Imidazoline NNC77-0074 stimulates insulin secretion and inhibits glucagon release by control of Ca(2+)-dependent exocytosis in pancreatic alpha- and beta-cells

Eur J Pharmacol. 2003 Apr 11;466(1-2):213-21. doi: 10.1016/s0014-2999(03)01537-1.

Abstract

We have investigated the effects of the novel imidazoline compound (+)-2-(2-(4,5-dihydro-1H-imidazol-2-yl)-thiopene-2-yl-ethyl)-pyridine (NNC77-0074) on stimulus-secretion coupling in isolated pancreatic alpha- and beta-cells. NNC77-0074 stimulated glucose-dependent insulin secretion in intact mouse pancreatic islets. No effect was observed at </=2.5 mM glucose and maximal stimulation occurred at 10-15 mM glucose. NNC77-0074 produced a concentration-dependent stimulation of insulin secretion. Half-maximal (EC(50)) stimulation was observed at 24 microM and at maximally stimulatory concentrations insulin release was doubled. The stimulatory action of NNC77-0074 on insulin secretion was not associated with membrane depolarisation or a change in the activity of ATP-sensitive K(+) channels. Using capacitance measurements, we found that NNC77-0074 stimulated depolarisation-induced exocytosis 2.6-fold without affecting the whole-cell Ca(2+) current when applied via the extracellular medium. The concentration dependence of the stimulatory action was determined by intracellular application of NNC77-0074 through the recording pipette. NNC77-0074 stimulated exocytosis half-maximal at 44 nM and at maximally stimulatory concentrations the rate of exocytosis was increased twofold. NNC77-0074 stimulated depolarised-induced insulin secretion from islets exposed to diazoxide and high external KCl (EC(50)=0.45 microM). The stimulatory action of NNC77-0074 was dependent on protein kinase C activity. NNC77-0074 potently inhibited glucagon secretion from rat islets (EC(50)=11 nM). This was not associated with a change in spontaneous electrical activity and ATP-sensitive K(+) channel activity but resulted from a reduction of the rate of Ca(2+)-dependent exocytosis in single rat alpha-cells (EC(50)=9 nM). Inhibition of exocytosis by NNC77-0074 was pertussis toxin-sensitive and mediated by activation of the protein phosphatase calcineurin. In rat somatotrophs, PC12 cells and mouse cortical neurons NNC77-0074 did not stimulate Ca(2+)-evoked exocytosis, whereas the other imidazoline compounds phentolamine and efaroxan produced 2.5-fold stimulation of exocytosis. Our data suggest that the imidazoline compound NNC77-0074 constitutes a novel class of antidiabetic compounds that stimulates glucose-dependent insulin release while inhibiting glucagon secretion. These actions are exclusively exerted by modulation of exocytosis of the insulin- and glucagon-containing granules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Calcium Channels / physiology
  • Cells, Cultured
  • Cyclic AMP / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Exocytosis
  • Female
  • Glucagon / antagonists & inhibitors
  • Glucagon / metabolism*
  • Glucose / metabolism
  • Imidazoles / pharmacology*
  • In Vitro Techniques
  • Insulin / metabolism*
  • Insulin Secretion
  • Islets of Langerhans / drug effects*
  • Islets of Langerhans / enzymology
  • Islets of Langerhans / physiology
  • Mice
  • Mice, Inbred Strains
  • Patch-Clamp Techniques
  • Potassium Channels / physiology
  • Protein Kinase C / physiology
  • Pyridines / pharmacology*

Substances

  • 2-(2-(4,5-dihydro-1H-imidazol-2-yl)thiophene-2-ylethyl)pyridine
  • Calcium Channels
  • Enzyme Inhibitors
  • Imidazoles
  • Insulin
  • Potassium Channels
  • Pyridines
  • Glucagon
  • Cyclic AMP
  • Protein Kinase C
  • Glucose
  • Calcium