The research for inner ear specific proteins and deafness gene coding proteins.

内耳特异性蛋白及耳聋基因编码蛋白的研究。

基本信息

批准号：
16591702
负责人：
TAKUMI Yutaka
金额：
$ 2.3万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591702/
关键词：
inner ear deafness gene COL9A3 CRYM thyroid hormone Na, K-ATPase β1 Ubiquitin A-52 K^+ recycling UbA52遺伝子難聴蝸牛血管条辺縁細胞前庭暗細胞 Na,K-ATPase 甲状腺ホルモン IX型コラーゲン蛋白ノックアウトマウス

项目摘要

Through cDNA microarray analysis of gene expression in the cochlea and vestibule, a number of genes that are highly expressed specifically in auditory tissues have been detected (Abe et al., 2003). Moreover, screening for genetic alterations in these genes has identified several possible disease-causing mutations. Here we targeted the COL9A3 (type IX collagen), mu-crystallin (CRYM) and ubiquitin A-52 residue ribosomal protein fusion product 1 (UbA52) genes that are highly expressed specifically in the cochlea and vestibule. The localization of the proteins coded by these genes and the mechanism of hearing disorders were examined in the inner ear.Immunocytochemical analysis revealed that type IX collagen is distributed in the tectorial membrane of the organ of Corti. To confirm the significance of type IX collagen for normal hearing, we assessed the detailed morphological and electrophysiological features of type IX collagen knock-out mice, which have recently been reported as a deafnes … More s model. Through assessment by auditory brainstem response (ABR), knock-out mice were shown to have progressive hearing loss.In a search for mutations of mu-crystallin (CRYM), a taxion specific crystalline which is also known as an NADP regulated thyroid hormone binding protein, two mutations were found at the C-terminus in patients with non-syndromic deafness. T3 binding activity of mutant mu-crystallin was compared with that of wild-type mu-crystallin, and one mutant was shown to have no binding capacity for T3, indicating that CRYM mutations cause auditory dysfunction through thyroid hormone binding properties. Immunocytochemical results indicated that mu-crystallin was co-localized with Na,K-ATPase within type II fibrocytes of the lateral wall. Therefore, CRYM mutations may cause auditory dysfunction through thyroid hormone binding effects on the fibrocytes of the cochlea. mu-Crystallin may be involved in the potassium ion recycling system together with Na,K-ATPase.The ubiquitin A-52 residue ribosomal protein fusion product 1 (UbA52) was found by immunocytochemical investigation to be distributed in the strial marginal cells and vestibular dark cells, which regulate the endolymphatic ion homeostasis. In the developing mouse cochlea, no significant staining was observed from birth to postnatal day 3, whereas after postnatal day 6, strong UbA52-immunoreactivities were observed in strial marginal cells. Endolymphatic K + concentration is elevated between postnatal days 3-8, therefore, our results indicate that UbA52 may have a functional role in regulation of ion secretion in the inner ear. Less

通过对耳蜗中基因表达和可有效的cDNA微阵列分析，已经检测到许多在听觉组织中高度表达的基因（Abe等，2003）。此外，对这些基因的遗传改变的筛查已经确定了几种可能引起疾病的突变。在这里，我们针对的是COL9A3（IX型胶原蛋白），MU-晶状体蛋白（CRY）和泛素A-52残基核糖体蛋白融合产物1（UBA52）基因，这些基因（UBA52）基因在同时有特异性表达并有效。这些基因编码的蛋白质的定位以及在内耳中检查了听力障碍的机理。免疫细胞化学分析表明，IX型胶原蛋白分布在Corti器官的tectorial膜中。为了确认IX型胶原蛋白在正常听力方面的重要性，我们评估了IX型胶原蛋白敲除小鼠的详细形态和电生理特征，这些小鼠最近据报道是聋哑人……更多的S模型。通过听觉脑干反应（ABR）评估，敲除小鼠的听力损失显示。在寻找MU-晶状体蛋白（CRY）突变（CRYS）的突变，这是一种特定纳税的晶体，该突变也被称为NADP调节的甲状腺甲状腺同源型结合蛋白，在非疾病中的C-Terminus中发现了两个突变。将突变体Mu-晶状蛋白的T3结合活性与野生型MU-晶状体蛋白的结合活性进行了比较，并且显示一个突变体的T3没有结合能力，表明CRYM突变会通过甲状腺激素结合特性引起听觉功能障碍。免疫细胞化学结果表明，在侧壁的II型纤维细胞中，MU-晶状体与Na，K-ATPase共定位。因此，CRYM突变可能通过甲状腺激素结合对耳蜗的纤维细胞引起听觉功能障碍。 Mu-晶状蛋白可能与Na，K-ATPase一起参与钾离子回收系统。泛素A-52残基核糖体蛋白融合产物1（UBA52）通过免疫细胞化学研究发现，在细胞块状细胞和前庭暗细胞中分布在调节的nion中，该研究分布在细胞质细胞和前庭暗细胞中。在发育中的小鼠耳蜗中，从生日到产后第3天未观察到明显的染色，而在产后第6天后，在分层的边缘细胞中观察到强大的UBA52-免疫反应性。内淋巴K +浓度在产后3-8天之间升高，因此，我们的结果表明UBA52在内耳内离子分泌的调节中可能具有功能性作用。较少的