摘    要：Gata3在听觉系统发育中具有重要作用。Gata家族转录因子在进化中非常保守。它们的主要特点是能够通过锌指DNA结合域与DNA上的 5’-（A/T）Gata(A/G)-3’序列结合。脊椎动物共鉴定出 6个Gata 转录因子。Gata1-6，在不同的组织中表达，对细胞命运决定、分化、增殖和运动等均有重要作用。哺乳动物耳蜗毛细胞从解剖学和功能上区分，可分为两种类型：内毛细胞（Inner hair cell,IHC）和外毛细胞（Outer hair cell,IHC）8-9。外耳的一般功能是接受外界的声音，内耳的一般功能是感受声音。

基于此，本文对成体后条件性敲除Gata3基因观察其对毛细胞的维持作用相关研究进行了综述。本研究通过成体特异性敲除转录因子Gata3，研究转录因子Gata3毛细胞和支持细胞的形态和功能的影响，主要运用免疫荧光技术来研究内耳中条件性敲除Gata3，观察对毛细胞的结构和形态上的变化。本研究发现P1时期的毛细跟对照组正常小鼠的毛细胞一样，没有什么变化，P5时期也是基本上没有变化，但在P23时期的全耳蜗进行免疫荧光观察时发现，与正常小鼠相比毛细胞的数量减少了，内毛细胞和外毛细胞之间的界限消失了，分不出内毛细胞和外毛细胞。所以我们可以判断Gata3的敲除对毛细胞的数量和形态有明显的影响。

关键词：Gata3；毛细胞；形态；数量

Abstract：Gata3 plays an important role in the development of auditory system. The evolutionarily conserved Gata family transcription factors are characterized by their ability to bind to a consensus DNA sequence 5′-(A/T)GATA(A/G)-3′through their conserved zinc finger DNA-binding domains . Six Gata factors (GATA1-6) have been identified in vertebrates and are expressed in different tissues . Mammalian cochlear hair cellscome in two anatomically and functionally distinct types: the outer and inner hair cells (OHCs and IHCs).The general function of the external ear is to receive external sounds, and the inner ear's general function is to feel the voice.

Based on this, we reviewed the related studies on the conditional Gata3 knockout of adult cells to observe the maintenance of hair cells. In this study, we studied the effects of transcription factor Gata3 on the morphology and function of Gata3 hair cells and supporting cells by adult specific knockout transcription factor. Immunofluorescence technique is used to study the conditional knockout of Gata3 in the inner ear, and to observe the changes in the structure and morphology of hair cells. In this study, we found that the capillaries in P1 period did not change as much as that in normal control mice, and there was no change in P5 stage. However, in the P23 period, the whole cochlea was observed by immunofluorescence, and the number of hair cells decreased compared with normal mice, and the boundary between the inner hair cells and the outer hair cells disappeared. The boundary between inner hair cells and outer hair cells disappeared, and the inner hair cells and outer hair cells could not be separated. So we can judge that knockout of Gata3 has a significant effect on the number and morphology hair cells.

Keyword：Gata3； Hair cells；Form；quantity

目录

1.引言 1

1.1内耳结构和毛细胞 1

1.2关于毛细胞再生的研究 2

1.3 Gata3 3

2.实验的内容与设计 4

2.1 研究的目的 4

2.2研究意义 4

2.3主要研究内容 5

2.4实验技术路线 5

3.实验材料与方法 6

3.1实验动物 6

3.2主要实验试剂 6

3.2.1 小鼠基因型鉴定需要的试剂 6

3.2.2主要试剂配置 7

3.2.3免疫组化试剂 8

附表1.抗体及其工作浓度 9

3.2.4全耳蜗的免疫化（Whole Mount） 9

3.3主要实验设备仪器 9

3.4实验方法 10

3.4.1 小鼠的基因型鉴定： 10

3.4.2实验小鼠材料的准备 11