Chen Chenyu, Chen Binjun, Qian Xiaoqing, Sun Haojie, Fu Xiao, Ren Dongdong
ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, China.
NHC Key Laboratory of Hearing Medicine Research, Eye & ENT Hospital, Fudan University, Shanghai, China.
Bio Protoc. 2025 Jan 20;15(2):e5167. doi: 10.21769/BioProtoc.5167.
The organ of Corti, located in the inner ear, is the primary organ responsible for animal hearing. Each hair cell has a V-shaped or U-shaped hair bundle composed of actin-filled stereocilia and a kinocilium supported by true transport microtubules. Damage to these structures due to noise exposure, drug toxicity, aging, or environmental factors can lead to hearing loss and other disorders. The challenge when examining auditory organs is their location within the bony labyrinth and their small and fragile nature. This protocol describes the dissection procedure for the cochlear organ, followed by confocal imaging of immunostained endogenous and fluorescent proteins. This approach can be used to understand hair cell physiology and the molecular mechanisms required for normal hearing. Key features • Protocol for the microdissection of the organ of Corti and suitable preparation for later immunostaining. • This technique involves the evaluation of mouse cochlea for planar-cell-polarity protein. • Quantitative and qualitative analysis of hair cell cilia in different dimensions.
位于内耳的柯蒂氏器是动物听觉的主要负责器官。每个毛细胞都有一个由充满肌动蛋白的静纤毛和由真正的运输微管支撑的动纤毛组成的V形或U形毛束。由于噪声暴露、药物毒性、衰老或环境因素对这些结构造成的损伤会导致听力损失和其他疾病。检查听觉器官时面临的挑战在于它们位于骨迷路内，且体积小、质地脆弱。本方案描述了耳蜗器官的解剖程序，随后是对内源性和荧光蛋白进行免疫染色后的共聚焦成像。这种方法可用于了解毛细胞生理学以及正常听力所需的分子机制。关键特性 • 柯蒂氏器显微解剖及后续免疫染色合适制备的方案。 • 该技术涉及对小鼠耳蜗平面细胞极性蛋白的评估。 • 对不同维度的毛细胞纤毛进行定量和定性分析。
