Visualization of mesoscopic brain dynamics using novel fluorescence probes

为了揭示神经元如何在大脑神经网络中相互交流，有必要开发成像方法来选择性地可视化感兴趣的细胞群中的细胞活动。在这里，我通过开发利用基因工程技术在靶细胞中功能化的小分子荧光探针，构建了两种这样的细胞类型选择性成像方法。(i)可视化细胞选择性钙动力学的方法。我已经开发了由报告酶功能化的钙离子成像探针，并使用最佳探针，成功地以细胞选择性的方式可视化钙动态。（ii）可视化突触传递动力学的方法。我已经开发了酸性- ph活化探针，它可以通过使用HaloTag标记技术与感兴趣的神经蛋白结合来监测突触囊泡释放，并且使用最佳探针，成功地可视化了神经活动。

To reveal how neurons communicate with each other in neural networks of the brain, it is essential to develop imaging methods for selectively visualizing cellular activities in cell groups of interest. Here, I have constructed two such cell-type selective imaging methods by developing small-molecular fluorescence probes which are functionalized in target cells using genetic engineering techniques. (i) Method to visualize cell-selective calcium dynamics. I have developed calcium ion imaging probes which are functionalized by reporter enzyme and, using the optimal probe, have successfully visualized calcium dynamics in a cell-selective manner. (ii) Method to visualize synaptic transmission dynamics. I have developed acidic-pH-activatable probes which can monitor synaptic vesicle release by being conjugated to neural protein of interest using HaloTag labeling technology and, with the optimal probe, have successfully visualized neural activities.