title: Membrane potential dependent modulations of calcium oscillations in insulin-secreting INS-1 cells.
abstract: This study was undertaken to examine the role of K(+) channels on cytosolic Ca(2+) ([Ca(2+)](i)) in insulin secreting cells. [Ca(2+)](i) was measured in single glucose-responsive INS-1 cells using the fluorescent Ca(2+) indicator Fura-2. Glucose, tolbutamide and forskolin elevated [Ca(2+)](i) and induced [Ca(2+)] oscillations. Whereas the glucose effect was delayed and observed in 60% and 93% of the cells, in a poorly and a highly glucose-responsive INS-1 cell clone, respectively, tolbutamide and forskolin increased [Ca(2+)](i) in all cells tested. In the latter clone, glucose induced [Ca(2+)](i) oscillations in 77% of the cells. In 16% of the cells a sustained rise of [Ca(2+)](i) was observed. The increase in [Ca(2+)](i) was reversed by verapamil, an L-type Ca(2+) channel inhibitor. Adrenaline decreased [Ca(2+)](i) in oscillating cells in the presence of low glucose and in cells stimulated by glucose alone or in combination with tolbutamide and forskolin. Adrenaline did not lower [Ca(2+)](i) in the presence of 30mM extracellular K(+), indicating that adrenaline does not exert a direct effect on Ca(2+) channels but increases K(+) channel activity. As for primary b-cells, [Ca(2+)](i) oscillations persisted in the presence of closed K(ATP) channels; these also persisted in the presence of thapsigargin, which blocks Ca(2+) uptake into Ca(2+) stores. In contrast, in voltage-clamped cells and in the presence of diazoxide (50mM), which hyperpolarizes the cells by opening K(ATP) channels, [Ca(2+)](i) oscillations were abolished. These results support the hypothesis that [Ca(2+)](i) oscillations depend on functional voltage-dependent Ca(2+) and K(+) channels and are interrupted by a hyperpolarization in insulin-secreting cells.
authors.affiliation: Physiologisches Institut II, Universität Freiburg, Freiburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Herbst
authors.forename: P
authors.initials: P
authors.lastname: Sasse
authors.forename: R
authors.initials: R
authors.lastname: Greger
authors.forename: H
authors.initials: H
authors.lastname: Yu
authors.forename: J
authors.initials: J
authors.lastname: Hescheler
authors.forename: S
authors.initials: S
authors.lastname: Ullrich
mesh_terms: D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002460:Cell Line; D003000:Clonidine / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D003981:Diazoxide / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004837:Epinephrine / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D016257:Fura-2 / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology*; D051379:Mice; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000378:metabolism*; D011224:Prazosin / Q000494:pharmacology; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology; D014044:Tolbutamide / Q000494:pharmacology
chemical_list: D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007004:Hypoglycemic Agents; D007328:Insulin; D015221:Potassium Channels; D018341:Receptors, Adrenergic, alpha-2; D005576:Colforsin; D019284:Thapsigargin; D014044:Tolbutamide; D005947:Glucose; D003000:Clonidine; D003981:Diazoxide; D002118:Calcium; D016257:Fura-2; D011224:Prazosin; D004837:Epinephrine