在前期研究中本课题组通过吗琳环寡聚核苷酸显微注射斑马鱼胚胎介导的翻译抑制首次得到了Gfi1和Lmo4表达下调的斑马鱼动物模型，观察到内耳发育异常及毛细胞数量的减少，提示Gfi1和Lmo4在内耳发育中扮演重要角色。本课题拟重点研究转录因子Gfi1和Lmo4在毛细胞分化过程中的作用，寻找其上下游基因，探讨基因相互作用关系，建立相关分子调控网络。将利用由小鼠胚胎听囊细胞形成的内耳前体干细胞系作为体外研究毛细胞分化的细胞模型，在细胞层面进一步验证Gfi1和Lmo4在毛细胞分化过程的作用。寻找调控Gfi1和Lmo4基因翻译的microRNAs，并通过在听囊细胞内microRNA不同水平的表达验证microRNAs对毛细胞分化的影响。本课题选用两种模式动物相结合，在动物水平和细胞水平共同探讨转录因子及microRNAs在毛细胞分化过程中的相关性及分子机制。

In the previous study, our research group established zebrafish Gfi1 and Lmo4 gene knock-down models by morphilino modified antisense oligonucleotide (MO) injection for the first time. The abnormal phenotypes of inner ear and hair cells in early stage of zebrafish development suggested that Gfi1 and Lmo4 played critical roles in the inner ear development. This project intends to focus on the effects of transcription factors Gfi1 and Lmo4 in differentiation of hair cells, and finding their upstream and downstream genes to explore the interaction between genes, eventually establishing molecular regulatory networks. We will use inner ear stem cell lines derived from otocysts of mouse embryos as models to further testify the effects of Gfi1 and Lmo4 in vitro. We will also look for microRNAs which participate in post-transcription regulation of Gfi1 and Lmo4, and verify their effects on hair cell differentiation in cell models. In this project, we will use two model animals to discuss the molecular mechanism of transcription factors and microRNAs in differentiation of hair cells in either the animal level or cell level.