2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38790In the developed countries, the incidence of congenital hearing loss is estimated at 1 in 1000 births, of which approximately 60% cases are attributed to genetic factors. Previously, we have found many novel mutations in the Cx26 and Cx29 from screening of 262 patients with nonsyndromic deafness. However, functional alterations regarding hearing loss in the mutations of those genes remain unknown. The overall goal of this project is to study the pathogenesis of Cx26 and Cx29 from non-syndromic deafness in Taiwan. Simultaneously, we will also create a zebrafish model to investigate the mechanisms of hearing loss by which the Cx26 and Cx29 mutations are exerted. In the proposed three-year project, by using approaches of molecular biological, cell biological and electron microscopy we will focus on study the functional consequence of Cx26 and Cx29 mutations and functional interaction between them in cell-based system. In addition, we will use the zebrafish model to study the mechanisms of hearing loss by the Cx26 and Cx29 mutations with approaches of bioinformatics, molecular biological, fluorescein-phalloidin stain, DASPEl live stain, whole-mount in situ hybridization of embryo, tissue in situ hybridization, gene knockdown and transgene. Upon completion of this research, we anticipate that the study will further our understanding of the functional role of these genes in nonsyndromic deafness. In addition, we believe that the use of the zebrafish model will provide a good approach to study the etiology and mechanism of pathogenesis in hereditary hearing loss.約有千分之一嬰兒在出生時罹患重度聽障，在已開發國家約有60%個案是遺傳因素。最近針對262位非症候群聽障患者基因篩檢研究中，我們發現在CONNEXIN26(Cx26)和CONNEXIN29(Cx29)基因上有新的突變發生。然而目前對於這些基因突變所產生的突變蛋白對功能所造成的影響和其致病機制並不清楚，且同一基因在不同位置的突變常會產生蛋白功能影響上的差異。因此本計畫的研究動機之一是利用已建立好的細胞模式探討Cx26和Cx29基因這些新突變對其功能的影響和彼此之間所造成的影響。同時利用動物模式來探討人類及動物疾病，是生物醫學的一個重要方向。因此本計畫另一個動機是建立和利用斑馬魚為模式動物來探討Cx26及Cx29基因突變造成聽障的機制。藉有這兩個模式的建立和探討將可釐清這兩個基因在非症候群聽障中的致病機制。本研究計畫將分三年來探討兩大主題：1.我們將利用分子生物學、細胞生物學、電子顯微鏡等研究法來加以探討在非症候群聽障患者中，Cx26和Cx29基因新發現突變所造成功能影響或此兩基因彼此之間的交互作用。2.建立一個以斑馬魚為模式動物，利用生物資訊學方法尋找斑馬魚基因庫中與Cx26或Cx29基因之同源基因或相同基因，再利用分子生物學、螢光活體染色法、全胚胎原位雜交法、組織原位雜交法、基因knockdown技術和基因轉殖(transgene)技術來探討Cx26和Cx29基因突變造成聽障的機制。相信這三年計畫如能獲得支持將對於Cx26和Cx29基因突變造成聽障的機制有深一層的瞭解，另外斑馬魚模式動物的建立和應用將有助於幫助我們釐清和探討基因突變造成非症候群聽障的機制。基因突變聽障功能研究斑馬魚Cx26Cx29mutationhearing losszebrafishgap junctionProject