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STUDY ON HAIR CELL REGENERATION IN THE MAMMALIAN COCHLEA

哺乳动物耳蜗毛细胞再生的研究

基本信息

批准号：
13470357
负责人：
YAMASOBA Tatsuya
金额：
$ 6.4万
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 (B)
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/en/grant/KAKENHI-PROJECT-13470357/
关键词：
HAIR CELL SUPPORTING CELL COCHLEA ORGAN OF CORTI MITOSIS REGENERATION GUINEA PIG Math1 カスパーゼ

项目摘要

1)We examined the types of cells that can be tranfected with adenovirus vector. We injected an Adex1CAlacZ into the guinea pig cochlear or vestibular explants. We found expression of lacZ in a variety of cochlear tissues including the spiral ganglion, stria vascularis, and the outer hair cells and supporting cells. In the vestibular labyrinth, the mesothelial cells in the perilymphatic space, the sensory and supporting cells in the utricle and ampulla and the transitional and dark cells in the ampulla were transfected. In vivo, we found that damage from a local anesthetic solution containing phenol permitted LacZ gene delivery to the guinea pig inner ear via the round window membrane.2)We investigated the effects of dexamethasone-induced immunosuppression on the outcome of adenovirus gene transfer in guinea pig inner ears and found that immunosuppressive treatment reduced the negative consequences of repeated adenovirus-mediated gene therapy.3) We demonstrate that in vivo inoculation of adenovirus with the Math 1 gene insert into the mature guinea pig cochlea results in Math 1 overexpression in nonsensory cochlear cells. Math 1 overexpression leads to the appearance of immature hair cells in the organ of Corti and new hair cells adjacent to the organ of Corti in the interdental cell, inner sulcus, and Hensen cell regions. Axons are extended from the bundle of auditory nerve toward some of the new hair cells, suggesting that the new cells attract auditory neurons.4)We assessed mitotic activity in the auditory sensory epithelium throughout the mature cochlea of deafened guinea pigs given BrdU for up to 10 days. We found BrdU-positive cells within the organ of Corti from 3 to 30 days following deafening. Immunohistochemistry confirmed that the BrdU-positive cells were the Deiters cells, These findings indicate that, albeit limited, the Deiters cells in the mature mammalian cochlea possess a potential of mitotic activity and that they could survive after mitosis.

1）我们检查了可以用腺病毒载体转染的细胞类型。我们将Adex1CAlacZ注入豚鼠耳蜗或前庭外植体。我们发现lacZ的表达在各种耳蜗组织，包括螺旋神经节，血管纹，外毛细胞和支持细胞。在前庭迷路中，转染外淋巴间隙中的间皮细胞、椭圆囊和壶腹中的感觉细胞和支持细胞以及壶腹中的移行细胞和暗细胞。在体内，我们发现，含有苯酚的局部麻醉溶液造成的损伤允许LacZ基因通过圆窗膜传递到豚鼠内耳。2）我们研究了地塞米松诱导的免疫抑制对豚鼠内耳中腺病毒基因转移结果的影响，发现免疫抑制治疗减少了重复腺病毒介导的基因治疗的负面后果。3）我们证明，在体内接种的腺病毒与数学1基因插入到成熟的豚鼠耳蜗的结果数学1过表达的非感觉耳蜗细胞。Math 1过表达导致Corti器官中未成熟毛细胞的出现以及齿间细胞、内沟和Hensen细胞区域中邻近Corti器官的新毛细胞的出现。轴突从听神经束延伸到一些新的毛细胞，这表明新的细胞吸引听觉神经元。4）我们评估了给予BrdU长达10天的麻醉豚鼠的整个成熟耳蜗的听觉感觉上皮细胞中的有丝分裂活性。我们发现，在Corti器官内的BrdU阳性细胞从3到30天后，震耳欲聋。免疫组化证实BrdU阳性细胞为Deiters细胞。这些结果表明，成熟哺乳动物耳蜗中的Deiters细胞具有有丝分裂活性，并且在有丝分裂后仍能存活。