Neural activity in the corticothalamic network is crucial for sensation, memory, decision, and action. Nevertheless, a systematic characterization of corticothalamic functional connectivity has not been achieved. Here, we developed a high throughput method to systematically map functional connections from the dorsal cortex to the thalamus in awake mice by combing optogenetic inactivation with multi-channel recording. Cortical inactivation resulted in a rapid reduction of thalamic activity, revealing topographically organized corticothalamic excitatory inputs. Cluster analysis showed that groups of neurons within individual thalamic nuclei exhibited distinct dynamics. The effects of inactivation evolved with time and were modulated by behavioral states. Furthermore, we found that a subset of thalamic neurons received convergent inputs from widespread cortical regions. Our results present a framework for collecting, analyzing, and presenting large electrophysiological datasets with region-specific optogenetic perturbations and serve as a foundation for further investigation of information processing in the corticothalamic pathway.

皮层神经网络中的神经活动对于感觉，记忆，决定和行动至关重要。然而，尚未实现皮质丘脑功能连接性的系统表征。在这里，我们开发了一种高通量方法，通过结合光遗传学灭活和多通道记录，系统地映射清醒小鼠中从背皮质到丘脑的功能连接。皮层失活导致丘脑活动迅速减少，揭示了地形组织的皮层丘脑兴奋性输入。聚类分析表明，单个丘脑核内的神经元组表现出不同的动力学。灭活的影响随着时间而发展，并受到行为状态的调节。此外，我们发现，一部分丘脑神经元从广泛的皮层区域接收会聚的输入。我们的结果为收集，分析和呈现具有特定区域光遗传学扰动的大型电生理数据集提供了框架，并为进一步研究皮质丘脑途径中的信息处理奠定了基础。