title: Insulin/phosphoinositide 3-kinase pathway accelerates the glucose-induced first-phase insulin secretion through TrpV2 recruitment in pancreatic β-cells.
abstract: Functional insulin receptor and its downstream effector PI3K (phosphoinositide 3-kinase) have been identified in pancreatic β-cells, but their involvement in the regulation of insulin secretion from β-cells remains unclear. In the present study, we investigated the physiological role of insulin and PI3K in glucose-induced biphasic insulin exocytosis in primary cultured β-cells and insulinoma Min6 cells using total internal reflection fluorescent microscopy. The pretreatment of β-cells with insulin induced the rapid increase in intracellular Ca2+ levels and accelerated the exocytotic response without affecting the second-phase insulin secretion. The inhibition of PI3K not only abolished the insulin-induced rapid development of the exocytotic response, but also potentiated the second-phase insulin secretion. The rapid development of Ca2+ and accelerated exocytotic response induced by insulin were accompanied by the translocation of the Ca2+-permeable channel TrpV2 (transient receptor potential V2) in a PI3K-dependent manner. Inhibition of TrpV2 by the selective blocker tranilast, or the expression of shRNA (short-hairpin RNA) against TrpV2 suppressed the effect of insulin in the first phase, but the second phase was not affected. Thus our results demonstrate that insulin treatment induced the acceleration of the exocytotic response during the glucose-induced first-phase response by the insertion of TrpV2 into the plasma membrane in a PI3K-dependent manner.
authors.affiliation: Department of Biochemistry, Kyorin University School of Medicine, Mitaka, Tokyo 181-8611, Japan.
authors.forename: Kyota
authors.initials: K
authors.lastname: Aoyagi
authors.forename: Mica
authors.initials: M
authors.lastname: Ohara-Imaizumi
authors.forename: Chiyono
authors.initials: C
authors.lastname: Nishiwaki
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nakamichi
authors.forename: Shinya
authors.initials: S
authors.lastname: Nagamatsu
mesh_terms: D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000235:genetics*; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005089:Exocytosis; D013006:Growth Hormone / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000502:physiology*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D050916:TRPV Cation Channels / Q000235:genetics*; D014162:Transfection
chemical_list: D015220:Calcium Channels; D018076:DNA, Complementary; D007328:Insulin; D050916:TRPV Cation Channels; C485717:Trpv2 protein, mouse; D013006:Growth Hormone; D004247:DNA; D019869:Phosphatidylinositol 3-Kinases