Regulation of neurotensin release from canine enteric primary cell cultures.
A recently developed primary cell-culture system allows direct study of the cellular mechanisms regulating neurotensin secretion from intestinal mucosal cells. We now report the use of these methods to evaluate the modulation of neurotensin release by adrenergic, cholinergic, and peptidergic transmitters. Collagenase-dispersed canine ileal mucosal cells, enriched for neurotensinlike immunoreactivity (NTLI) by centrifugal elutriation, were maintained for 48 h on collagen-coated culture dishes. Epinephrine (0.01-100 microM) stimulated a dose-dependent increase increase in NTLI secretion. The NTLI response to epinephrine increase in NTLI secretion. The NTLI response to epinephrine was competitively inhibited by propranolol, producing a parallel rightward shift of the epinephrine dose-response curve. alpha-Adrenergic agonist methoxamine (10 microM) and clonidine (10 microM) did not alter basal NTLI secretion. Epinephrine stimulation was not significantly inhibited by the alpha-adrenergic antagonists prazosin (10 microM) or yohimbine (10 microM). The diterpene forskolin, an adenyl cyclase activator, increased NTLI release and had an additive effect on the response to epinephrine. In contrast to beta-adrenergic activation, carbachol and somatostatin produced a dose-dependent inhibition of epinephrine-stimulated NTLI release. At 100 microM carbachol, NTLI release was inhibited 68%, and this action was partially blocked by atropine (0.1 microM). Somatostatin (100 nM) produced a 96% inhibition that was not surmountable by 1 mM epinephrine. These data indicate that neurotensin release is stimulated by beta-adrenergic agonists and adenylate cyclase activation. Somatostatin and the muscarinic agonist carbachol directly inhibit NTLI release.
Barber
D L
DL
Buchan
A M
AM
Walsh
J H
JH
Soll
A H
AH
D000262:Adenylyl Cyclases / Q000378:metabolism; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D004285:Dogs; D004837:Epinephrine / Q000494:pharmacology; D007082:Ileum / Q000166:cytology / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009479:Neuropeptides; D009496:Neurotensin / Q000378:metabolism*; D010277:Parasympathomimetics / Q000494:pharmacology; D011863:Radioimmunoassay; D013004:Somatostatin / Q000494:pharmacology; D013997:Time Factors
D000318:Adrenergic beta-Agonists; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D010277:Parasympathomimetics; C031644:neurotensin-like immunoreactivity; D005576:Colforsin; D009496:Neurotensin; D013004:Somatostatin; D002217:Carbachol; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D004837:Epinephrine