Tracking the dynamics of single fused synaptic vesicle proteins from a single ribbon active zone in zebrafish retinal...

Clearance of fused synaptic vesicle components and availability of release sites are important determinants of recovery from short-term synaptic depression. However, the dynamics of release site clearance are not well established. This protocol illustrates single-molecule imaging of an exocytosis reporter, synaptophysin-pHluorin fusion protein (SypHy), by combining two-color laser scanning confocal microscopy with whole-cell patch-clamp recording of retinal bipolar cells from transgenic zebrafish that weakly express SypHy to track the dynamics of newly fused vesicle proteins at the active zone. For complete details on the use and execution of this profile, please refer to Vaithianathan et al. (2019).