Characterization of ATP-sensitive potassium channels functionally expressed in pituitary GH3 cells

Sheng-Nan Wu, H. F. Li, H. T. Chiang

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

ATP-sensitive K+ (K(ATP)) channels have been characterized in pituitary GH3 cells with the aid of the patch-clamp technique. In the cell-attached configuration, the presence of diazoxide (100 μM) revealed the presence of glibenclamide-sensitive K(ATP) channel exhibiting a unitary conductance of 74 pS. Metabolic inhibition induced by 2,4-dinitrophenol (1 mM) or sodium cyanide (300 μM) increased K(ATP) channel activity, while nicorandil (100 μM) had no effect on it. In the inside-out configuration, Mg-ATP applied intracellularly suppressed the activity of K(ATP) channels in a concentration-dependent manner with an IC50 value of 30 μM. The activation of phospholipase A2 caused by mellitin (1 μM) was found to enhance K(ATP) channel activity and further application of aristolochic acid (30 μM) reduced the mellitin-induced increase in channel activity. The challenging of cells with 4,4'-dithiodipyridine (100 μM) also induced K(ATP) channel activity. Diazoxide, mellitin and 4,4'-dithiodipyridine activated the K(ATP) channels that exhibited similar channel-opening kinetics. In addition, under current-clamp conditions, the application of diazoxide (100 μM) hyperpolarized the membrane potential and reduced the firing rate of spontaneous action potentials. The present study clearly indicates that K(ATP) channels similar to those seen in pancreatic β cells are functionally expressed in GH3 cells. In addition to the presence of Ca2+-activated K+ channels, K(ATP) channels found in these cells could thus play an important role in controlling hormonal release by regulating the membrane potential.

Original languageEnglish
Pages (from-to)205-214
Number of pages10
JournalJournal of Membrane Biology
Volume178
Issue number3
DOIs
Publication statusPublished - 2000 Dec 1

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KATP Channels
Adenosine Triphosphate
Melitten
Diazoxide
Membrane Potentials
Sodium Cyanide
Nicorandil
2,4-Dinitrophenol
Calcium-Activated Potassium Channels
Glyburide
Phospholipases A2
Patch-Clamp Techniques
Action Potentials
Inhibitory Concentration 50

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Physiology
  • Cell Biology

Cite this

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abstract = "ATP-sensitive K+ (K(ATP)) channels have been characterized in pituitary GH3 cells with the aid of the patch-clamp technique. In the cell-attached configuration, the presence of diazoxide (100 μM) revealed the presence of glibenclamide-sensitive K(ATP) channel exhibiting a unitary conductance of 74 pS. Metabolic inhibition induced by 2,4-dinitrophenol (1 mM) or sodium cyanide (300 μM) increased K(ATP) channel activity, while nicorandil (100 μM) had no effect on it. In the inside-out configuration, Mg-ATP applied intracellularly suppressed the activity of K(ATP) channels in a concentration-dependent manner with an IC50 value of 30 μM. The activation of phospholipase A2 caused by mellitin (1 μM) was found to enhance K(ATP) channel activity and further application of aristolochic acid (30 μM) reduced the mellitin-induced increase in channel activity. The challenging of cells with 4,4'-dithiodipyridine (100 μM) also induced K(ATP) channel activity. Diazoxide, mellitin and 4,4'-dithiodipyridine activated the K(ATP) channels that exhibited similar channel-opening kinetics. In addition, under current-clamp conditions, the application of diazoxide (100 μM) hyperpolarized the membrane potential and reduced the firing rate of spontaneous action potentials. The present study clearly indicates that K(ATP) channels similar to those seen in pancreatic β cells are functionally expressed in GH3 cells. In addition to the presence of Ca2+-activated K+ channels, K(ATP) channels found in these cells could thus play an important role in controlling hormonal release by regulating the membrane potential.",
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Characterization of ATP-sensitive potassium channels functionally expressed in pituitary GH3 cells. / Wu, Sheng-Nan; Li, H. F.; Chiang, H. T.

In: Journal of Membrane Biology, Vol. 178, No. 3, 01.12.2000, p. 205-214.

Research output: Contribution to journalArticle

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AB - ATP-sensitive K+ (K(ATP)) channels have been characterized in pituitary GH3 cells with the aid of the patch-clamp technique. In the cell-attached configuration, the presence of diazoxide (100 μM) revealed the presence of glibenclamide-sensitive K(ATP) channel exhibiting a unitary conductance of 74 pS. Metabolic inhibition induced by 2,4-dinitrophenol (1 mM) or sodium cyanide (300 μM) increased K(ATP) channel activity, while nicorandil (100 μM) had no effect on it. In the inside-out configuration, Mg-ATP applied intracellularly suppressed the activity of K(ATP) channels in a concentration-dependent manner with an IC50 value of 30 μM. The activation of phospholipase A2 caused by mellitin (1 μM) was found to enhance K(ATP) channel activity and further application of aristolochic acid (30 μM) reduced the mellitin-induced increase in channel activity. The challenging of cells with 4,4'-dithiodipyridine (100 μM) also induced K(ATP) channel activity. Diazoxide, mellitin and 4,4'-dithiodipyridine activated the K(ATP) channels that exhibited similar channel-opening kinetics. In addition, under current-clamp conditions, the application of diazoxide (100 μM) hyperpolarized the membrane potential and reduced the firing rate of spontaneous action potentials. The present study clearly indicates that K(ATP) channels similar to those seen in pancreatic β cells are functionally expressed in GH3 cells. In addition to the presence of Ca2+-activated K+ channels, K(ATP) channels found in these cells could thus play an important role in controlling hormonal release by regulating the membrane potential.

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