Regulation of peptidergic vesicle mobility by secretagogues.
Neuropeptides are released into the extracellular space from large secretory granules. In order to reach their release sites, these granules are translocated on microtubules and thought to interact with filamentous actin as they approach the cell membrane. We have used a green fluorescent protein-tagged neuropeptide prohormone (prepro-orphanin FQ) to visualize vesicle trafficking dynamics in NS20Y cells and cultures of primary hippocampal neurons. We found that the majority of secretory granules were mobile and accumulated at both the tips of neurites as well as other apparently specialized cellular sites. We also used live-cell imaging to test the notion that peptidergic vesicle mobility was regulated by secretagogues. We show that treatment with forskolin appeared to increase vesicle rates of speed, while depolarization with high K+ had no effect, even though both treatments stimulated neuropeptide secretion. In cultured hippocampal neurons the green fluorescent protein-tagged secretory vesicles were routed to both dendrites and axons, indicating that peptidergic vesicle transport was not polarized. Basal peptidergic vesicle mobility rates in hippocampal neurons were the same as those in NS20Y cells. Taken together, these studies suggest that secretory vesicle mobility is regulated by specific classes of secretagogues and that neuropeptide containing secretory vesicles may be released from dendritic structures.
Washburn
C L
CL
Center for Research on Occupational and Environmental Toxicology, The Vollum Institute Portland, OR 97201, USA.
Bean
J E
JE
Silverman
M A
MA
Pellegrino
M J
MJ
Yates
P A
PA
Allen
R G
RG
D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism; D001692:Biological Transport; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology; D007700:Kinetics; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009479:Neuropeptides / Q000187:drug effects / Q000378:metabolism*; D018847:Opioid Peptides / Q000235:genetics / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D022142:Secretory Vesicles / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D053379:Chromogranin A; D002864:Chromogranins; D008164:Luminescent Proteins; D009479:Neuropeptides; D018847:Opioid Peptides; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C444646:prepro-orphanin FQ (154-181); D049452:Green Fluorescent Proteins; D005576:Colforsin