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