摘要Lmo4b是一类广泛存在于动物中调控DNA转录的调节蛋白，主要对相关参与感觉器官及大脑神经细胞增殖及发育的基因在时空上进行负调控；先前已有研究人员在小鼠中阐明了哺乳动物Lmo4b 的作用机理，因而在动物模型斑马鱼中研究其在由内耳毛细胞损伤引起的神经性耳聋中的作用对加深该病的了解具有一定意义。我们以DAT转GFP的斑马鱼为研究平台，利用Morphlino转录抑制、Confocal成像及原位杂交等技术对斑马鱼Lmo4b对内耳及脑部的调控作用做初步研究。我们发现Lmo4b会对斑马鱼脑部的大小发育产生影响，特别是前脑，但其调控脑部大小的作用与DAT关系不大；另外，内耳对Lmo4b的调控较为敏感，Lmo4b的抑制与过表达均会影响内耳形态与结构正常发育，对内耳毛细胞的原位杂交也显示，斑马鱼内耳毛细胞的早期发育受到Lmo4b的调控。44277

Abstract Lmo4b is a kind of regulatory proteins that widely exists in animals that regulation DNA transcription,Mainly negative regulation the genes of involving sensory organs development and neuronal proliferation in time and space.Previous studies in the mouse has demonstrated that the mechanism of Lmo4b for mammal,and have a certain significance for sensorinenral deafness that caused by irreversible loss of inner hair cells in the zebrafish model.We take the zebrafish that DAT transferred GFP as a research platform,We discuss the mechanism of Lmo4b act on zebrafish inner and brain,utilize transcription inhibition of Morphlino、Confocal imaging and whole mount in situ hybridization.We propose that Lmo4b impact on the size of zebrafish brain,forebrain particulary.However,the function of regulation the size of brain is independent of DAT. In addition,the zebrafish inner is sensitive to Lmo4b.The Lmo4b knockdown and overexpression may produce an effcet on the morphological structure of zebrafish inner,and the results of inner hair cell in situ hybridization show that the early development of zebrafish inner cell has regulated by Lmo4b.

毕业论文关键字：斑马鱼； 内耳毛细胞； 神经性耳聋； Lmo4b；

Keywords：Zebrafish; Inner hair cells; Sensorineural deafness; Lmo4b

 目次缩略语表 V

1. 引言： 1

1.1 内耳与神经性耳聋 1

1.2动物模型内耳研究的优势 1

1.2.1 小鼠模型 1

1.2.2 斑马鱼模型 1

1.3内耳发育相关基因及分子调控机制 2

1.3.1  ISL1与Pou4f1 2

1.3.2  信号通路 3

1.3.3  Myosin VI 3

1.4内耳发育的负调控. 4

2. 论文研究内容与意义 5

2.1 论文背景与意义 5

2.2．实验内容 5

2.3. 实验流程图 6

3. 仪器设备 6

4. 实验试剂与方法 7

4.1 斑马鱼资源管理 7

4.2 试剂与配方 7

4.3 实验方法