RT Journal Article
SR Electronic
T1 Pharmacological Blockade of ERG K+ Channels and Ca2+ Influx through Store-Operated Channels Exerts Opposite Effects on Intracellular Ca2+ Oscillations in Pituitary GH3 Cells
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP 1115
OP 1128
DO 10.1124/mol.58.5.1115
VO 58
IS 5
A1 Agnese Secondo
A1 Maurizio Taglialatela
A1 Mauro Cataldi
A1 Giovanna Giorgio
A1 Monica Valore
A1 Gianfranco Di Renzo
A1 Lucio Annunziato
YR 2000
UL http://molpharm.aspetjournals.org/content/58/5/1115.abstract
AB In the present study, the effects on intracellular calcium concentration ([Ca2+]i) oscillations of the blockade of ether-a-go-go-related gene (ERG) K+channels and of Ca2+ influx through store-operated channels (SOC) activated by [Ca2+]i store depletion have been studied in GH3 cells by means of a combination of single-cell fura-2 microfluorimetry and whole-cell mode of the patch-clamp technique. Nanomolar concentrations (1–30 nM) of the piperidinic second-generation antihistamines terfenadine and astemizole and of the class III antiarrhythmic methanesulfonanilide dofetilide, by blocking ERG K+ channels, increased the frequency and the amplitude of [Ca2+]i oscillations in resting oscillating GH3 cells. These compounds also induced the appearance of an oscillatory pattern of [Ca2+]i in a subpopulation of nonoscillating GH3 cells. The effects of ERG K+ channel blockade on [Ca2+]i oscillations appeared to be due to the activation of L-type Ca2+ channels, because they were prevented by 300 nM nimodipine. By contrast, the piperazinic second-generation antihistamine cetirizine (0.01–30 μM), which served as a negative control, failed to affect ERG K+channels and did not interfere with [Ca2+]ioscillations in GH3 cells. Interestingly, micromolar concentrations of terfenadine and astemizole (0.3–30 μM), but not of dofetilide (10–100 μM), produced an inhibition of the spontaneous oscillatory pattern of [Ca2+]i changes. This effect was possibly related to an inhibition of SOC, because these compounds inhibited the increase of [Ca2+]iachieved by extracellular calcium reintroduction after intracellular calcium store depletion with the sarcoplasmic or endoplasmic reticulum calcium ATPase pump inhibitor thapsigargin (10 μM) in an extracellular calcium-free medium. The same inhibitory effect on [Ca2+]i oscillations and SOC was observed with the first-generation antihistamine hydroxyzine (1–30 μM), the more hydrophobic metabolic precursor of cetirizine. Collectively, the results of the present study obtained with compounds that interfere in a different concentration range with ERG K+ channels or SOC suggest that 1) ERG K+ channels play a relevant role in controlling the oscillatory pattern of [Ca2+]i in resting GH3 cells and 2) the inhibition of SOC might induce an opposite effect, i.e., an inhibition of [Ca2+]i oscillations.