title: TRPM4 controls insulin secretion in pancreatic beta-cells.
abstract: TRPM4 is a calcium-activated non-selective cation channel that is widely expressed and proposed to be involved in cell depolarization. In excitable cells, TRPM4 may regulate calcium influx by causing the depolarization that drives the activation of voltage-dependent calcium channels. We here report that insulin-secreting cells of the rat pancreatic beta-cell line INS-1 natively express TRPM4 proteins and generate large depolarizing membrane currents in response to increased intracellular calcium. These currents exhibit the characteristics of TRPM4 and can be suppressed by expressing a dominant negative TRPM4 construct, resulting in significantly decreased insulin secretion in response to a glucose stimulus. Reduced insulin secretion was also observed with arginine vasopressin stimulation, a Gq-coupled receptor agonist in beta-cells. Moreover, the recruitment of TRPM4 currents was biphasic in both INS-1 cells as well as HEK-293 cells overexpressing TRPM4. The first phase is due to activation of TRPM4 channels localized within the plasma membrane followed by a slower secondary phase, which is caused by the recruitment of TRPM4-containing vesicles to the plasma membrane during exocytosis. The secondary phase can be observed during perfusion of cells with increasing [Ca(2+)](i), replicated with agonist stimulation, and coincides with an increase in cell capacitance, loss of FM1-43 dye, and vesicle fusion. Our data suggest that TRPM4 may play a key role in the control of membrane potential and electrical activity of electrically excitable secretory cells and the dynamic translocation of TRPM4 from a vesicular pool to the plasma membrane via Ca(2+)-dependent exocytosis may represent a key short- and midterm regulatory mechanism by which cells regulate electrical activity.
authors.affiliation: Laboratory of Cell and Molecular Signaling, Center for Biomedical Research at The Queen's Medical Center and John A. Burns School of Medicine at the University of Hawaii, 1301 Punchbowl Street, UHT 8, Honolulu, HI 96813, USA.
authors.forename: Henrique
authors.initials: H
authors.lastname: Cheng
authors.forename: Andreas
authors.initials: A
authors.lastname: Beck
authors.forename: Pierre
authors.initials: P
authors.lastname: Launay
authors.forename: Stefan A
authors.initials: SA
authors.lastname: Gross
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Stokes
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Kinet
authors.forename: Andrea
authors.initials: A
authors.lastname: Fleig
authors.forename: Reinhold
authors.initials: R
authors.lastname: Penner
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D002460:Cell Line; D005089:Exocytosis; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism*; D008564:Membrane Potentials; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D050053:TRPM Cation Channels / Q000235:genetics / Q000378:metabolism*
chemical_list: D007328:Insulin; D011994:Recombinant Proteins; D050053:TRPM Cation Channels; C500239:TRPM4 protein, rat; D005947:Glucose