Math1蛋白核转运机制的研究

耳蜗毛细胞发育异常是造成感音神经性耳聋最常见的原因之一。Math1基因在哺乳动物耳蜗的生长发育调控过程是不可缺少的正性调控基因，Math1基因表达异常或Math1蛋白转运异常会导致毛细胞发育异常或缺失。目前在基因水平上对Math1的研究比较多，但对其在蛋白水平与转录调控相关的的具体过程知之甚少。申请人对Math1蛋白细胞定位前期研究表明其在胞浆被合成后以囊泡形式被转运到细胞核中，但是该过程的分子机制尚不清楚。本项目拟采用纳米颗粒载体将Math1质粒转染细胞来确定Math1蛋白在细胞中的准确定位；通过活细胞动态观察及电镜等方法确定Math1蛋白的入核方式；通过对转染细胞施加影响囊泡释放的因素观察处理因素对Math1蛋白转运的影响；并观察同不处理因素对Math1蛋白发挥转录的影响。从而阐明Math1蛋白核定位的转运机制，解释Math1发挥转录作用的转运机制，为开发分子靶向新药物治疗提供新思路。

Cochlear hair cell dysplasia is one of the most common reasons that can cause sensorineural hearing loss. Math1 gene is an indispensable positive regulatory gene in the growth and development process of the mammalian cochlea. Both abnormal expression of Math1 gene and transportation of Math1 protein can cause hair cell developmental abnormalities or missing. There were so many reports about Math1 at the gene level in the past decade; however, little study was carried out at the protein level. The applicant's previous preliminary research about subcellular localization of Math1 protein suggests that Math1 protein is synthesised in the cytoplasm and then be transported into the nucleus in the form of vesicles. However, the molecular mechanism of this process is still unclear. The project intends to adopt nanoparticles containing Math1 plasmid which can express EmGFP_Math1 fusion protein to transfect cells to determine the accurate subcellular localization of Math1 protein in living cells; to determine the nuclear trafficking mechanism of Math1 protein using confocal dynamic observation in living cells and electric microscopy observation in the fixed cells; to observe the impact of the treatment factors on Math1 protein transportation process through different treatment to the transfected cells, which can affect the release of the vesicles; and to observe the affect of different treatment factors on the Math1 protein transcription. The research is aimed to clarify the transportation mechanism of Math1 protein to nucleus, to explain the role of Math1 protein transportation in transcription, and to provide new ideas for developing new molecular targeted drug to treat sensorineural hearing loss.