Deciphering the molecular networks that govern the neurogenic potential of astrocytes in the adult mouse brain

成年哺乳动物的大脑含有特殊的星形胶质细胞，作为神经干细胞，能够在整个生命中产生功能性神经元。然而，由于构成性成人神经发生的解剖学限制在少数神经发生区域，大脑只有有限的再生能力。近年来研究发现，成年小鼠纹状体中的星形胶质细胞在脑卒中或Notch信号衰减后体内具有产生新神经元的内在能力。虽然纹状体星形胶质细胞诱导神经发生可能为长期的脑修复提供新的途径，但控制这一过程的分子和细胞机制需要首先探索。在这里，我建议使用创新的分子工具来研究控制纹状体星形胶质细胞命运决定的细胞内和细胞外网络。首先，我将提供一种创新的解决方案，将基于crispr的内源性条形码生成与原位单细胞RNA-seq读出相结合。其次，我将使用这种方法来重建星形细胞谱系，并在解剖学背景下确定潜在的转录网络。第三，基于获得的见解，我将在成年小鼠中使用体内CRISPR基因编辑来操纵纹状体星形胶质细胞命运决定的分子逻辑，以产生欲望的细胞系。我的方法将对控制纹状体星形胶质细胞体内神经发生的分子和细胞网络产生一个全面的观点，并打算为神经元修复策略提供新的途径。

The adult mammalian brain contains specialized astrocytes that act as neural stem cells capable of generating functional neurons throughout life. However, due to the anatomical restriction of constitutive adult neurogenesis to a few neurogenic regions, the brain has only a limited capacity to regenerate. It was recently discovered that astrocytes in the striatum of adult mice possess an intrinsic capacity to generate new neurons in vivo after stroke or attenuation of Notch signaling. While the induction of neurogenesis from striatal astrocytes could provide new avenues for brain repair on the long term, the molecular and cellular mechanisms that govern the process need to be explored first. Here, I propose to use innovative molecular tools to investigate the cell-intrinsic and extracellular networks that govern fate decisions of striatal astrocytes in vivo. First, I will provide an innovative solution to combine CRISPR-based endogenous barcode generation with in situ single cell RNA-seq readout. Second, I will use this approach to reconstruct astrocyte lineages and to identify the underlying transcriptional networks within an anatomical context. Third, based on the obtained insights, I will employ in vivo CRISPR gene editing in adult mice to manipulate the molecular logic underlying fate decisions in striatal astrocytes in order to generate a cell lineage of desire. My approach will yield a comprehensive view on the molecular and cellular networks that govern in vivo neurogenesis from striatal astrocytes and intends to provide new routes for neuronal repair strategies.