Trapani Josef G, Nicolson Teresa
Howard Hughes Medical Institute, Oregon Hearing Research Center and Vollum Institute, Oregon Health and Science University, Portland, Oregon, USA.
Methods Cell Biol. 2010;100:219-31. doi: 10.1016/B978-0-12-384892-5.00008-6.
Sensory signal transduction, the process by which the features of external stimuli are encoded into action potentials, is a complex process that is not fully understood. In fish and amphibia, the lateral-line organ detects water movement and vibration and is critical for schooling behavior and the detection of predators and prey. The lateral-line system in zebrafish serves as an ideal platform to examine encoding of stimuli by sensory hair cells. Here, we describe methods for recording hair-cell microphonics and activity of afferent neurons using intact zebrafish larvae. The recordings are performed by immobilizing and mounting larvae for optimal stimulation of lateral-line hair cells. Hair cells are stimulated with a pressure-controlled water jet and a recording electrode is positioned next to the site of mechanotransduction in order to record microphonics--extracellular voltage changes due to currents through hair-cell mechanotransduction channels. Another readout of the hair-cell activity is obtained by recording action currents from single afferent neurons in response to water-jet stimulation of innervated hair cells. When combined, these techniques make it possible to probe the function of the lateral-line sensory system in an intact zebrafish using controlled, repeatable, physiological stimuli.
感觉信号转导是一个将外部刺激特征编码为动作电位的过程，这是一个尚未被完全理解的复杂过程。在鱼类和两栖动物中，侧线器官可检测水的流动和振动，对群体行为以及捕食者和猎物的探测至关重要。斑马鱼的侧线系统是研究感觉毛细胞对刺激进行编码的理想平台。在此，我们描述了使用完整的斑马鱼幼体记录毛细胞微音器电位和传入神经元活动的方法。记录是通过固定和安装幼体以实现对侧线毛细胞的最佳刺激来进行的。用压力控制的水射流刺激毛细胞，并将记录电极放置在机械转导部位旁边，以记录微音器电位——由于通过毛细胞机械转导通道的电流而产生的细胞外电压变化。通过记录单个传入神经元对受支配毛细胞的水射流刺激产生的动作电流，可获得毛细胞活动的另一种读数。将这些技术结合起来，就能够使用可控、可重复的生理刺激来探究完整斑马鱼中侧线感觉系统的功能。
