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

Figure 3

Left panel shows a BDNF-pH expressing hippocampal neuron in the presence of 50 mM NH₄Cl. Right panel shows the same cell 80 sec after standard ACSF solution was applied. The graph shows the time course of the fractional change of BDNF-pH intensity (background-subtracted delta F/F₀) from pixels within the color-coded regions of interest (ROIs) shown in the panels above. The change from NH₄Cl-containing ACSF to control buffer quenches BDNF-pH within acidic secretory granules. Time-lapse was performed in an inverted microscope with a Hg-lamp and a cooled CCD camera. Neurons were imaged with a 60x 1.45 NA oil-immersion objective, exposure times ranged from 50–100 ms, and images were taken at 1 frame per second (fps).