我们前期研究发现视网膜Müller细胞分化的神经节细胞轴突生长发育不良甚至缺失。本项目中首先体外研究拟采用miR-124上调轴突导向因子促使分化的神经节细胞轴突的生长发育，并确定其调控机制。其次将体外转染miR-124过表达慢病毒载体的Müller细胞源性干细胞移植到青光眼模型鼠眼内，ERG和VEP检测视功能变化，顺行示踪剂神经纤维3D图像分析分化的神经节细胞轴突生长的长度、形态、迁移导向及整合能力，ISH检测miR-124的分布及表达变化，免疫荧光染色、qPCR和Western-blot等方法检测CoREST、REST、Sema3A、NRP-1 mRNA及蛋白的表达差异。最后采用TUNEL技术分析视网膜细胞的凋亡情况，从而揭示miR-124调控Müller细胞分化的神经节细胞轴突导向生长的分子机制及其副作用，为临床基因治疗青光眼奠定理论和实践基础。

In our previous study, we found that ganglion cells derived from retinal Müller cells have defects in axon growth. In this project we speculate that miR-124 promotes the navigation and growth of retinal ganglion cells derived from Müller cells. To test our hypothesis, we will use miR-124 to induce axon growth of differentiated ganglion cells in vitro, and elucidate the underlying regulatory mechanisms. Next, we will transplant stem cells derived from Müller cells transfected with lentivirus vectors overexpressing miR-124 into the eyes of the glaucoma model rats, ERG and VEP are performed to detect the changes in vivual function, anterograde tracer is used to display 3D image of optic nerve fibers to analyze the ganglion cell axon growth, morphology, migration and the ability to integrate, ISH is performed to detect the miR-124 ’ expression and distribution, Immunofluorescence, qPCR and Western-blot analysis are performed to detect CoREST, REST, Sema3A, NRP-1 mRNA and protein. Finally, TUNEL technique is performed to analyze the apoptosis of retinal cells. The completion of this project will help reveal molecular mechanisms and evaluate side effects of miR-124 in the regulation of axon navigation and growth of ganglion cells derived from Müller cells, and promote translational research of miR-124 in gene therapy of glaucoma.