title: Visualization of peptide secretory vesicles in living nerve cells.
abstract: 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.
authors.affiliation: Neurosciences, University of Toledo College of Medicine, Toledo, OH, USA.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Park
authors.forename: Y Peng
authors.initials: YP
authors.lastname: Loh
mesh_terms: D000818:Animals; D002460:Cell Line; D018274:Electroporation; D006624:Hippocampus / Q000166:cytology; D018613:Microscopy, Confocal; D009474:Neurons / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism*; D051381:Rats; D022142:Secretory Vesicles / Q000378:metabolism*
chemical_list: D009479:Neuropeptides