Functional VIP receptors in the human mucus-secreting colonic epithelial cell line CL.16E.
A polarized human clonal intestinal cell line exhibiting mucus secretion (Cl.16E) was used to study the expression and function of vasoactive intestinal peptide (VIP) receptors in mucus-secreting cells. Cl.16E cells expressed one class of receptors with a KD of 0.13 nM and a capacity of 67 fmol/mg protein. Covalent labeling of receptors followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a receptor protein with a Mr of 63,000 in Cl.16E cells. VIP stimulated adenylate cyclase activity in membranes from Cl.16E cells with an ED50 of 0.06 nM. In conditions where carbachol stimulated mucin secretion from filter-grown Cl.16E cells, VIP, dibutyryl adenosine 3',5'-cyclic monophosphate (DbcAMP), or forskolin did not alter basal secretion. However, VIP strongly potentiated carbachol-induced mucin secretion, since carbachol alone and VIP plus carbachol induced a 1.6- and 3.6-fold increase of mucin secretion above basal, respectively. This potentiating effect of VIP was mimicked by DbcAMP or forskolin. It was observed for VIP concentrations in the 0.1-100 nM range (ED50, 2 nM). VIP elicited a dramatic increase of intracellular cAMP levels in filter-grown Cl.16E cells with a dose-response curve (ED50, 4 nM) very similar to that observed for the modulation of mucin secretion. These studies suggest that the clonal cell line Cl.16E may be an invaluable cellular model for evaluating the neurohormonal control of mucus secretion.
Laburthe
M
M
Unité de Recherche sur la Différenciation et la Neuroendocrinologie de Cellules Digestives, INSERM U178, Villejuif, France.
Augeron
C
C
Rouyer-Fessard
C
C
Roumagnac
I
I
Maoret
J J
JJ
Grasset
E
E
Laboisse
C
C
D003994:Bucladesine / Q000494:pharmacology; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D003106:Colon; D004848:Epithelium; D006801:Humans; D007700:Kinetics; D009077:Mucins / Q000378:metabolism; D009093:Mucus / Q000187:drug effects / Q000378:metabolism*; D011964:Receptors, Gastrointestinal Hormone / Q000378:metabolism*; D018005:Receptors, Vasoactive Intestinal Peptide; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism* / Q000494:pharmacology
D009077:Mucins; D011964:Receptors, Gastrointestinal Hormone; D018005:Receptors, Vasoactive Intestinal Peptide; D005576:Colforsin; D014660:Vasoactive Intestinal Peptide; D003994:Bucladesine; D002217:Carbachol