title: Melatonin influences insulin secretion primarily via MT(1)  receptors in rat insulinoma cells (INS-1) and mouse pancreatic islets.
abstract: Several studies have revealed that melatonin affects the insulin secretion via MT(1)  and MT(2)  receptor isoforms. Owing to the lack of selective MT(1)  receptor antagonists, we used RNA interference technology to generate an MT(1)  knockdown in a clonal β-cell line to evaluate whether melatonin modulates insulin secretion specifically via the MT(1)  receptor. Incubation experiments were carried out, and the insulin concentration in supernatants was measured using a radioimmunoassay. Furthermore, the intracellular cAMP was determined using an enzyme-linked immunosorbent assay. Real-time RT-PCR indicated that MT(1)  knockdown resulted in a significant increase in the rIns1 mRNA and a significantly elevated basal insulin secretion of INS-1 cells. Incubation with melatonin decreased the amount of glucagon-like peptide 1 or inhibited the glucagon-stimulated insulin release of INS-1 cells, while, in MT(1) -knockdown cells, no melatonin-induced reduction in insulin secretion could be found. No decrease in 3-isobutyl-1-methylxanthine-stimulated intracellular cAMP in rMT(1) -knockdown cells was detectable after treatment with melatonin either, and immunocytochemistry proved that MT(1)  knockdown abolished phosphorylation of cAMP-response-element-binding protein. In contrast to the INS-1 cells, preincubation with melatonin did not sensitize the insulin secretion of rMT(1) -knockdown cells. We also monitored insulin secretion from isolated islets of wild-type and melatonin-receptor knockout mice ex vivo. In islets of wild-type mice, melatonin treatment resulted in a decrease in insulin release, whereas melatonin treatment of islets from MT(1)  knockout and MT(1/2)  double-knockout mice did not show a significant effect. The data indicate that melatonin inhibits insulin secretion, primarily via the MT(1)  receptor in rat INS-1 cells and isolated mouse islets.
authors.affiliation: Saxon Academy of Sciences Leipzig, Leipzig, Germany. eckhard.muehlbauer@medizin.unihalle.de
authors.forename: Eckhard
authors.initials: E
authors.lastname: Mühlbauer
authors.forename: Elke
authors.initials: E
authors.lastname: Albrecht
authors.forename: Ivonne
authors.initials: I
authors.lastname: Bazwinsky-Wutschke
authors.forename: Elmar
authors.initials: E
authors.lastname: Peschke
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D045744:Cell Line, Tumor; D000242:Cyclic AMP / Q000378:metabolism; D006651:Histocytochemistry; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D007340:Insulinoma / Q000378:metabolism; D008550:Melatonin / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D051381:Rats; D044122:Receptor, Melatonin, MT1 / Q000235:genetics / Q000378:metabolism*; D018709:Statistics, Nonparametric
chemical_list: D007328:Insulin; D044122:Receptor, Melatonin, MT1; D000242:Cyclic AMP; D008550:Melatonin; D015056:1-Methyl-3-isobutylxanthine