The trial of inner hair cell injury by double patch clamp

双膜片钳损伤内毛细胞的试验

• 批准号: 13671769
• 负责人: SUGASAWA Masashi
• 金额: $ 1.98万
• 依托单位: The University of TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671769/
• 关键词: inner hair cell; vacuolation; resting membrane potential; patch clamp; glutamate; 空胞変性; double-patch; 内耳; Patch clamp; 蝸牛; グルタシン酸; カルシウム

Recently, Patch clamp techniques have been introduced into hair cell electrophysiological researches. By the way, mechanically and enzymatic ally isolation of hair cells from organ of Corti is necessary for patch clamp of hair cells.To purpose of this study is to clarify factors of cell injuries in the isolation process and to prevent those injuries. Finally rebuilding of normal environment of inner hair cell by double patch is a last purpose.Two indexes of the cell injuries became clear, vacuolation and resting membrane potential. The vacuolation at base occurs in almost all cells after isolation process. The cells without vacuolation showed good behavior during experiment. Other factor was resting membrane potential. The cells whose resting membrane potential was below -6OmV also showed good behavior during experiment.By using these indexes, we tried to prevent cell injures by changing isolation processes but all trials were in vain.It is impossible to reconstruct normal condition of inner hair cells by double patch because of rigidity of upper surface of inner hair cells. We could not make a seal on upper surface of inner hair cells.Another methodology is necessary for keeping inner hair cell condition in experiment.

近年来，膜片钳技术被引入到毛细胞电生理研究中。另外，机械和酶法分离Corti器毛细胞是进行毛细胞膜片钳的必要条件，本研究的目的是阐明分离过程中细胞损伤的因素，并提出预防措施。最后，通过双层膜片修复内毛细胞的正常环境，细胞损伤的两个指标空泡化和静息膜电位变得清晰。在分离过程后，几乎所有细胞的基部都发生空泡化。无空泡化的细胞在实验过程中表现出良好的行为。其他因素是静息膜电位。静息膜电位低于-6OmV的细胞在实验过程中也表现出良好的行为，利用这些指标，我们试图通过改变分离方法来防止细胞损伤，但所有的实验都是徒劳的，由于内毛细胞上表面的刚性，双膜片法不可能重建正常的内毛细胞状态。内毛细胞的上表面不能封闭，实验中需要另一种方法来保持内毛细胞的状态。

项目成果

Overexpression of ErbB-2 protein in human middle ear cholesteatomas

ErbB-2蛋白在人中耳胆脂瘤中的过表达

• 发表时间: 2004
• 期刊: Laryngoscope 114(11)
• 作者: Sakamoto T;Kondo K;Yamasoba T et al.
• 通讯作者: Yamasoba T et al.

Actions of Subtype-specific Purinergic ligands on Rat spinal Ganglion Neurons.

亚型特异性嘌呤能配体对大鼠脊髓神经节神经元的作用。

• 发表时间: 2004
• 期刊: Acta Otolaryngology Suppl.552
• 作者: Ken Ito et al.

ラットラセン神経節細胞におけるサブスタンスP受容体の機能的発現

大鼠螺旋神经节细胞P物质受体的功能表达

• 发表时间: 2002
• 期刊: Otology Japan 12
• 作者: 伊藤健