Sensitivity to dihydropyridines, omega-conotoxin and noradrenaline reveals multiple high-voltage-activated Ca2+ channels in rat insulinoma and human pancreatic beta-cells.
High-voltage-activated (HVA) Ba2+ currents of rat insulinoma (RINm5F) and human pancreatic beta-cells were tested for their sensitivity to dihydropyridines (DHPs), omega-conotoxin (omega-CgTx) and noradrenaline. In RINm5F cells, block of HVA currents by nimodipine, nitrendipine and nifedipine was voltage- and dose-dependent (apparent KD < 37 nM) and largely incomplete even at saturating doses of DHPs (mean 53%, at 10 microM and 0 mV). Analysis of slow tail currents in Bay K 8644-treated cells indicated the existence of Bay K 8644-insensitive channels that turned on at slightly more positive voltages and deactivated more quickly than Bay K 8644-modified channels. DHP Ca2+ agonists and antagonists in human beta-cells had similar features to RINm5F cells except that DHP block was more pronounced (76%, at 10 microM and 0 mV) and Bay K 8644 action was more effective, suggesting a higher density of L-type Ca2+ channels in these cells. In RINm5F cells, but not in human beta-cells, DHP-resistant currents were sensitive to omega-CgTx. The toxin depressed 10-20% of the DHP-resistant currents sparing a "residual" current (25-35%) with similar voltage-dependent characteristics and Ca2+/Ba2+ permeability. Noradrenaline (10 microM) exhibited different actions on the various HVA current components: (1) it prolonged the activation kinetics of omega-CgTx-sensitive currents, (2) it depressed by about 20% the size of DHP-sensitive currents, and (3) it had little or no effects on the residual DHP- and omega-CgTx-resistant current although intracellularly applied guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) prolonged its activation time course.(ABSTRACT TRUNCATED AT 250 WORDS)
Pollo
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A
Department of Anatomy and Human Physiology, Corso Raffaello 30, Turin, Italy.
Lovallo
M
M
Biancardi
E
E
Sher
E
E
Socci
C
C
Carbone
E
E
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D001464:Barium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D015220:Calcium Channels / Q000187:drug effects*; D004095:Dihydropyridines / Q000494:pharmacology; D004594:Electrophysiology; D006801:Humans; D007340:Insulinoma / Q000378:metabolism*; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D009638:Norepinephrine / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D051381:Rats; D014407:Tumor Cells, Cultured; D020866:omega-Conotoxin GVIA
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D004095:Dihydropyridines; D010455:Peptides; D001464:Barium; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D020866:omega-Conotoxin GVIA; D009638:Norepinephrine