Bcl-2转基因小鼠再生视神经向外侧膝状体的生长导向研究

项目名称： Bcl-2转基因小鼠再生视神经向外侧膝状体的生长导向研究

项目编号： No.30872835

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 杨柳

作者单位： 北京大学

项目金额： 30万元

中文摘要： 视神经再生后轴突在脑内的正常投射是一个非常复杂的过程，应该有很多导向分子的参与，本研究的目的就是初步探讨Slit/Robo对视神经轴突生长导向中的作用。研究主要探索Slit/Robo在神经轴突损伤后的表达及可能的导向作用。 研究首先在成年野生型小鼠视神经损伤模型上，分析视神经的轴突和胶质细胞的改变，并通过荧光染色法标记Slit-1、Robo-2在视神经、视交叉、视束的表达，以期发现其损伤后表达的改变，从而进一步研究其导向作用。结果发现，损伤后视神经、视交叉、双侧视束均未见阳性表达。Q-PCR示损伤视神经的Robo-2表达量早期降低，随后呈增高趋势。由于Slit/Robo途径并未如项目预期，我们随后开始探索了另一种导向因子Ephrin b2及其受体EphB1在视神经损伤中的作用。我们已完成Bcl-2小鼠品系的维持，建立了Bcl-2转基因小鼠视神经损伤模型，并探讨导向分子在转基因小鼠中的表达。由于视神经再生导向的研究属于探索性，这方面仍需深入艰难的研究。 本研究增加了视网膜疾病神经损害和保护的研究内容，利用米诺环素等进行视网膜脱离和视网膜色素变性的光感受器细胞凋亡的研究。

中文关键词： 视神经；再生；小鼠；Slit/Robo；Eph-B1

英文摘要： It is a very complicated for the regenerated optic nerve axon to get to the target in the brain. There should be lots of guidance molecules involved. The purpose of this study focused on the primary explore of Slit/Robo expression in optice nerve injury and the axon guidance. We have developed optic nerve trauma models in adult wildtype mice, analyze the histomorphology on optic nerve and glial cells. Using immunofluorecence to identify the Slit-1 and Robo-2 expression on optic nerve, optic chaism and optic tract. The result showed that there's no positive cell in Slit-1、Robo-2 fluorescence staining of optic nerve、chiasm or optic tract after crush. RT-PCR shows that Robo-2 expression decrease in trauma optic nerve, and then increase slightly as time goes by. Since this study is based on the hypothesis that Slit/Robo is a key guidance molecule. The result showed there was not obvious changes after the optic nerv crush. We adjusted the study to the roles for another neuron guidance,that is Ephrin b2 and its receptor EphB1. We have succesfully keep the line of Bcl-2 trangenic mice and established the optic nerve regenration model. As we all know, optic nerve regeneration is very difficult and our study is the primary explore on the guidance of regenerated axon, there's a lot of work to be continued. We shall make it up in our later work. Besides regeneration project, we have performed a series of preliminary studies that protect retina and optic nerve from diseases. We use minocycline in retinal detachment and retinitis pigmentosa mice to protect the photoreceptor cell apoptosis. Minocycline can improve bcl-2 transcription and expression in RP mice retina, and slow down the disease, which is perhaps the mail mechanism of neuron protection.

英文关键词： optic nerve；regeneration；mice；Slit/Robo；Eph-B1