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. 1994 Apr 15;299(Pt 2):579–585. doi: 10.1042/bj2990579

Role of calcium in carbachol- and neurotensin-induced mucin exocytosis in a human colonic goblet cell line and cross-talk with the cyclic AMP pathway.

C Bou-Hanna 1, B Berthon 1, L Combettes 1, M Claret 1, C L Laboisse 1
PMCID: PMC1138310  PMID: 8172620

Abstract

The mechanisms of Ca(2+)-induced mucin secretion were examined in monolayers of the differentiated epithelial colon cell line C1.16E by combined measurements of free intracellular Ca2+ ([Ca2+]i) using a fluorescence indicator and mucous secretion using a specific and sensitive electrophoretic assay. Carbachol, a cholinergic agonist, induced an initial concentration-dependent [Ca2+]i peak increasing from 129 +/- 3 nM (basal [Ca2+]i) to 608 +/- 101 nM at 1 x 10(-4) M carbachol with an ED50 of 7 microM, and this was followed by a lower-level plateau. These biphasic effects were reversed by the muscarinic-receptor antagonist atropine. In the absence of extracellular Ca2+, the initial [Ca2+]i peak was maintained while the sustained plateau was abolished. The regulatory peptide neurotensin caused a monophasic transient rise in [Ca2+]i followed by a very rapid return to baseline. The neurotensin-induced rise in [Ca2+]i was concentration-dependent with an ED50 of 4 nM, and was maximal at 1 x 10(-6) M (598 +/- 127 nM). The [Ca2+]i response to neurotensin was not significantly affected by extracellular Ca2+ depletion. Carbachol-induced mucin exocytosis was concentration-dependent with an ED50 of 15 microM, and was inhibited by 35% upon removal of extracellular Ca2+. Neurotensin caused a concentration-dependent rise in mucous secretion with an ED50 of 36 nM, not significantly affected upon removal of extracellular Ca2+. Together our results suggest that while the mucin secretory response to carbachol depends on both the release of Ca2+ from intracellular stores and a Ca2+ influx from external medium, the secretory response to neurotensin is based solely on intracellular Ca2+ mobilization. Finally, evaluation of the cross-talk between the cyclic AMP pathway stimulated by vasoactive intestinal peptide (VIP) and the Ca2+ pathway stimulated by neurotensin or carbachol led to the conclusion that the potentiated secretory response elicited by the combined action of carbachol and VIP requires extracellular Ca2+.

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Selected References

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