Multiple Approaches to Investigate the Transport and Activity-Dependent Release of BDNF and Their Application in Neurogenetic Disorders

Figure 2

Schematic illustration of how BDNF-pHluorin fluoresces, both inside the vesicle and upon axonal and dendritic vesicular fusion. Before stimulation, BDNF-pHluorin protein shows little fluorescence. If intracellular calcium increase occurs upon electrical stimulation, then BDNF-pHluorin vesicle may fuse, opening up to the pH 7.4 extracellular space, causing a transient spike in fluorescence that can be detected by TIRF microscopy. Different styles of fusion between axon and dendrite are shown, as explained in [10] and in the text. After sustained vesicular fusion as occurs in dendrites, fluorescence will decrease as a result of BDNF-pHluorin diffusion out of vesicles. Kiss-and-run fusion as occurs in stimulated axons will rather show an increase in fluorescence because minimal pHluorin diffuses out of the vesicle. The sticks represent synaptotagmin-4; see [85] for details. Modified from Figure 4(e) in [85].