Visualization of Peptide Secretory Vesicles in Living Nerve Cells
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Analysis of real-time movements of peptidergic vesicles in live neurons provides insight into molecular mechanism(s) supporting
the activity-dependent secretion of neurotrophins and neuropeptides. We examined the effect of overexpression of exogenous
peptides comprising of the cytoplasmic tail sequence of vesicular carboxypeptidase E (CPE), proposed to be involved in the
mechanism of trafficking of peptidergic secretory vesicles, in live hippocampal neurons. E16 rat hippocampal neurons were
transfected with the peptidergic vesicle markers, CPE C-terminally tagged with red or green fluorescent protein, or brain-derived
neurotrophic factor (BDNF) tagged with green fluorescent protein, and grown on dishes specialized for real-time live cell
visualization. Movements of peptidergic vesicles were imaged in a temperature-controlled chamber on a confocal inverted microscope
and analyzed with respect to their velocity, displacement distance, and processivity.
