A novel technique for inducing local inner ear damage
A novel technique for inducing local inner ear damage
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DOI：
10.1016/s0378-5955(02)00768-2
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发表时间：
2003-02-01
期刊：
影响因子：
2.8
通讯作者：
Ito, J
中科院分区：
文献类型：
--
作者：
Nakagawa, T;Kim, TS;Ito, J
随着小鼠基因组学的显著发展以及转基因和基因敲除小鼠的出现，小鼠将成为内耳研究的首选动物模型。然而，由于小鼠对耳毒性药物的相对抗性和手术治疗的困难，很少有研究将小鼠作为实验动物来检查毛细胞变性。本研究建立了后半规管注射新霉素诱导小鼠内耳感觉上皮细胞凋亡的模型。末端脱氧核苷酸转移酶介导的dUTP镍端标记（TUNEL）实验显示，局部应用新霉素可在一定时间内充分诱导听觉和前庭上皮细胞凋亡。补充一般caspase抑制剂可显著减少tunel阳性细胞的诱导，表明在本模型中观察到的凋亡细胞死亡依赖于caspase。此外，后半规管入路是一种简单的技术，假手术没有引起tunel阳性细胞的明显诱导。因此，该模型能够在毛细胞凋亡机制的研究中使用各种遗传工具。(C) 2002 Elsevier Science B.V.版权所有
With significant development of mouse genomics and the availability of transgenic and knockout mice, the mouse will be the preferred animal model for inner ear research. However, few studies have used mice as experimental animals for examination of hair cell degeneration, because of their relative resistance to ototoxic agents and difficulties in surgical treatment. This study presents a model for induction of apoptotic cell death in sensory epithelia of the mouse inner ear using injection of neomycin into the posterior semicircular canal. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay revealed that local application of neomycin produced sufficient induction of apoptotic cell death in both auditory and vestibular epithelia over a definite time course. Supplementation of the general caspase-inhibitor significantly reduced induction of TUNEL-positive cells, indicating caspase-dependency of apoptotic cell death observed in the present model. In addition, the approach to the posterior semicircular canal was an easy technique, and sham-operation induced no significant induction of TUNEL-positive cells. This model, hence, enables the use of various genetic tools in studies for mechanisms of hair cell apoptosis. (C) 2002 Elsevier Science B.V. All rights reserved.