组蛋白去乙酰化酶(HDACs)调控斑马鱼侧线器发育及其机制研究

项目名称： 组蛋白去乙酰化酶(HDACs)调控斑马鱼侧线器发育及其机制研究

项目编号： No.81500784

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 何英姿

作者单位： 复旦大学

项目金额： 18万元

中文摘要： 组蛋白去乙酰化酶（HDACs）在细胞增殖、分化、存活及早期胚胎发育和特定器官形成中发挥重要作用。我们前期研究发现采用HDACs抑制剂调控HDACs能够显著抑制HDAC1、HDAC3活性，并影响斑马鱼侧线神经丘毛细胞的早期分化和再生，提示HDAC1、HDAC3参与毛细胞分化和再生过程。本课题我们拟采用斑马鱼模型，从基因水平调控HDAC1、HDAC3的表达，通过形态结构分析、原位杂交、免疫组化等方法研究HDAC1、HDAC3在侧线系统形成，细胞增殖和存活及毛细胞分化中的作用。采用原位杂交技术结合信号通路调控等方法探讨其作用的机制。最后进一步应用染色质免疫共沉淀测序（ChIP-Seq）结合转录组测序技术（RNA-Seq）深入研究HDAC1、HDAC3调控侧线发育的分子机制。本项目的研究将有利于深入理解听觉器官的发育调控机制，为感音神经性耳聋的防治提供理论依据。

中文关键词： 发病机制；表观遗传；斑马鱼；侧线器；毛细胞

英文摘要： Histone deacetylases (HDACs) activity has been shown to be important for modulating many biological processes including cellular development and regeneration in vertebrates. We have previously reported important roles for HDACs in hair cell development and regeneration in zebrafish, and found a significant downregulation of HDAC1 and HDAC3 protein levels in zebrafish treated with the HDACs inhibitors. These findings highlight the importance of HDAC1 and HDAC3 for hair cell differentiation and regeneration in zebrafish lateral line system. Here, using gene interference methods, we try to examine the role of HDAC1 and HDAC3 in the development of zebrafish lateral line system, cell proliferation and survival, as well as the differentiation of hair cells. We performed whole-mount in situ hybridization analysis in combination with drug treatment and transgenic zebrafish to understand the effect of HDAC1 and HDAC3 on signaling pathways. We next explored the mechanisms of gene regulation by HDAC1 and HDAC3 in lateral line primordium using chromatin immunoprecipitation coupled with massive parallel sequencing (ChIP-seq) to identify genomic regions of HDAC1 and HDAC3 enrichment. RNA sequencing (RNA-seq) were performed to indicate the differential expression of genes in lateral line primordium. The project will contribute to a better understanding of the processes of auditory organ development and facilitate prevention and treatment of sensorineural hearing loss.

英文关键词： pathogenesis;epigenetic;zebrafish;lateral line;hair cell