TY - JOUR
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
AU - Secondo, Agnese
AU - Taglialatela, Maurizio
AU - Cataldi, Mauro
AU - Giorgio, Giovanna
AU - Valore, Monica
AU - Di Renzo, Gianfranco
AU - Annunziato, Lucio
PY - 2000
Y1 - 2000
N2 - 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+](i) oscillations 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+](i) achieved 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.
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+](i) oscillations 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+](i) achieved 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.
UR - http://www.scopus.com/inward/record.url?scp=0033790261&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033790261&partnerID=8YFLogxK
M3 - Article
C2 - 11040061
AN - SCOPUS:0033790261
VL - 58
SP - 1115
EP - 1128
JO - Molecular Pharmacology
JF - Molecular Pharmacology
SN - 0026-895X
IS - 5
ER -