In vivo measurement of afferent activity with axon-specific calcium imaging

In vivo calcium imaging from axons provides direct interrogation of afferent neural activity, informing neural representations that a local circuit receives. Unlike in somata and dendrites, axonal recording of neural activity--both electrically and optically--has been difficult to achieve, thus preventing comprehensive understanding of neuronal circuit function. Here, we developed an active transportation strategy to enrich GCaMP6, a genetically encoded calcium indicator (GECI), uniformly in axons with sufficient brightness, signal-to-noise ratio, and photostability to allow robust, structure-specific imaging of pre-synaptic activity in awake mice. Axon-targeted GCaMP6 (axon-GCaMP6) enables frame-to-frame correlation for motion correction in axons and permits subcellular-resolution recording of axonal activity in previously inaccessible deep brain areas. We used axon-GCaMP6 to record layer-specific local afferents without contamination from somata and intermingled dendrites in the cortex. We expect axon-GCaMP6 will facilitate new applications in investigating afferent signals relayed by genetically defined neuronal populations within and across specific brain regions.