Expression, processing, and secretion of the neuroendocrine VGF peptides by INS-1 cells.
The neurotropin-inducible gene vgf is expressed in neuronal and endocrine tissues. It encodes a secretory protein that is proteolytically processed in neuronal cells to low molecular mass polypeptides. In the present report, we show that vgf is expressed in different insulinoma cell lines and in normal rat pancreatic islets. In the insulinoma-derived beta-cell line INS-1, vgf messenger RNA was transcriptionally up-regulated by increased levels ofintracellular cAMP, but not by the addition of glucose (20 mM) or phorbol 12-myristate 13-acetate (100 nM). Furthermore, nerve growth factor failed to stimulate vgf gene expression. In INS-1 cells, the VGF protein was shown to be processed in a post endoplasmic reticulum compartment to produce a peptide profile similar to that seen in neurons. The release of such VGF peptides occurred at a low rate in the absence of secretory stimuli (<2%/h). A 3-fold increase in the rate of release was seen after the addition of glucose (15 mM), a 4-fold increase was seen after (Bu)2cAMP (1 mM), and a 6-fold increase was seen after phorbol 12-myristate 13-acetate (100 nM). These results indicated that insulin-containing cells produce VGF-derived peptides that are released via a regulated pathway in response to insulin secretagogues.
Possenti
R
R
Department of Neuroscience, University of Tor Vergata, Rome, Italy. r.possenti@in.rm.cnr.it
Rinaldi
A M
AM
Ferri
G L
GL
Borboni
P
P
Trani
E
E
Levi
A
A
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D000242:Cyclic AMP / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007340:Insulinoma; D007700:Kinetics; D009479:Neuropeptides; D016716:PC12 Cells; D010190:Pancreatic Neoplasms; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured
D007328:Insulin; D009479:Neuropeptides; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C062040:Vgf protein, rat; D003994:Bucladesine; D000242:Cyclic AMP; D015500:Chloramphenicol O-Acetyltransferase; D005947:Glucose; D013755:Tetradecanoylphorbol Acetate