IDENTIFICATION AND CLARIFICATION OF THE UNDERLYING MECHANISM OF NOVEL DEANESS GENE BY MUTAGENESIS PROGRAM

诱变程序鉴定和阐明新型Deanness基因的潜在机制

基本信息

批准号：
14370537
负责人：
IKEDA Katsuhisa
金额：
$ 7.04万
依托单位：
JUNTENDO UNIVERSITY (2003)Tohoku University (2002)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

In recent 5 years, twenty novel genes responsible for hereditary deafness have been discovered in foreign countries, but no new identification of deafness gene has never appeared in Japan. It will be explained by small size of Japanese families, difficulty in classification of phenotype of deafness, difficulty in standard genetic approach to identify novel genes such as positioning cloning and candidate gene approach. Therefore, we use random mutagenesis project, which includes mutagenesis of genes in mouse and screening of deafness, and identification of the responsible gene in order to discover novel deafness genes. Auditory analysis of the mouse model for deafness provides the underlying mechanism of deafness. Furthermore, this animal research can be applied to human study, leading to possibilities of clinical application.We created point mutation of deafness gene in embryonic stem cell by EUN and screened the mice created by auditory brainstem response. Totally 56 mice of 13 lines could be detected deafness. The detailed analysis of auditory responses by tone burst resulted in 12 animals of slight hearing loss, 22 animals of moderate hearing loss, and 32 animals of severe hearing loss. Based upon histopathological analysis, pathological findings were observed in 13 animal in the organ of Corti, 9 in spiral ganglion cells, 5 in stria vascularis, 4 spiral ligament and 2 in tectorial membrane, and no abnormal finding was observed in 2 animals. We discovered that the gene encoding Ca-ATPase was responsible for de at least one line of these mouse models.

近5年来，国外已发现20种导致遗传性耳聋的新基因，但日本从未出现过新的耳聋基因鉴定。这可以解释为日本家庭规模小，耳聋表型分类困难，标准遗传方法难以识别新基因，例如定位克隆和候选基因方法。因此，我们采用随机诱变方案，包括小鼠基因诱变和耳聋筛查，并鉴定相关基因，以发现新的耳聋基因。对耳聋小鼠模型的听觉分析提供了耳聋的潜在机制。此外，这项动物研究还可以应用于人体研究，从而带来临床应用的可能性。我们通过EUN在胚胎干细胞中创建了耳聋基因的点突变，并筛选了听觉脑干反应产生的小鼠。 13系共56只小鼠可检出耳聋。通过音调爆发对听觉反应进行详细分析，结果显示 12 只动物有轻度听力损失，22 只动物有中度听力损失，32 只动物有重度听力损失。组织病理学结果显示，13只动物柯蒂氏器、9只螺旋神经节细胞、5只血管纹、4只螺旋韧带、2只盖膜有病理改变，2只动物未见异常。我们发现编码 Ca-ATP 酶的基因负责这些小鼠模型中至少一个品系的产生。