脑梗死后神经元损伤和死亡导致功能缺失，如何运用神经元替代和再生进行修复是目前研究的热点。谱系重编程是直接将成熟细胞转分化为其他类型的功能细胞。本课题组前期研究发现，小鼠脑梗死后皮层病灶周围和丘脑远隔部位大量增生的应激性星形胶质细胞表现出神经干细胞的特性，但能否通过重编程将梗死后应激性星形胶质细胞诱导为功能性神经元目前尚不清楚。本项目拟采用脑梗死小鼠模型，应用调控神经胶质前体细胞分化的两个关键因子Pax6和Ngn2，将应激性星形胶质细胞重编程为神经元，经细胞示踪技术动态观察细胞转化过程及其是否表达成熟神经元表面标记分子；通过膜片钳检测这些神经元是否整合入局部神经网络并具备活性功能，进而探讨其可能的分子机制，为脑梗死后细胞替代和再生治疗提供实验依据。

Ischemic infarction causes neuronal deficits. Thus, close attention has been paid to neuronal replacement and regeneration after ischemic infarction. Lineage reprogramming, which can be defined as direct conversion of specific differentiated cell type into another lineage, has been a promising approach for producing functional cells. In our previous study, we found out that reactive glial cells induced by ischemic infarction retained the ability to resume proliferation and acquired stem cell properties both in the peri-infarction area and the remote region. However, whether reactive glial cells can be reprogrammed into functional neurons in the environment of ischemic infarction has not been reported. Therefore, we plan to apply a mouse model of ischemic infarction, and use direct reprogramming of reactive glial cells into neurons by force expression of Pax6 and Ngn2, which are two intrinsic neurogenic determinants of astrocyte precursor cells. The identity of reactive glial cells following retroviral transduction will be determined by time-lapse video microscopy. Immunofluescence staining will be performed to track whether reprogrammed neurons express neuronal markers. To confirm the neuronal identity of the reprogrammed neurons, patch clamp recordings will be performed. By investigating the underlying molecular mechanisms, our study may provide an experimental basis for a promising new approach to initiate brain repair cell replacement and regeneration after ischemic infarction.