Multiple Approaches to Investigate the Transport and Activity-Dependent Release of BDNF and Their Application in Neurogenetic Disorders
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  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  for details. Modified from Figure 4(e) in .
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