排出从体外进入气道的灰尘、花粉、PM2.5、病原体和其他颗粒非常重要。纤毛运动产生的这种保护气道的机制称为黏液纤毛功能。纤毛运动缺陷导致黏液纤毛功能障碍，导致反复发生气道感染，如支气管炎和肺炎，进而导致支气管扩张。虽然分析纤毛的摆动频率相对容易，但分析摆动幅度、运动速度和不对称程度需要特定的技术和技巧。在这一章中，我们介绍了分析离体气管腔面一组纤毛和体外分离和 ATP 再激活的纤毛的纤毛运动的方法。此外，还介绍了分析黏液纤毛功能的方法。
