The effects of cromakalim on ATP-sensitive potassium channels in insulin-secreting cells.
1. The single-channel current recording technique has been used to investigate the effects of cromakalim, diazoxide and ATP, separately and combined, on the opening of ATP-sensitive potassium channels in the insulin-secreting cell-line RINm5F. The actions of these drugs have been studied using the permeabilized open-cell variation of the patch-clamp technique. 2. In the absence of internal ATP, cromakalim (80-200 microM) was unable to open ATP-sensitive K+ channels but when ATP was present both cromakalim and diazoxide caused channel openings. 3. Interactions between ATP and cromakalim seemed competitive. Concentrations of cromakalim in the range 80-200 microM readily activated channels inhibited by 0.1 mM ATP, but had no effects when the concentration of ATP was increased to 0.5-2 mM. Only when the concentration of cromakalim was increased to 400-800 microM could opening of 0.5-2 mM ATP-inhibited channels be regularly observed. In the continued presence of cromakalim (400-800 microM), an increase in the internal concentration of ATP from either 0.25 to 0.5 mM or 1 to 2 mM, inhibited cromakalim-activated K+ channels. 4. Activation of ATP-inhibited K+ channels was abolished by replacing ATP with ATP gamma S and cromakalin had no effects on ATP gamma S-inhibited channels. This suggests that cromakalim may open KATP channels in insulin-secreting cells by a mechanism which involves protein phosphorylation.
Dunne
M J
MJ
Physiological Laboratory, University of Liverpool.
Aspinall
R J
RJ
Petersen
O H
OH
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D001578:Benzopyrans / Q000494:pharmacology*; D002460:Cell Line; D019806:Cromakalim; D003981:Diazoxide / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015221:Potassium Channels / Q000187:drug effects*; D011197:Potassium Radioisotopes; D011758:Pyrroles / Q000494:pharmacology*; D012414:Rubidium Radioisotopes; D012964:Sodium / Q000502:physiology
D001578:Benzopyrans; D007328:Insulin; D015221:Potassium Channels; D011197:Potassium Radioisotopes; D011758:Pyrroles; D012414:Rubidium Radioisotopes; D019806:Cromakalim; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D012964:Sodium; D003981:Diazoxide